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A platform to address questions regarding methods and development of new methods.
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Hi everyone,
has anyone used Zeba spin desalting columns?
Can they be reused?
They are quite expensive and it would be great if they can be reused...
Also, manufacturer (Thermo Scientific) hasn't provided information whether they are polypropylene/polyethilene/dextran/sephadex or whatsoever-based columns...
Cheers,
Zeljko
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I would like to know the answer to this question as well. I use Zeba columns to buffer exchange proteins. I have tried double using before and I got my protein through the 2nd time just fine without loss, but I didn't have anyway to confirm if I actually exchanged the buffer.
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I am planning a citizen science project where participants will be asked to log the occurrence and magnitude of gastrointestinal events throughout the day. Is there an app that would be well suited for this (for both iOS and Android)? Ideally one that requires only the press of a button to log an event to keep it as simple as possible.
Thank you for any answers
Jonas
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Results from qualitative studies cannot be generalized. Therefore, is it possible to propose a social practice model based on the findings that cannot be generalized? I see some published articles and thesis that present models as a result of qualitative research. I will be glad if you help, thanks in advance :))
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According to qualitative paradigm, grounded theory is used to build a theory from a very big material such as more than 40 interviews for example. That's the purpose of inductive reasoning. Then, as Mohamed Salaheldeen said, you may test it by adopting a "deductive" method, namely building quantitative designs.
You certainly will find more details here :
Green J, Thorogood N. Qualitative Methods for Health Research. 2e éd. Londres: SAGE Publications Ltd; 2009. 304 p. (Introducing Qualitative Methods Series).
Regards,
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hi guyz!
i need your help..
im working now with my thesis..
i need some supplementary articles...unfortunately, i cant access here in our country articles from science direct....if you dont mind, kindly download this article for me...
doi:10.1016/j.phytochem.2004.04.013
thank you!
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This is this the main article.I did not see any supplementary materials
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Could anybody recommend a good method to isolate total RNA from human blood plasma?
We need to analyze some important patient samples. But we do not have whole blood - the technician only froze blood plasma! So I'm asking out of desperation.
We need to extract total RNA (we are particularly interested in mRNA). We have between 1 ml and 1.5 ml of plasma, according to the sample.
I am presuming that some cells might remain in the plasma after centrifugation (3500 rpm, 10 minutes) and its separation from the whole blood sample. Does anybody know approx. how many cells might be contaminating the plasma and what yield could be expected? Has anybody successfully extracted RNA this way before?
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Dear Ruth,
do Trizol and you will get whatever is in the plasma. However, mRNA will not be much but you certainly get miRNA.
Best
Boris
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In your opinion, what is the difference between a good teacher and a great teacher?
and what are the main teaching styles and how they Impact Students?
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Teaching is a 'calling' not a job. Hence a great teacher sees it as their 'calling'.
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Solutions
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Dear researcher, thanks a lot for nice question. Testosterone dissolves in chloroform at 100 mg/ml to yield a clear, colorless to very faint yellow solution. It is also soluble in methanol, absolute ethanol (0.5 mg/ml), 45 percent (w/v) aqueous 2-hydroxypropyl-b-cyclodextrin (18.2 mg/ml), vegetable oils and dioxane. https://doi.org/10.1038/s41598-020-60657-4
17β-Estradiol is soluble in organic solvents such as ethanol, DMSO, and dimethyl formamide (DMF). The solubility of estradiol in these solvents is at least 2.5 mg/ml in ethanol and 20 mg/ml in DMSO and DMF. Source: https://www.caymanchem.com/pdfs/10006315.pdf
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I am looking for help on how to formulate a phrase.
I have excluded patients with entirely missing data, but have included cases with only incomplete documentation. How do I include this in my paper and express that therefore statistics and reported results do not align with the total number of included cases?
And where would you advise I put this information? Under the Methods section? Discuss it under limitations?
Thank you, I appreciate your help!
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Best methods suitable for isolation and purification of plant pathogenic fungi and bacteria from plant and soil samples....Latest methods....Any special advantages....Procedure to be followed...etc.
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For isolation, fungal media with antibiotics should be used to suppress bacterial contamination. Sabouraud's dextrose agar and potato-dextrose agar are commonly used media. Cultures are routinely incubated at 25° to 30° C for up to 4 weeks. Isolation of zygomycetous fungi can be difficult... https://www.sciencedirect.com/topics/immunology-and-microbiology/fungus-isolation
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I am organizing some materials for the MSc and PhD students and want to categorize the references in some dimensions so the students can choose by their own.
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Quantitative Applications in the Social Sciences. A sage University paper series. This is an excellent collection of 175 brief volumes addressing quantitative topics such as Regression, Models, Data Analysis, Structural Equation Modeling, Experimental Design, Factor Analysis, Measurement, ANOVA, Survey Data, and many more areas of interest to faculty and students alike.. I had the entire series and lent them to students on a borrow one, return one arrangement.
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Please enlighten and attach the reference of past studies.
thanks
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Cohen's book, Statistical Power Analysis for the Behavioral Sciences, was a ground breaking text on the determination of sample size. G*Power is simply a statistical program that implements and extends Cohen's original ideas.
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I am specifically interested in works of art for which ratings on multiple dimensions are readily available--for instance, ratings of beauty or of liking that were acquired from a sample of volunteers. Features of interest also include the actual features of the artworks themselves, like brightness, complexity, etc. I have been pointed towards several options:
  1. Prediction of beauty and liking ratings for abstract and representational paintings using subjective and objective measures (https://osf.io/2sy4f/)
  2. JenAesthetics (https://www.inf-cv.uni-jena.de/jenaesthetics.html)
  3. The Strohminger Grotesque Art Database (https://ninastrohminger.com/grotesque)
I wondered whether there might be others I am missing? Thanks, in advance, for any help anyone is able to provide!
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Not specifically art work, but this is a fairly large database of beauty ratings on real outdoor views of the UK.
Dryad Data -- Using deep learning to quantify the beauty of outdoor places (datadryad.org) https://datadryad.org/stash/dataset/doi:10.5061/dryad.rq4s3
ScenicOrNot (datasciencelab.co.uk)
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I plan on doing cell culture on Thermanox cover slips so that I can take TEM images of my monolayer cultures. I've seen that I can place the cover slips into well plates for culturing purposes, but I don't know how to go from "I have my cells in suspension, ready to be seeded" to "the cells are now seeded on the cover slip".
Do I have to be very precise with where I pipet the cell suspension? Do I just fill up the entire well with the cell suspension? Any insight would be very helpful!
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You could follow the same protocol as you do for the normal seeding of your cells.
(I always autoclaved the coverslips prior to seeding cells on them to avoid any contamination.)
You can use forceps to place the pre-autoclaved coverslips on the cell culture plate (also to take them out once you want to process the sample for TEM).
If your cells have problems to attach onto the coverslip then you can also pre-coat (45min at 37C) the coverslips with Poly-L-Lysine or Matrigel (for better attachment) prior to seeding the cells.
For the pipetting, I had optimized the cell number to a specific density of cells so that they would easily attach in every part of the wells and coverslip as monolayer.
So I did not have to pipette precisely.
Hope this helps!
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As all of us are familiar with the different journal ranking systems and requirement conditions, in many cases we meet different kind of fees, charges for publishing our researches. Usually only the submitting and the previewing cost US$ 50-250, which is non-refundable and the paper may be rejected by the editors without being sent for review. Others introduce fees for the publishing US$ 500 -1000 (extra fees for colour graphs, maps etc. or for appeals against a Chief Editor's decision). For good English, they offer some links for the grammar review: US$ 100-200. Besides all of this, they employ embargo for 12-36 months, and ask US$ 600-2500 for open access. I think these fees sometimes unreasonable, so it is hard not to find the business factor behind them.
