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3
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1answer
33 views

Flow cytometric counting of apoptotic adhering cells

I have been trying to measure rates of apoptosis in epithelial cells by marking apoptotic cells with annexin V conjugated with phycoerythrin (PE) and sorting using flow cytometry. Unfortunately, the ...
0
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0answers
41 views

What is C-value for polyploid species?

I am curious to confront genome assembly sizes with flow cytometry estimates of couple of strange polyploid genomes, but I got really confused about meaning of C-value. According verbal definition ...
1
vote
1answer
30 views

Flow Cytometry Channels

I did a flow cytometry experiment for the first time. I had three conditions: (1) unstained healthy monocytes (2) healthy monocytes stained with green/red (488/570) from the Live/Dead thermofisher kit ...
0
votes
1answer
48 views

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

In a nutshell, is it possible to FACS a sample that is in oil rather than in a buffer (water based solution)? I would greatly appreciate any help with this. Thank you
1
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0answers
17 views

What enzymes I shall use for preparing single cell suspension from cardiac tissue for flow cytometry?

Can anyone suggest which enzymes I shall use for preparing single cell suspension from cardiac tissue. I will be looking at both cell surface and intracellular antigens involving cardiomyocytes and ...
1
vote
1answer
132 views

Flow Cytometry to learn about Cell cycle

I am posting my Flow Cytometry data here again because I realize that there are many people who are willing to help me, and I can learn a lot from you guys. In this cell culture Lab, we use Cell U-...
1
vote
1answer
402 views

Flow cytometry data Analysis [closed]

everyone. Since I am new in the flow cytometry technique, I am not really sure how to analyze the result. We use cell U-937 Lymphoblast.All Cells Could Anyone help to explain me this one? Thanks ...
4
votes
2answers
383 views

Cell dye in red fluorescence spectrum

There are some cell dyes available for stianing cells intracellularly / membrane. I am aware of other channels (CFSE \ PKH26). But I want to stain cells for flow cytometry with a 'red' dye. I have ...
2
votes
0answers
57 views

Flow cytometry analysis

I've performed an experiment wherein I have stimulated cells (cell line) with a drug across different time-points - in its unmodified (Drug) or modified form (H-Drug or M-Drug). All cells are seeded ...
3
votes
1answer
215 views

Purpose of antibody wash

It is common practice after surface staining cells for flow cytometry analysis to wash the antibody out of solution before analyzing a sample. I have tried analysis with and without washing the ...
0
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0answers
66 views

Differing Flow Cytometry Genome Sizes and Sexual Mating Implies Differing Chromosome Counts

Do the fundamental principles of genetics strongly suggest the following is a derived conjecture? If 2 flowering plant species A and B have widely differing genome size (at a minimum a 50% difference)...
2
votes
1answer
2k views

Staining cells for FACS at 4 degrees or ambient temperature

I'm sorting a 293 derived line. One thing that is worrisome for me is cell viability after the sort. Usually I have been staining and washing at room temperature (on the benchtop) on a nutator. I have ...
2
votes
0answers
105 views

Can flow cytometric analysis differentiate between a parent contributing longer chromosomes versus multiple smaller chromosomes?

For flowering plants, can flow cytometry of seed from muti-generational interaction of two species and their offspring help us decide if wide differences in genome size for the two species and their ...
0
votes
0answers
529 views

Jurkat single cell clonal expansion

I'm hoping to do a clonal expansion of Jurkat, expressing my construct which is currently transduced virally. From the FACS data I currently have roughly 30-40% expression in my culture but I need it ...
7
votes
1answer
4k views

Magnetic-activated cell sorting vs. FACS

When sorting cell populations it is possible to use either magnetic-activated cell sorting or fluorescence-activated cell-sorting (FACS). I am wondering when you would choose either technique and what ...
3
votes
2answers
2k views

Alternatives to commercially available flow cytometric analysis software for use in cell cycle analysis?

I know that many labs use either FlowJo or FCS Express to plot and analyze their flow data. These seem like the two standards for most labs (and I have used FlowJo before). However, my current funding ...
3
votes
0answers
542 views

Accuracy of genome size estimation by flow cytometry

I'm working on a genome project and using an in silico k-mer analysis to estimate the size of our genome based on the available Illumina reads. The k-mer based estimate is consistent across a wide ...
5
votes
2answers
93 views

Flow cytometry issues

I'm having problems with data analysis here. I have flow cytometry data being collected on a Fortessa, and when I import them into FlowJo 8.7, all of my fluorescence values are systematically 10X ...
5
votes
2answers
1k views

Isotype control antibodies in Flow Cytometry

In a Flow Cytrometry, one can add an Isotype Control Antibody to allow another antibody to bind more specific to the cells. My question is, how can the Isotype Control Antibody add specificity to the ...
13
votes
3answers
42k views

Hoechst or DAPI for nuclear staining?

When is it best to use Hoechst vs. DAPI for nuclear staining? They seem to be very similar on paper. Are there situations where one is clearly preferable?