Staining

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Questions related to Staining

How do we handle vanishing or exploding gradients in deep learning neural networks ?

Techniques to handle vanishing or exploding gradients in deep learning neural networks ?

12 March 2023 1,600 3 View

How to choose the mitochondrial targeted sequences?

Hi guys, I want to transfer a protein into mitochondrion, therefore, i need a mitochondrial targeted sequence(MTS). Since there are lots of MTS, and I choose the COX8A, SOD2, and OTC. The first...

10 March 2023 4,905 0 View

Hi, I am willing to seed CaCo2 cells on 24 wells transwell - need a help with how to seed(how much) and more info. about the differntiation process?

how "much" cells I need to seed in a transwell and how to assure getting a monolayer?

10 March 2023 3,280 0 View

How to merge a protein sequence with its generated structure in PyMOL or any other software tool?

A protein structure was generated first using a deep-learning method, which only returns the visualized 3D structure, but not the sequence. Later, the sequence was generated by using another...

09 March 2023 9,182 10 View

BLAST search gives too high query cover percentages?

Hi there I am trying to design guide RNAs for cutting the yeast 2 micron plasmid using CRISPR. Using Chopchop, I got a 23 nt sequence which was suitable for gRNA design. When I blast the 23 nt...

06 March 2023 434 1 View

Which are the latest deep learning models for zero-shot image classification ?

06 March 2023 1,537 6 View

Does anyone have experience working with Wharton's jelly and menstrual blood mesenchymal stem cells (WJ-MSCs and Mens-MSCs)?

I am culturing menstrual blood msc and Wharton's jelly which do not grow well and die in part 4 and 5, especially Wharton's jelly does not grow well. While all the conditions are the same as...

04 March 2023 6,946 2 View

Can i check for cell apoptosis for 8 cell stage embryo?

i bought Elabscience®TUNEL In Situ Apoptosis Kit (HRP-DAB Method) for Tunel assay. Can this kit be staining the 8 cell stage embryos or only staining the tissue?

04 March 2023 2,934 2 View

Can any one provide me catalog number of LPS and interferen gamma for the stimulation of RAW 264.7 in terms of nitric oxide production?

can any one provide me catalog number of LPS and interferen gamma for the stimulation of RAW 264.7 in terms of nitric oxide production?

03 March 2023 7,972 0 View

How to improve RNA isolations for hard corals?

We are trying to isolate RNA from hard corals, however we are running into a few difficulties if anyone has advice. We have tried a couple of protocols.. one where we grind the skeleton and tissue...

02 March 2023 4,540 1 View

How to determine the cytotoxic effect of CAR T cells on lymphocyte B cell line?

28 February 2023 3,239 3 View

Western blot sample storage ??

I should do western blot experiment for detection of protein in liver sample .. i wash tissue with pbs then homogenize it with ripa lysis buffer and PMSF and Na3VO4 and then centrifuge it and...

28 February 2023 6,972 2 View

How to perform statistics on flow cytometry? What kind of statistics?

Hello, What type of statistical test would one use to find a significant difference in antigen expression? (Ex: Surface protein expressed in cancer cells vs. non-cancer cells) would one perform...

28 February 2023 2,188 4 View

How to avoid SH-SY5Y cluster formation in 96 well plate?

Hello All, I have been culturing SH-SY5Y cells and have observed that after plating cells (10,000/well) in 96 well plate, they start clustering by 48 hours (big countable clusters). I am not sure...

28 February 2023 6,394 2 View

Could someone recommend a software to reconstruct haplotypes from heterozygous sequences?

I have simple sanger sequencing aligned data with SNPs of about 80 specimens. How can I reconstruct the most likely haplotypes from them?

28 February 2023 1,677 1 View

How to inject beads/organoids into microfluidic devices with high loading yields?

I use the typical syringe pumps to load organoids (200-300 um in diameter) into my device; however, I don't see them in the chip! I swapped them with beads (similar size) instead and observed that...

27 February 2023 951 2 View

Anyone working with zebrafish astrocytes, microglia and/or pericytes culture?

Hello, I'd like to start zebrafish astrocytes, microglia and pericytes culture but I can't really find which are the best conditions and protocol to do that. Also, I may not have the possibility...

24 February 2023 2,116 0 View

Is there any established protocols for Adult Zebrafish H&E staining?I am particularly interested in knowing how to fix fish and for how many days?

My advisor provided me with these protocols. But I am not sure about the protocols which involve so many chemicals. https://zebrafish.org/wiki/health/disease_manual/recipes_and_protocols

23 February 2023 6,261 1 View

What implemented approaches are available to enable people using an online service to save their progress and return later to complete the service?

I'm working on a government form-building service to help public servants quickly and easily get their forms online. I know many folks in different jurisdictions are working on this same problem....

23 February 2023 8,123 0 View

Protocol to lyse cells inside hydrogels?

I am trying to extract DNA from a starting bacterial culture used for In-vessel composting which came in the hydrogel. I am not sure of the exact type of hydrogel, I tried sonication after...

22 February 2023 3,388 0 View

Does anyone know whether the antibody BG4 against G quadruplexes bind both DNA and RNA G quadruplexes?

In the attached paper (by Shankar Balasubramaniam's group) they have used the BG4 antibody to pull down G4 RNA. Does this mean the antibody binds both DNA and RNA, only one with less affinity? Can...

22 February 2023 7,520 3 View

Accuracy in Flurogold cell quantification in mice?

I have mice brain alices with FG staining without the use of anti-FG antibody. I have some very brightly stained neurons while quite a few somewhat faded cells in the background. Should i count...

22 February 2023 1,830 1 View

What is the most reliable method for quantification of RNA; Qubit, Nanodrop or Bioanalyzer?

For the same sample these give different values so which one to trust?

21 February 2023 7,651 3 View

Does T-bet expression differ in Th1 and Th0 cells? What is the best protocol for polarizing Naïve CD4 T cells into Th1 cells?

I have isolated Naïve CD4 T cells from mouse spleen and tried to polarize them into Th1 cells. For doing that, I have coated a 48-well plate with 1 µg/ml Anti-CD3, 100 µl each well, incubated at...

18 February 2023 3,241 0 View

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