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I do agree with the expert answers above
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Hi All,
As it says in the title really. From what I can find online, a lot of literature focus on just OM proteins/vesicles rather than whole out membrane preps. So if anyone has some papers they wouldn't mind sharing, or know if there's a specific search phrase I should be using, I'd really appreciate a comment :D
Many Thanks in advance,
Peter
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Hello
One excellent reference for different membrane isolation is Methods in Enzymology Volumes 20 & 22. I recommend that you have a look at it and I think you will find in it what you ask for.
Good luck
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I am conducting a Large N fsQCA, and I need a basic bibliography on this topic. Especially on scope conditions, necessary tests, and its relation to the deterministic ontology of the method. Would someone indicates it for me, please? Thanks!
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The librarian of your institution may help you find out which specialised literature databases / electronic journals are available to you. In addition, you can use Google Scholar to search for scientific articles: https://scholar.google.com. For example, this is a recent article (2020) I found as one of the first hits searching for "Large N fsQCA" (including the quotes): https://maggetti.org/wp-content/uploads/2020/06/Thomann-Maggetti-Designing-Research.pdf
While I know nothing of your particular field, the journal title (Journal of Social Research Methodology 19:445-459) seems promising if you look for information on the methodology - you may check on the impact factor of this journal to find out if it is a reputable journal in your field. Combine different search terms to focus on the subfield you are most interested in, follow the references cited in articles that seem relevant, search for more articles by the same main authors.
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Hi everyone,
When looking into thematic analysis, you always find a notion of it as a method rather than methodology. Therefore, I was wondering, if TA can ever be considered as methodology? If so, under what conditions, and what philosophical stances does it represent?
If it's always just a method, is there any particular methodology that links with it?
I hope this makes sense. Thank you for your replies.
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Marketa - thematic analysis is a qualitative method. In terms of methodology - it can be a method adopted by many qualitative methodologies - although there are exceptions. For instance, grounded theory uses its own analytical discourse - and various phenomenological approaches use and adopt their own specific analytical frameworks/methods. As David and Marek have correctly identified - TA cannot be a methodology in its own right. It is a 'tool' - rather than a philosophical/theoretical 'lens'.
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I have been trying to estimate some biochemical parameters in urinary bladder but homogenization is problem with me as indicated by very low amount of protein in homogenate.
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Dear colleagues,
I am working on an intervention study with 3 different groups of students. One group represents the intervention group and consists of a seminar with a practical phase and computer science content. Another group has only a theory seminar with computer science content and the last group has a seminar with other content. The last group is used to check how stable the constructs are. The 3 measurement points are equally spaced as pre-inter-post-tests in a quasi-experimental setting. Latent growth curve modelling doesn´t fit! Is there a method that uses the strength of the 3 measurement time points with a small sample size?
Thank you
Martin
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I would use a repeated measures ANOVA for Between x Within subjects designs.
In attachment a script using R for such a design, with Post Hoc tests. In the example there are 5 observations in each Condition.
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I am going to use this method to load protein hydrolysates into milk phospholipid nanoliposomes. However, I couldn't find a lot of information about this method. I'd appreciate it if someone can provide me some more information on this specific topic.
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Check this Patent which explains method and apparatus for manufacturing drug delivery vesicles (including Liposomes, Nanoliposomes, Tocosomes, Niosomes, Solid Lipid Particles, Lipidic Nanoparticles, etc.) in a safe and environment-friendly approach:
Mozafari, M. R. (2005). Method and apparatus for producing carrier complexes. UK Patent No. GB 0404993.8, Int. Appl. No. PCT/GB05/000825 (03/03/2005), 14.
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A few days ago a colleague of mine made me think about the impact the COVID19 crisis will have on cohort studies. Especially those focused on causes of mortality and the elderly will be deeply impacted by the number of deaths due to the pandemic.
Is this something manageable?
How big is this matter in your mind?
How can this be handled?
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Well, the cohort study is highly suggested in covid pendamic. however, the covid-19 is no more a cohort situation it is done for observation and per and post-test by training and other medication.
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I need di- and tri-peptides for my intended work. I was going through internet and came across three different methods..1) by Solid state peptide synthesis (SSPS) 2) Liquid Phase Peptide Synthesis and 3) by Bacterial expression....However for first two..we dont have lab facility...so I wanted to know whether Bacterial expression could be carried out for synthesizing small peptides? Which one would be better?
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Sir
No doubt for di-tri-tetra up to 15amino acids sequence means only SPPS method was best .
Bacterial expression method only for long peptides and naturally available Protein expression purpose - also for this you need to do lots of ground work and also need perfect analytical setup labs. not able to find target di-tri-tertra peptides by this method, more complex.
Liquid Phase method was ok but for small quantity research purpose SPPS was better than LPPS,
Cost wise low, so you may get it from Custom Peptide Synthesis Labs.
We too providing the same service.
All The Best For Your Research & Success
- Dr.R.Selvam - Custom Peptide Synthesis Expert
Grey Matter Research Foundation.
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I have a data which consists of an excess of zero counts. The independent variables are number of tree, diameter at breast height and basal area, and the dependent variable (predictors) is number of recruits (with many zero counts).
So, I want to use Zero-inflated negative binomial model and Hurdle negative binomial model to analyze. My problem is I do not know the code of these models in R package.
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Dear Kaushik Bhattacharjee
thank you for the useful link, thanks a lot
best
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I am in need of a free on-line software for performing chemometric analysis and chromatographic alignment of retention time shifts. any suggestions!!
Also i have another inquiry about how the numerical data matrix of HPLC-MS/MS can be arranged? I mean how the data of time (min)/relative intensity (%) for MS, TIC monitored chromatograms can be composed in an excel spreadsheet file to be imported into the software?
Thanks in advance
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Hi, I am affraid it is way too late to answer this but for the record...
You should consider learning the fabulous statistical computing and graphical representation R. R is freely available for several operative systems and contains several libraries specifically designed to solve many data analysis questions. Here is a shortlist of some libraries you could use:
Chromatographic data treatment
Chemometrics
A good introduction to R may be found at https://stat545.com, https://www.statmethods.net/, and many resources more.
If you dive into a non-programing-like software you may need to spend some time learning how to use it anyway... If you spend that time learning R you can use it for many other purposes that will enrich the way you treat and visualize your research data so it is worth giving it a try.
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Do kits such as the NEBNext microbiome DNA enrichment kits really work for the enrichment of (all) viral DNA and how species specific is the host DNA removal (I'm working with bovine blood samples)? Any practical experience?
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Hi, did any one try with Genomiphi™ V2 DNA Amplification Kit ?
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Hi everyone,
I'm trying to find a home for an instructional manuscript on how to use NVivo to perform qualitative methods (abstract below). I've tried three different journals (Current Psychology, Psychological Methods, Qualitative Health Research) with no luck (they say its good content, but not a good fit). Any suggestions?
-David
Abstract:
From 1995-2016, there has been a 15-fold increase in qualitative scholarship in the social sciences but the rigor and quality of published work has ranged widely. Little scholarship provides concrete, pragmatic explanation of (and direction regarding) execution of systematic, high-rigor qualitative analysis. The present article guides the developing qualitative researcher through technical and procedural aspects of analyzing qualitative data with specific attention to reliability and rigor. Guidance addressing transcription, importing data, forming coding pairs, performing initial/open coding (examples of three types), determining core themes, systematic team-based coding, maintaining a data audit trail, creating a Numeric Content Analysis (NCA) table, and preparing work for publication is provided. Material includes several tables and figures that offer practical demonstrations of how to use Nvivo in data analysis. Transcription tips and outsourcing benefits and cautions are also offered. Altogether, the present article provides qualitative researchers practical guidance for executing multiple stages of qualitative analysis.
