3 votes

Flow Cytometry Channels

I'll amend this answer as I get details from you. What is your cytometer, and how are the optics set up? You typically have 1-5 lasers: UV (355nm), violet (405nm), blue (488nm), green/yellow (~561nm) ...
CKM's user avatar
  • 8,109
2 votes

How are single cells sorted in an scRNA-Seq protocol by FACS?

we don't know the baseline of CD markers' expression levels yet? There are standard settings that can be used between runs that will give consistent results from one sample to the next. There are ...
iayork's user avatar
  • 14.2k
2 votes

How are single cells sorted in an scRNA-Seq protocol by FACS?

I will append the previous sentence in your protocol, and put what might help you in bold: To obtain B cells, NK cells and monocytes, a splenocyte suspension was stained with PE-Cy7-...
S Pr's user avatar
  • 6,222
2 votes
Accepted

Cell dye in red fluorescence spectrum

I've used the ThermoFisher brand of CellTrace dyes. Specifically, I co-cultured two populations of cells, one with CellTrace Far Red in the APC channel and another with CellTrace Yellow in the PE ...
CKM's user avatar
  • 8,109
1 vote

Cell dye in red fluorescence spectrum

There is a number of different dyes possible, as you can see in this figure (from here): A good overview can be found in the "Selecting Reagents for Multicolor Flow Cytometry" application note ...
Chris's user avatar
  • 51.6k
1 vote
Accepted

Concentrating cell concentration after subsampling

So the solution for this can be in two ways: calculate the total number of events (evenst/uL * total volume of sample at time X) and then devide by the initial volume. or perform a correction factor,...
Franco Grosso's user avatar
1 vote

Number of membrane proteins of a particular type per cell

If you know the stoichiometry of a particular ion channel and do something like patch-clamp, where you know the surface area being clamped, then you can measure the ion-flux per unit time and from ...
bob1's user avatar
  • 12.1k
1 vote

Can I use fluorophore conjugated antibody as neutralizing antibody?

In short, yes, most likely. When an antibody binds to the antigen at a region (epitope) that is needed for the antigen's activity then the activity is reduced ("neutralized"). The (proper) ...
Bob Tomas's user avatar
1 vote

CRISPR-Cas9 knockout

To answer your question multiple parameters which one should be taken into account. There are two main sets of parameters: gene specific and cell line specific. Parameters related to your specific ...
Dr. H. Lecter's user avatar
1 vote
Accepted

Non-nucleated cell-like population with RNA

Maybe you are seeing extracellular vesicles1? At least might be worth checking out since extracellular vesicles containing RNA have been reported within the hemolymph of mosquitos2. References: 1: ...
tyersome's user avatar
  • 5,598
1 vote

Flow cytometric counting of apoptotic adhering cells

Here some resources, Cell harvesting effects on Annexin V staining. Protocol for apoptotic analysis using Annexin V. Quite a long discussion about this issue. Another protocol. Cheers, Pedro
TumbiSapichu's user avatar
1 vote

Is it possible to sort droplets inside an oil phase with a FACS

It's possible that there's some specialized modified machine that can do it, but conventional flow cytometry machines can't do it. This article suggests one possible approach: Unfortunately, FACS ...
iayork's user avatar
  • 14.2k
1 vote
Accepted

Flow Cytometry to learn about Cell cycle

The basis of analysing flow cytometry data in regard to the cell cycle is the fact that DNA synthesis occurs in S-phase and therefore cells before S-Phase (G0/G1) have half the amount of DNA compared ...
Nicolai's user avatar
  • 4,391
1 vote
Accepted

Flow cytometry data Analysis

Disclaimer: Assumes you know how flow cytometry works. You've done a flow-based AnnexinV apoptosis assay here. Annexin A5 (AnnexinV) protein binds to phosphatidylserine (PS), which is typically in ...
CKM's user avatar
  • 8,109

Only top scored, non community-wiki answers of a minimum length are eligible