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Not everything is suited for publication in a journal. Perhaps someone is working on a book on qualitative analysis, and would be interested in publishing it as a chapter, or... maybe it's time to do your own, and find others to write some chapters.
Another option would be to self-publish on the web. There are a number of options for that, or you can simply create a website for it.
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I want to quantify the amount of D2O and HDO present in water below 1% level. 
1) For now, I only need to know the amount of deuterium present in water.
2) But, it would be better if I can analyze the ratio of D2O, HDO and H2O .
Can you recommend an analysis method? Simple method is better.
Thanks
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Hi, we use FTIR
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Hello!
I'm gonna start to work with HepG2 cells and I would to know what are best culture conditions for these cells. I have been reviewed some articles and people use many types of medium and additives.
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ATCC
DMEM
Not important all can use.
Combination media of DMEM and RPMI 1640.
What cells do you want?
What drugs do you want to test?
All you can do it.
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I worked on antibiofilm activity of some enzymes by crystal violet assay on 96- wells , but what's the best way to destaining CV 95% ethanol or 33% glacial acetic acid and why ?
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What is the procedure of biofilm and antibiofilm assay
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If i want to study the experiences of ( students, teachers and administrators ) related to a new course by using a questionnaire with students and teachers and interviewing the volunteer teachers and the administrators.
So, I will use a quantitative design with students
a Quantitative and qualitative design with teachers ( I will interview just who are write his/her contact information on the survey tool) and
a Qualitative design with administrators.
Can I describe this method as a concurrent design?
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I support previous comments.
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Hi:
we are working on isolating bacterial strains that produce secreted cholesterol oxidase using a minimum medium with cholesterol as the sole carbon source. I noticed that the cholesterol forms an emulsion and obviously doesn't like to dissolve well. So the bugs might see very little cholesterol in solution. This problem is not mentioned in the paper we follow.
The questions are: a) does that matter? and if so b) What do I do to get cholesterol dissolved? What would be a good detergent to use without killing the bugs?
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I know this is very old but I love it because it screwed many conclusions: using tyloxapol with ethanol in chemically defined media with sole carbon sources is a terrible idea. Ethanol IS a carbon source! There are plenty of NAD and quinone-dependent ethanol dehydrogenases able to convert ethanol to aldehyde. From there, is a small walk in the park to central carbon metabolism.
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" Pitfall trapping is the standard method for collecting ground-dwelling arthropods and soil fauna in studies of ecological and agricultural entomology " ( Ruiz-Lupión et al. 2019).
In my current research assistant position I am working on analysis of macro-fauna in forests. We use pitfall traps to assess the abundance of macro-fauna in a given area. I'm curious to learn more about other methods used for this sort of analysis.
  • What methods for pitfall trapping have you used, if any?
  • What were the advantages/disadvantages and what would you have changed about the method you used.
Our methods are as follows:
  1. Briefly, we plant a plastic cup in the ground with a cover on top (to make sure mammals or larger animals do not enter the trap but only macrofauna can enter)
  2. we leave the cup for several weeks
  3. The macrofauna fall into the cup and are preserved by antifreeze, which are then taken into lab for identification and abundance counts
  4. By measuring the area of the cup's top, and how many bugs have fell into said area, we can then gain a better understanding of the abundance of macrofauna in the area
In a study reviewing pitfall traps, Ruiz-Lupión et al. (2019) states the factors which should be considered by ecologists using pitfall traps. They state, "the capture rate of arthropods in pitfall traps is proportional to their activity, and the number of individuals that each trap catches may or may not reflect their true abundance, and instead just their activity. Thus, the rate of capture is proportional to the joint effects of abundance and activity, something that has very often been overlooked by ecologists for a long time... [Nonetheless,] activity estimates from pitfall trap catches can still be biased because of multiple factors such as the surrounding habitat structure or the environmental conditions such as temperature and water availability. Additional factors could be the vertical distribution of the soil and leaf litter layers, as well as the attraction or repulsion of preservative fluids, detergents, or baits, the effects of which vary according to the taxon, sex, season, and environment. Specifically, if a trap retains excessive amounts of water, it could act as an attractor for the fauna, especially during drought periods, therefore biasing the estimates of activity. "
References:
Dolores Ruiz-Lupión (2019). New Litter Trap Devices Outperform Pitfall Traps for Studying Arthropod Activity. Insects 2019, 10(5), 147; https://doi.org/10.3390/insects10050147
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This is on RG now. Fitzgerald L.A. 2012. Finding And Capturing Reptiles. Pp.77-88. In R.W. McDiarmid, M. S. Foster, C. Guyer, J. W. Gibbons, and N. Chernoff (eds.), Measuring and Monitoring Biological Diversity: Standard Methods for Reptiles. University of California Press, Berkeley, California.
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I want to calculate Potential evapo-transpiration with less data requirement so which method can be used for this which will give accurate and appropriate results.
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Hello Dr Shruti Bansode
The evapotranspiration is a fundamental variable in the hydrological cycle and is a key factor for water balance and irrigation.
The penman monteith method is more accurate than the Thornthwaite method because it takes into account several climatic parameters including air temperature, relative humidity, wind velocity, sunshine duration, etc.
See the attached file for more details on the penman monteith method
Best regards
Fatah BOUTELDJAOUI
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Hello!
Hope everyone is doing alright during these turbulent and uncertain times.
I am working on my masters dissertation (MSc International Management) and would like to answer the research question of "how adaptable are frugal health care systems in developed markets?" (such as U.S.). I am really stuck as I cannot get hold of any experts to interview and data/literature is limited, as I have found so far. Does anyone have an idea of how I could tackle the methodology or has any tips on where to get data from in this area.
P.S. : Would not mind changing the angle on research question in order to make finding data/literature easier.
Many thanks in advance, feel free to contact me, if you have further questions.
- Juliana
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Dear Juliana,
Unfortunately, my answer is rather suggestive than a problem-solver one as I am not an expert in the mentioned field. However, seeing empty discussion motivated me to share some basic ideas that came into my mind. If you have such a narrow research interest, expert interviews are really important to implement a method to study certain phenomena. My recommendations are as follows:
  1. Try to use alternative or synonyms to expand and to enhance the literature review. Presumably, frugal health care systems depend on certain pre-conditions like innovations and government policies. So, you can adjust your research question or literature review directions to more expanded horizons because otherwise simply you can not start and finish the research;
  2. In my opinion, you can benefit from the agents who provide frugal health care. Instead of an interview, you can also conduct an evaluation via surveys on the limited sample size. I am sure there are available methodologies that allow an analysis of the small sample quantitative and qualitative data.
  3. Moreover, there are actors and players behind the scenes related to frugal health care systems in the developed markets. Maybe, you should look for them.
  4. Maybe there are some mediators, they are not directly involved but they can help to track some things.
  5. Always collect and survey the professors' and colleagues' opinions and ideas.
  6. Find dissertations or theses that at least indirectly addressed the topic you are interested to get a general framework of your study.
Best Regards,
Ibrahim
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How will 4% w/v SSA interfere with the NADP/NADPH ratio? Can this method of deproteinization be used as an alternative to 10 kDa cut-off spin columns in order to measure NADP/NADPH ratio?
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Good Answer Achim Recktenwald
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Hi everybody. I am facing an issue about reproduction data analysis in chronic toxicity test with C.dubia. US EPA Method 1002.0 ( https://www.epa.gov/sites/production/files/2015-08/documents/short-term-chronic-freshwater-wet-manual_2002.pdf) instructions tell:
" The response used in the statistical analysis is the number of young produced per adult female, which is determined by taking the total number of young produced until either the time of death of the adult or the end of the experiment, whichever comes first. An animal that dies before producing young, if it has not been identified as a male, would be included in the analysis with zero entered as the number of young produced "
Including dead animals in the count gives me back too high standard deviation values which don't fit well in my analysis. I would like to know if alternatives are available: feel free to link me to other papers, works, methods etc.. which could help. Thanks a lot to anyone will spend time to help me.
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Hi!
I graduated with a Neuroscience and Psychology BSc in 2016, and my final year project was a bench-lab in vitro study of omega-3 oils applied to fluorescently-stained cortical neurons. I realised that although I had a passion for neuroscience, my first foray into applied research was not promising; I hated the tedium of the process.
I then did an MSc in Mental Health Sciences, with my thesis originally using an EEG/MEG dataset on psychosis patients. Again, I loathed trying to learn (the tedium of) Matlab and learnt instead that I'm fundamentally not a coder.
I changed to studying the mid-term after-effects of a psychedelic drug using a battery of questionnaires. I loved this psychometric approach, and it was easy enough running simple statistical analyses. However this was firmly psychological, not neuroscience.
Now I'm doing a PhD largely employing thematic and other qualitative analyses of altered states, which I thoroughly enjoy. But again, I'm still attempting to retain some know-how in neuroscientific methods. I'm doing some basic secondary EEG analyses on the aperiodic signal - though not loving even the rudimentary coding necessary.
My question is what are some other neuroscience techniques available out there which I may be more suited to? Can one even be a 'neuroscientist' without having to use a wet-lab - or matlab?
Perhaps tDC or tMS? Could someone maybe elaborate on the types of questions that can be answered with these; or the analyses that are run after them?
Other than f/MRI, which I assume always require programming - do other neuroimaging modes e.g. PET/SPECT also require this?
I'd also be very interested to hear people's thoughts on/experience with working to incorporate as much understanding of neuroscientific findings in strictly psychology studies - While writing psychology papers, referring strongly to the neuro literature to inform and discuss the study's aims and findings?
Thank you very much,
I'd be really appreciative of any insight :)
Pascal
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I am doing a plaque assay with HeLa cells and infecting them with poliovirus. I dont get plaques?
Method i use:
1. I take a 80-90% confluent plate of HeLa cells. I do the assay in the 6 well plates from Nunc.
2. Dilute the virus in serum free DMEM-10-2 through 10-11.
3. Wash the plates with 1X serum free DMEM.
4. I infect the cells with virus 300 ul for 30-50 minutes at RT/rocking.
5. Overlay with 5ml(2XRPMI without phenol red) + 2.5 ml 1.5% Agarose +2.5 ml 1.5% Agar.
6. keep in the 37 degrees incubator, 5% CO2 for 48 hours
7. Fix with 10%(v/v)formaldehyde in 1XPBS for 30 minutes.
8. Stain-0.5%(w/v) crystal violet in 10% EtOH for 5 minutes. and wash with water.
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For Poliovirus use BGM cells ( Buffalo Green Monkey cells)
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We want to do a study that test if the placement of your mobile phone effects your memory. If it being next to you, decreases your memory by disturbing your attention. So the phone will either be next to you on the table, in your pocket/bag or in another room.
Is it necessary to have multiple complex tasks, such as the operation span test and another one or is that one enough?
As we are quite short on time, studying long term memory is not possible.
Thank you for your answers.
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There are studies on the subject on the Internet, you can benefit from.
my best wishes.
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I am a first-year PhD student who would like to hear about other researchers' experience related to interdisciplinarity within the humanities, especially those combining both text and visual elements:
  • Which are some suitable methods that can be applied?
  • Should both areas of studies be represented in a balanced way?
  • Any tip that you would have loved to hear before developing your own interdisciplinary project
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This largely depends on the more specific areas that you are researching of course. There are correlations between literary theory and art/aesthetic theory, and both can and are often utilized for art and literature, so you can draw correlations between the two. In addition, I tend to use theory that already exists in the interdisciplinary realm, like Derrida’s hauntology, or Kristeva’s abjection(two of my favorite). The balance between the two fields Is really up to you and how you approach your research as long as one isn’t an afterthought, in which case you need to question whether it belongs in the project. I hope that helps!
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I am interested in relationships between photovoice or auto-photography as research methods and social-spatial difference, either as captured in the photographs, or as embodied or lived by the participants. I would particularly appreciate suggestions of literature from the past 10 years.
Recommendations of reading on participant-photography and social-spatial difference would also be relevant in this case.
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Thank you!
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I am currently developing a project where the primary mode of inquiry is rooted in practice based methods and techniques. The project is focused on a social dance form. I have read several books/articles in which practiced based methodologies are used, however, I am wondering if there are methods/techniques out there that they better satisfy my project. What practice-based methods/techniques may be beneficial for examining social dance styles?
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Hola. Si lo que deseas es conocer las perspectivas que tienen las personas acerca del baile puedes trabajar con el método fenomenológico o con interaccionismo simbólico. Las técnicas básicas son la entrevista no estructurada y la observación directa o la observación participante
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is flconazole really soluble in water...? in which tybe of water? should it warm or normal?can we dissolve it any other solvent like DMSO in place of water,to determine it's MIC.
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Poorly soluble in water but can be dissolved in organic solvents such as chloroform, propylene glycol, and polyethoxylated castor oil
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There are several methods for calculating Mixing Height but requires upper air data. Is there any source for finding mixing height using surface met. data. Kindly help me to find them.
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What is the best method for preparation of 10 % tissue Homogenate?
I used to homogenize the tissue in Ice-Cold Saline.
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weight the tissue sample and add 9 times the volume of pbs according to the ratio of weight: vol
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Distinguished colleagues,
I am going to update my methods of delivering lectures at university. Any publication link or comments on this regard will be highly appreciated!
Thank you in advance!
Best regards,
Dr. Vardan Atoyan
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I agreed with Jose, beside depends of the courses also the participant (initial or intermediate or advance), in my opinion, Integrated and interactive approach in learning process, collaborate of Active, creative, effective and enjoyable learning. You may try kind of teaching method (materials) some of ideas refer to your objective or purpose, example: story telling or gaming or quiz. Well good luck on your project..Cheers
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Distinguished colleagues,
I need your professional opinion for my ongoing research. Any input, support, publication links or comments will be highly appreciated!
Thank you in advance!
Best regards,
Dr. Vardan Atoyan
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My favorite ones are comparison, case studies, and interviews.
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What are the methods, techniques, lines of thought, university labs considered vanguards in hydrogeology?
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Interesting that you should ask this. I have been pondering the subject myself. My conclusion is that our computer abilities exceed our physical hydrogeology skills. We have wonderful models that have ridiculous parameters for real-world materials, yet by manipulating these parameters some semblance of the real picture emerges. What if we actually built models with real parameters and manipulated the model code to produce a model?
Secondly, hydroGEOlogy implicitly involves the earth. HydroLUNology, HydoARESology, etc. - these are the subjects that new hydro___ologists will explore. What fun!!!!
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Hi -
Has anyone ever had the issue when they are doing Loss on ignition analysis in a muffler oven at 1000C? Previously I worked with a newer muffler oven and had no issues weighing and baking soils at 1000C. I recently started using an different muffler oven which is a little older, but is properly functioning. I baked my soils and crucibles at 550C and it was fine. I then turned it up to 1000C and baked it at 1000C for 6 hours and turned it down to 105C to take the crucibles out after cooling. However, after I took the crucibles out all of them seemed to be changing colors from neutral ceramic white to green from the bottom up. Also it looks like there are small crystals or precipitate forming on the crucibles, more densely from the bottom up. Attached are a couple of pictures. Does anyone have any experience with this or have any suggestions on how to get to the bottom of this or fix it?
Edit: I just want to note I tried to clean these crucibles with an HCl acid bath, and the greenness did not go away and the precipitate crystals seems to arise once it is fully dry again.
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I have also observed this. At elevated temperatures, some elements will dissolve into the solid solution of the crucible. Look into the ceramics literature on vitrification. I would not worry about the color change, nor would I be concerned about contamination unless you are ashing a sample for subsequent digestion and quantification of elements in the ash. If you're simply looking at soil carbon, the altered crucibles will continue to be fine. Disregard the etching and color change. Keep in mind though that some elements like potassium volatilize at high temperatures (> 500 C).
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I would appreciate any suggestions for effective methods for teaching geology to a blind student. 
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Geology can be heaven for blind people.
1) Let you think what is possible to do with touching minerals with different shape (large mica flakes, pyrite cubes, amphibole prisms, magnetite octahedra, leucite icositetrahedra, garnet dodecahedra, calcite scalenoedra, and so on).
2) Fossils are even better. In particular, it useful to let blind people touch objects with the same shape but with completely different origin. For example, let you think to a cone with variable heights and radius/height ratios. It could be (and you can show and give in hands): 1) belemnite; 2) hyppurites; 3) horn; 4) tooth; 5) coral; 6) hybodus and so on. All these fossils can be found in e-bay for a few dollars/euros and you can talk for half an hour in describing all the fossils and their use.
3) Moreover, you can let blind people play in handling teeth of different origin (ursus speleus, ancient horses, spinosaurus, plesiosaurus, basilosaurus, carcharodontosaurus, mosasaurus, otodus, megalodon, mammoth, onchopristis, swordfish, wholly rhynoceros, enchodus, gomphoterium and so on). With less than 500/100 US$ you can buy one or more pieces of all these teeth.
4) You can 3D print scaled planets and Sun, to show them the relative size of the planets.
5) You can show also 3D plastic charts showing bathymetry and topography.
6) Rocks and minerals with different softness and hardness (e.g., graphite or graphite schists, talc or talc schists, granites, obsidians, greywacke, siltstone, and so on).
7) A normal Estwing hammer.
8) Iron and silicate meteorites, to show the different density of the rocks, with a pleasure to touch extra-terrestrial stuff.
I have served as director of the Earth Sciences Museum of the University of Rome (MUST = Museo Universitario di Scienze della Terra) for three years, and we have used these styles of interaction for blind people, receiving excellent feedback. Now I came back to my usual business that is igneous petrologist.
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What is the suitable method for disposal
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Every country has its waste classification criteria. I some countries it is classified as non hazardous waste and in europe , US and in some other countries it is classified as hazardous waste. If EAF dust has more the 16% zinc then you can easily get rid of this dust as there are many buyers are Available in the market for zinc recovery and they can easily pay you between 50 to 100 usd / ton .
In case if zinc content is less than 2% then it can be used in cement plants and ready mix industries. In case if you need more information then please contact me on my email if [email protected]
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Can anyone please help me to find all the biochemical tests for bacteria?
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Find the biochemical test of 60 bacteria in a single page.
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Our lab looks at tumor growth in the bone microenvironment, and in doing so, I section mouse tibia on a fairly regular basis. These tibia have been manually cleaned, fixed in z-fix for a total of 48h (24h at RT, change z-fix, 24h at 4 degrees), then decalcified (10% EDTA, pH 6.5) for a total of 2 weeks at 4 degrees (changed after 1 week). They were then rinsed well with deionized water, put into 70% ethanol, and sent to Histoserv for paraffin embedding and initial sectioning for H&E sections. I then section these blocks for addition staining, and most of the time, everything is okay. However, I come across some blocks where, for some reason, part of the bone will chip out during the sectioning (as the blade goes through the block, you can actually see the chip of bone come out). I have tried to re-paraffin these blocks by softening them, placing the chip back in place, putting a small layer of paraffin on top using a spatula, then re-molding the block. Sometimes this works, sometimes this doesn't. Does anyone have any suggestions for what to do when the bone chips out of the block? Thank you!
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Hi Mackenzie,
You can try following steps:
1. Use higher EDTA concentration (15% at 4℃ for 2 weeks with change every alternate day) to make the tissue soft enough;
2. Trim unnecessary part of the bone to make it smaller size before decalcification;
3. Use sharp blade for sectioning.
Hope it will help to fix your problem.
All the best.......
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I plan to do an anthelminthic assay using Shorea leaves for my BS thesis and I consulted vets and nurses (unfortunately, we don't have a parasitology department or even a dedicated parasitology professor in our school) and have obtained a wide and contradicting opinions on the matter. I want to get results as close as possible to reality and so I plan to use A. suum as opposed to using C. elegans or P. posthuma. A vet I asked suggested I do an in vivo assay using lab rats as host and T. muris as model organism or A. galli on chicken host. Additionally, they said performing in vitro assay will invalidate my results as the conditions in an in vitro set up may be too artificial and may not reflect what is happening inside the body. One of the nurses I asked suggested I skip using model organisms altogether and use in vitro assay using A. lumbricoides or an in vivo assay of the same organism but on a guinea pig host.
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Anthelmintic drug development includes pre-clinical research ( in vitro studies) and clinical studies (in vivo studies). Scientific validation of the anthelmintic properties of plant extracts is evaluated mainly through in vitro assays, followed by in vivo studies. Considerable physiological differences are expected to be present between the in vitro conditions and in the predilection site of the endo parasites within their host animals, including factors within hosts that affect the bioavailability of the active compounds. In vivo studies are more relevant and more reliable. The results obtained in vitro should be substantiated by in vivo studies.
Don't use P.posthuma as animal model for in vitro studies. It is an annelid living in the terrestrial habitat, the physiology of which differs from the endo parasites. Use only helminth parasites
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I want to use a surface again, that has already been mounted with prolong antifade mountant and am looking for a removal method for this. 
Any help is much appreciated
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I just tried this method and put the mounted glass slide in 37°C PBS over night. Next day the cover glass was detached and the mountant completely dissolved. Late answer but a confirmation for others.
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Hello,
I have two methods I want to compare, however I have different sample sizes in each method. One has 12 samples and the other 25. The idea is to see if collecting more samples gives better results.
I was thinking a Bland Altman but don't you need the same sample sizes? Do I just do a normal unpaired t-test for this?
Kind regards.
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I disagree with Peter's advice about testing for normality.
The normality assumption for the unpaired t-test is that the populations from which the scores are sampled are normal. Unless the outcome variable is something that has never been studied before, there is likely lots of information beyond that which is available in the two samples in hand that can be used to judge whether the populations are approximately normal. This is one of the main points in the following BMJ Stats Note by Bland & Altman:
Another major problem with testing for normality prior to conducting a t-test is that tests of normality lack power when sample sizes are small, when the normality assumption is most crucial; and they have too much power as n increases--and as n increases, normality becomes less important (because the sampling distribution of the difference between the two sample means approaches the normal distribution). Here is a short conference presentation I gave on this topic a few years ago.
K. Hopton, I think it would help if you gave much more detail about what you are trying to do. Bland-Altman agreement analysis requires paired measurements, but you appear to have two independent groups. What are you measuring, how are you measuring it, and what do you hope to find? HTH.
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It should be estimating total mineral content as iron. Does anyone have suggestions?
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yes i need sir. i am doing the research
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I want to detect 50-bp segment of the recA gene Bordetella avium. Seqence of probe and primers: 5-CATCGCGCTGGGTG-3 NED fluorescent reporter on the 5' end and nonfluorescent black hole quencher at the 3' end. Primer forward CGGTTCGCTGGGCTTGG and rewers CACGCGGCAGCCCGC. Can I change probe dyes to FAM on the 5' end and on BHQ-1 on the 3' end without losing specificity of reaction? I will be thankful for any help
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BhQ2 is best, you should be guided by the emission peak (for dye) versus absorption peak for (quenchers)!
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I am having issues transferring my large protein (220kDa) at 30V for 2 hours (RT). It is often patchy with incomplete bands (annoyingly I occasionally I get a clear transfer!). For my other protein at around 40kDa (for b actin) I have no issues.
I have read that for larger proteins people either run overnight at 4 degrees at 20V or for 3 hours at 70V.
Ideally I would like to run in the shortest time possible! One of the reasons why is the power pack i use is pretty old and I don't know how well it would withstand the cold room (rusting)!
But my main worry is losing the smaller proteins if I up the voltage for a prolonged time.
I am using pre-cast tris-glycine 4-20% and the transfer buffer is composed of 10% methanol, and tris-glycine buffer (x2) for a wet transfer.
I was wondering what protocols work for you or if you have any suggestions on what I should try first?
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Dear Olivia,
I have transfered proteins up to 250kDa and tubulin (around 50kDa) at 100V for 1 hour.
I put the transfer system in ice and I use a precooled transfer buffer ( 20% methanol, and tris-glycine buffer (x1) ).
I hope you find it useful.
Erica
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Hi all,
I am currently comparing two groups (control/exp) using SEM. Each latent construct (5 in total) has at least 8 if not 20 individual indicators, as such I am using appropriate parceling techniques in each. The first step requires I conduct a CFA for each construct of interest - to do so, I must have at least four indicators. In the full SEM, as there are 5 latent constructs in total, to improve overall fit, I wanted to reduce the number of parameters to be estimated to be two indicators for reach latent, rather than three. My question is, is it permissible to run a CFA with three indicators for each individual latent, but in the final SEM - only two indicators (using an appropriate parceling technique - rather than just dropping items).
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Hello William,
I can see where you are going on this. Usually a CFA for a SEM measurement model includes all factors with their indicators. There is no structural model and the factors are simply correlated to each other. This overcomes the issue of “needing” at least four indicators. Yes, you can run a CFA with only two items. You can even use a single item with modelled residual variance if the reliability (Cronbach alpha) is well accepted in literature. For myself, I find that factors with two items often throw up Heywood cases (problematic). I tend to use 3 items to ensure the factors are just identified, and usually require less constraints (as Matthew noted previously).
Once you have a “good” fitting CFA, you can move on to the structural part of the modelling.
To your second question, it is not advisable to confirm your measurement model (CFA) and then use different measurement items; you are not using the measurement model you confirmed. Furthermore, your next step beyond the measurement model for the complete dataset is to test measurement invariance between the control and experimental groups you are comparing. This is a critical step for your SEM analysis and involves ensuring that the latent constructs are understood by both groups in the same way (i.e., they mean the same thing b/w groups).
Chapter 9 & 16 in Kline’s book are good intros to CFA and measurement invariance, respectively. See: Kline, R. B. (2016). Principles and practice of structural equation modeling (Fourth ed.). New York: The Guilford Press.
Wishing you well in your research,
Tim
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I am looking for any insight or proven practice regarding the usage of email for research. Is there a program/website/process to anonymize both a researcher and participants' email addresses but still allow them to communicate?
Craigslist has a feature where the buyer and seller can email each other using auto-generated, anonymous email addresses ([email protected]). These emails go to the buyer/seller's main email accounts, but they strip the messages of any headers/identifying information. In effect, these emails through Craigslist are anonymous.
Is this something we could utilize for research? Could we have participants email a generic email address that would allow both the researcher and participant's real email address to be redacted, yet allow for communication?
Has anyone else tried something similar?
Thank you!
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I had a questionnaire with 18 close-ended items about factors of students' failure in English courses.
At the end of the questionnaire I asked respondents an open-ended question:
Why did you fail English in the previous semester(s)?
I made it optional and only 27 out of 56 responded to this question.
Was it correct?
Regards!
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It's certainly OK to ask open-ended questions after a set of closed-response questions, but in your specific case your question might have been a bit too abrupt to encourage responses. Many of your respondents might not have known the answer, or the answer might have been complex. If you had asked in a slightly different way, perhaps you would have received more answers. Furthermore, there might have been different reasons for failure in different semesters, and participants might have considered it too burdensome to go into details.
I wonder whether, with a bit of piloting and/or careful thought you could have asked for the information that was of importance to you in another closed-response question.
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I want to use the contingent method, is there any researcher have used it and can give me an idea regarding the methodology and required data for this method.
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Respondent analysis for every services and then Delphi
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Hello everybody! I have 3 research problems and I will be using Colaizzi's method of data analysis. I am fairly familiar with the steps of Colaizzi's method but I do not know how to start it. Do I analyse my transcripts as a whole to get a final thematic map and be able to answer the 3 research problems or do I have to sort out the significant statements relevant to each research problem and then later arrive to 3 different thematic maps?
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Hello Apri,
I did not know the answer, so looked for help on ResearchGate. In this article:
Prof Suryani Suryani describes how she started. After the translation was carried out, on the fifth page, in paragraph 'Extracting significant statements' she describes how she dealt with the transcripts. You could always send her a message if you need help.
I have not seen the full text, but it might help. Prof Suryani writes, when referencing Sanders, 'Colaizzi’s method of phenomenological inquiry comprises seven steps' (3rd page).
I thought this might help:
Full text:
but I'm not sure.
I hope this has been of some help. I hope someone responds who has used this method.
Very best wishes,
Mary
.
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I'm trying to make these similar concepts clear in my mind but it's not easy. Let's make a brainstorming together!
Although there are some differences the related literature says:
the intent of a meta-analysis is to aggregate the findings of quantitative studies and analyze this for a true effect;
the intent of a meta-synthesis is to synthesize the findings of qualitative studies and reinterpret the data;
On the other hand, when we need to examine both the quantitative and qualitative studies, we have to prefer a different method. However, what is the the correct option for us?
Is examining both the qualitative and quantitave studies at the same time a wrong approach?
Do we need to use meta-integration (aka mixed-meta synthesis) or systematic review? What are the differences between these two methods? What are the procedures that we need to follow?
Of course there are lots of studies in the literature but lets make it clear: which one is the "right" one? Let's share, discuss and try to make a consensus on this issue!
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Systematic review is defined differently by different institutes. For example, Cochrane has reserved this term for quantitative studies while Johanna Briggs Institute (JBI) uses the term for the synthesis of all kinds of studies. However, there seems to be a consensus that a systematic review has three characteristic features. First, a clear and well-defined research question. Second, a comprehensive literature search and selection of studies along with critical appraisal. Third, at least two independent reviewers must critically appraise the studies included in the review. The meta-analysis is a term reserved for quantitative studies and it is essential to perform a systematic review for meta-analysis.
With regard to your question, if it is wrong to examine both Qual and Quant studies, mixed methods systematic reviews are used. There are three types of mixed reviews. First, segregated (if you believe that both qualitative and quantitative approaches are distinct and should be analyzed separately). Second, contingent (if you believe that the qualitative and quantitative findings should be analyzed in a parallel manner and by moving back and forth), and finally integrated (if you believe that qualitative and quantitative approaches are not distinct, rather they qualify and complement each other).
Meta-integration or aggregation is the synthesis technique developed by JBI. It is based on the philosophy of pragmatism, that is, aggregation of studies in a way that they are more useful and practical for guiding practice and policymaking. This technique is mainly reserved for qualitative systematic reviews.
In addition, you may wana take a look at the 14 review typology developed by Grant and Booth to further clarify these terms.
References
Sandelowski, M., Voils, C. I., & Barroso, J. (2006). Defining and designing mixed research synthesis studies. Research in the Schools, 13(1), 29-40.
Grant, M. J., & Booth, A. (2009). A typology of reviews: an analysis of 14 review types and associated methodologies. Health Information & Libraries Journal, 26(2), 91-108.
Heyvaert, M., Hannes, K., & Onghena, P. (2017). Using mixed methods research synthesis for literature reviews: The mixed methods research synthesis approach. Thousand Oaks, California: Sage.
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I have some selected samples with basaltic in composition and vesicular texture. All of the primary constituents are fresh, and they don't have any alteration in thin sections. However, their vesicles typically filled by zeolite, carbonate, and other secondary minerals. I want to do a geochemical analysis for them. So I want to know, there are any effective methods to eliminate these secondary minerals, especially zeolites?
Looking forward to your valuable replies. Much appreciated!
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Dear Meysan Akbari,
I would dig out the amygdules manually under a good binocular. So you are in control what crystallized in the center or the rim. Then do XRD on the material. Simultaneously, prepare a thin section of the rock with amygdules which you can look at under the microscope, SEM or use for microprobe analysis to determine the chemical composition. Good luck!
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I would like to remark this to all are interested in publishing in Elsevier:
“European-Elsevier Scholarships”, \"Request for Papers\", \"Editorial/Reviewer Appointment\" & \"Manuscript Submission\"
- Fraudulent emails in circulation.
Please note: these are not Elsevier requests!
Cheers,
Stella
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Dear Maria Stella Ritorto , I have got the alert from Elsevier yesterday.
This is very good article with fine tips and suggestions.
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Hello,
I was doing a PCR for influenza viruses ( more specifically defective interfering segments) and i had good results beside 2 segments. I store them at -20 degrees and i was thinking to run them 5 cycles more ( initially all the sample were run for 15 cycles) . Do i need to make fresh samples or i can just reuse the 2 stored samples. I'm thinking just to place them in the PCR machine and add 5 more cycles to this 2 samples .
Will this work ?
I know the buffer, dNTPs , Taq polymerase and others are limited and have a specific life expectancy of PCR cycles, but 20 ( 15 already done + 5 which i will run )cycles from my presumption will not exhaust all the dNTPs and other reagents.
Thanks a lot ! Sorry for this stupid question !
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That likely refers to the concentrated stock, usually in 50% glycerol (so it doesn't freeze even at -20): enzymes last longer at cold temperatures, provided they don't actually freeze solid (ice crystals can do bad things to proteins).
Once you've diluted your Taq to make your PCR reaction mix up, it will freeze solid, and this will usually result in a stark drop in viability.
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What is Connolly surface method all about? I have found that It is mostly used to determine the surface volume of molecules.
How can this method be used to determine the size and volume of a nanocluster?Is there any software from which Connolly surface can be plotted and surface volume and size can be estimated?
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I can answer in Russian and would be added adout fractal
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I am trying to culture P. aeruginosa (PAO1) biofilms in 96-well plates. I have tried pipetting off the media, wicking it up with blotting paper or tapping it out. All of these methods results in severe disruption of the mucoid growth.
Can anyone advise on a successful method for removing the media, leaving just the biofilm?
Many thanks
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Another good way might be a microscope slide in a 50 mL centrifuge tube. If you don't mind going to the large volumes required for these (usually I use 25 mL). The other good thing about these is the ability to transfer the slide to a new tube with fresh media. This allows growth over multiple days/week without completely starving the biofilm. I do grow these with rocking (usually a shaker set to 100 rpm).
I actually have 50 of these tubes going right now with PAO1 and it is giving some really robust biofilms on the slides.
You can also try this method with the 24 well plate and use small plastic/glass cover slips. It seems to work but it depends on what you want to do with the biofilm or how high throughput you need it to be.
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If you build your own corpus to address specific research questions, which method to you use to make sure It is saturated? I'm interested in methods as I work on digital data and I wonder which method is more efficient and less time-consuming.
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In Corpus design, the "saturation corpus" is associated with the concept of "representativeness", developed by Douglas Biber: <http://otipl.philol.msu.ru/media/biber930.pdf>.
Here are some other sources from the University of Lancaster that might interest you:
... Methods: e.g. a short paper from the University of Birmingham :
4. A quantitative approach to corpus representativeness: <http://www.lexytrad.es/assets/cl24_0.pdf>
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I work in microbiology laboratory in a Flavors company, We conduct many microbiological tests to detect microbial hazardous such as E.coli, Coliform, Yeast & Mold and Salmonella.
Recently we received sample From LGC standards (used for proficiency testing )
In order to detect Salmonella we preformed the following procedure:
1. I Prepared 225 ml of buffered peptone then i added 25g of the sample to the buffered peptone.
2. Incubation at 37C° for 24 h.
3. 0.1 ml of sample in puffered peptone were added it to 10ml of Rapport Vasilidis (RV)
4. Incubation at 42 C° for 24 h.
5. Streaking plates of XLD and Brilliant Green (BG).
The results of XLD plates was strange because the grown colonies appears yellowish in color as in picture (B) not black colonies as it supposed to be.
Regarding BG results were also confusing because the media color has changed to pink but the colonies color is a bit different from typical salmonella colonies picture (A).
So to confirm this result I prepared another Fresh XLD and then I preformed streaking on XLD plates from (BG) and from (XLD) plates which used first time to detect salmonella.
The result in the end confirmed that the sample contains salmonella as in picture (C) !
So I have a situation here because XLD and BG firstly (A&B) didn't give me the confirmation that the sample contains salmonella due to the difference in morphological characteristics, but after streaking on another XLD it's confirmed that the salmonella is percent.
I think that's may happened due to decomposition of some medium components because we let the medium on hot plate till boiling = (over heating)
Is this thinking reasonable or it may be another reason for this issue ??
If you have any informations regarding this issue please share it with me.
Many thanks in advance for your support... 🙂🙂
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I agree with your suggestion that over heating of the XLD media on hot plate led to break down of the media components particularly , Ferric Ammonium Citrate and thiosulfate which losses there action and not transformed to give black center color of Salmonella colonies that metabolise thiosulfate to produce hydrogen sulfide. The media should be heated without bioling perferably in a waterbath.
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I am struggling to find any good summer/fall/winter research school on mixed methods research (for business and management, the area of broader social sciences research - also OK). Ideally, it should be within UK and EU institutions/universities. Do you have any ideas/suggestions? Thank you!
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I think this summer school could be interesting: Mixed Methods Research.
It is a summer school provided by ESSEX SUMMER SCHOOL in Social Science Data Analysis (UK).
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Making CLEAs. Hi all this is my first time on the forum so not sure how this works.
sorry this is so longwinded:
I am working on a project comparing the reactivity of L-aminoacylase with a HIS tag, at different purification stages, and in immobilised (CLEAs) or free enzyme form.
I have had problems trying to immobilise the enzyme. (I have followed a procedure from Toogood 2001 for making CLEAs which was adapted from Cao 2000).
This involved ammonium sulphate precipitation (I used 80% concentration) of the solution containing aminoacylase. It is stirred overnight at 4°C, followed by slowly adding 1% (v / v) formaldehyde and leaving under the same conditions for 2 hours. The subsequent washing step on filter paper from the protocol would never yield any protein, so I now centrifuge the precipitant first to get a pellet, I then I wash and filter the pellet and dry it in a desiccator.
However the majority of the pellet is again washed away, which leaves me with concerns that the protein has not been properly cross-linked.
I only have One week left of my project to try and get good CLEAs. Has anyone come across any similar problems?
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9 years late, but many thanks Kai
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Being an academic of finance and accounting subjects, I always look for new and contemporary ideas, thoughts, research, methods, models, processes involved with the research in the broad area of finance and accounting. once I found a website containing researches in the last 10 years, but unfortunately I lost it in the bookmarks.
Can we share the sources for getting such resources for learning and enrichment of knowledge in Finance and Accounting?
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In view of the above, Business Intelligence applications built by specialized IT companies may facilitate conducting accounting and financial reporting of enterprises and auditing financial statements, and thus diagnosing financial fraud, falsified financial data and discovery of creative accounting because the development of analytical platforms is determined by the progress of information technology making it possible it is an objective, multi-criteria analysis of large data sets.
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Recently, I got problems about Pichia expression. My target gene was inserted into pPIC9K vector via EcoRI and NotI restriction sites. And then the SacI digested construct was transformed into GS115 by electroporation. More than 200 transformants were obtained from the MD plate, and whcih was collected and subsequently plated on YPD plated with G418 gradient(0.5mg/ml to 3mg/ml). Only a few colony formed on the high G418 concentration plate, and then the colony was streaked on new G418 plate for isolation single clone. Then the genome DNA was extracted and PCR was performed.
PCR employed 5'AOX1 and 3'AOX1 primers gave 2.1Kbp bond, indicated the AOX1 gene from Pichia was still remained. however, PCR with my target gene speical primers, no bond was obtained. And subsequent western blot analysis shown that no target protein was expressed by induced expression.
Help!!!Where am I going wrong?
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I had the same problem.. on transformation into pichia followed by PCR initially I got 2 bands (one corresponding to 2.2 and another my gene of interest). But after few days I repeated the PCR since I was facing issues with expression, and to my surprise this time I got only one band corresponding to 2.2kb... My gene was lost from pichia. How do I deal with this problem?
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When should I change the water including antibiotics cocktail for antibiotics-induced microbiota depleted mice (AIMD mice)?
Hello, fella researchers.
I am trying to make AIMD mice for my study.
I have seen many papers to find method.
The antibiotics cocktail including ampicillin (1 g/L), neomycin sulfate (1 g/L), metronidazole (1 g/L) and vancomycin (500 mg/L) will be dissolved into the sterile drink water and provided to mice for 4 weeks.
But, I can't find when should it be changed. Every once in two days? I can't find.
So, If there is anyone who knows this or did this before, please help me.
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Hi Carl. I first use the antibiotic cocktail:
• provided ampicillin (A; 1 g/L; Sigma), vancomycin (V; 500 mg/L; Abbott Labs), neomycin sulfate (N; 1 g/L; Pharmacia/ Upjohn), and metronidazole (M; 1 g/L; Sidmack Labs) in drinking water for four weeks(Wang et al,2015)
However, the ampicillin has strange smelling and mice unwilling to drink it. I then only use remaining 3 antibiotic but the mice still don't want to drink and dehydrated in 5 days. Maybe gavage is a more efficient way.
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I just want the name of the methods....
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If you fix it with %30 formalin it is not possible...
All best
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I am trying to revive my cells and I don't know what they're being affected by. I have thoroughly cleaned every corner of the culture room and incubator; done all possible cleaning practices to avoid all possible sources of contamination. Now my cells are not adhering to the surfaces of the plates. They become rounded and are floating in the media. What additional techniques can I try to establish my culture? I don't know whether it is some biological contamination or some physical factor that is playing a major role.
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Were you ever able to resolve this problem? I am seeing the exact same phenomenon for over 2 months now and after going through all of the same things you did, I am at a loss as to how to resolve this. Any insights would be greatly appreciated!
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What research techniques and methods do you most often use in your research work, in scientific research?
Please answer
Best wishes
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The analytical techniques that I use are optical analysis Scanning Electron Microscope/Energy Dispersive X-ray Spectroscopy (SEM-EDX), inductively coupled plasma-mass spectrometry (ICP-MS), Fluorescence Spectrophotometry, Rock-Eval pyrolysis, Gas Chromatography-Mass Spectrometry (GC-MS) and TIMS-Phoenix Mass Spectrometry.
Best Regards Dariusz Prokopowicz
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I'm doing research on fault detection ,classification and identifying ,I have found out fault detection using  wavelet analysis need a guide about other methods.   
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I don't want to ask so many questions and I think they are not that one-of-a-kind material, how can I look for specific problems in ResearchGate?
cheers
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Dear Wenzel Czepluch,
Currently, it is not possible to conduct advanced search of publications, authors, .. on the Research Gate portal. Many users of the Research Gate portal indicate that it would be useful to enrich the publication search tool, authors, .. with additional, advanced options. Enriching search algorithms with additional options could be done in the same way as it does in Google search. Some similar solutions of advanced search functions already exist on some other social media portals.
Best wishes
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I am looking for methods of measuring of the Seebeck Coefficient or devices alternative to Netzsch SBA 458 Nemesis. It will be great if the simultanous  analysis of Seebeck Coefficient and electrical conductivity would be possible.
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There are significnt differences between Seebeck systems on the market (setup, robustness, accuracy). Netzsch lab performs demo measurements free of charge.
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Dear Community,
I am writing review articles in which I want to describe procedures from other papers.
Indeed, It is clear that the origin of data must be listed. But how detailed can a procedure from a foreign source be written down - in own words? Especially when specific procedure parameters (e.g. concentrations of chemical components, temperature etc.) are extracted from a paper - are there dangerous copyright traps i need to consider?
Any help is highly appreciated..
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Hi, the main thing is to indicate the source, and this does not mean that you should use the same words, but you must follow the basic meaning of the word.
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I am aiming for hyaluronic acid-based microgels particles, which are homogeneously modified with DNA. Therefore, SH-modified DNA, which is additionally functionalized with a fluorescence dye is couple to a PEG-maleimide crosslinker and mixed with hyaluronic acid to from W/O-droplet-emulsion. By following the gelation process using fluorescence microscopy, I observe diffusion of my DNA (around 1200 bp) to the W/O-interface giving particles with DNA only at the surface (like a corona). The only thing that I can think of is that this might be an issue of electrostatic repulsion between polyanionic hyaluronic acid and the DNA during the gelation process. Can this effect be avoided by adding additives to the emlsion, like CTAB or something else that lowers the repulsion? Or do you have any other reason, why I oberserve this and how to avoid it?
Thanks for your help !
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I wanna use hyaluronic acid-based hydrogel particles for gene expression on microscale. I know, hyaluronic acid is not the best candidate as a polyanion. There are two reasons why I am aiming for hyaluronic acid based platform. The Gels need to be porous enough to allow for diffusion of the ribosomes throughout the particles. Second, the HA is further functionalized with binding motives and therefore HA needs to be quantitatively functionalized (possible due to the carboxyl group of HA). So switching to chitosan or agarose is not really an option.
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Has anyone dealt with technical issues with both primary rat neuron/glia co cultures and human iPSC neuron astrocyte co cultures (both on PDL), where the cells seem to ball up near the cell body and cause long filament string like processes to one another? We think it could be due to the coating or a cell signaling process? See image comparison of healthy vs. "spindled" like cells. Apologies for the bad quality image due to uploading on here.
Any technical advice welcome. Thank you!
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Hello,
I routinely do primary cultures of hippocampal and cortical neurons. Sometimes, I I saw such similar neuron clumps in the dish. Here are two important things you might want to consider to solve this problem. 1) Number of cells and (2) PDL coating. You could think of reducing the number of cells to seed. And, also change your PDL stock solutions. In my case, PDL coating was the reason. I have changed the stock solution and also increased the working concentration of PDL for coating the dishes.
One more thing, do you observe neuron clumping only on glass coverslips or also on plastic coated dishes?