Annexin PI staining
Hello everyone, I would like to know why only a few cells (around 30%) stain with annexin V while the rest of the cells are still not undergoing apoptosis post drug treatment. Has it got to do...
View ArticleWhat are the uses for positive and negative controls in flow cytometry?
Hello everyone, Sorry if this is a beginner's question or that it has been asked (and answered) before, but I would just like to know: what is the use of a positive control (eg:...
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Counting beads binding to RBCs
Hi guys,, Sooo...I have been using counting beads to quantify the concentration of RBCs in a mouse. However, when I count 1000 beads/sample for example, half of my bead population binds to my RBCs....
View ArticleHelp with the Red Blood Cell Lysis
Hello I am writing from Peru. Currently I am working in a Flow cytometry laboratory in oncohematology. The question is that we work with the Euroflow consensus and of course we work with manufacturers,...
View ArticleProtocol for human dendritic cell population in blood
Hi, I'm new to Flow cytometry and I'm looking for a protocol to stain and gate human dendritic cells from blood using flow cytometry. The Panel should incluce CD86, CD40 and maybe CD80, we have BD...
View ArticleSample is not running into the flow cytometer
Recently, the flow cytometer (BD FACSort) that I am using (with other labmates) is having some problems. The Drain, Fill and Backflush functions are working fine. However, the Run function is not...
View ArticleNo-wash fixation/permeabilization kit for RBC
Is there any no-wash kit that can be used for staining intracellular antigens in RBC? I need to fix the cells, permeabilize them and stain the intracellulat antigen, without washing. Beckman-Coulter...
View ArticleFacscan Repair. Peristaltic Pump connector?
I have acquired a Facscan, 1987 model. Of course, no warranty or service from auction. I found it is missing only one component which I managed to acquire. It's the whole assembly of Peristaltic...
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Question about FACS of maize protoplasts
Hi, I was wondering if anyone has experience on fluorescence-activated cell sorting (FACS) of plant protoplast. After PEG-transfection of the fluorescence probe into the maize protoplasts, I started to...
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Where to gate with overlapping population?
My fluorescent cells are only weakly fluorescent so they overlap with the non-fluorescent cells as in the below image. Would it be acceptable to move the gate to the left to get more of the fluorescent...
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FAC Sorting of live cells based on intracellular protein or transcript expressio
Hi Bioforumers, Has anyone successfully FAC sorted live cells based on their expression of a specific intracellular protein or gene transcript? I was considering labeling cells expressing my gene of...
View ArticleCorrect filter
I have to view a Alexa fluor 660 stained protein after intracellular staining, so i used the APC channel settings. Are they interchangeable ?
View Articleflow cytometry of intracellular protein staining
Hello , I am a doctoral student at the Faculty of Sciences Rabat Morocco, I am working on the flow cytometry analysis of an intracellular protein on different lymphocytes, For this purpose I make...
View ArticleIndustrial use of Cell Sorting Technology
Hello Everyone, I apologize if this is not the correct area for this but I felt that people in tune with flow cytometry may be able to lend me a hand. I am trying to compile an exhaustive list of...
View ArticleYogurt fermentation
Hello, I'm a High School student from Spain, and in the last year of high school we have to present a final project. My topic was yogurt fermentation and the study of the different techiques of...
View Articlenegative control readings
Hi. I would like to look into the effect of a compound on treated and untreated cells. For my negative controls (comprising of untreated cells), upon staining with antibody, I get a range of 1 to 2%...
View ArticleMulti-staining 7-aad, Annexin-V, Cell Trace CFSE, Dyecycle Violet
Hi, I had a question for the Flow experts here. Has anyone tried staining with multiple dyes - 7-AAD, Annexin-V (PE) - part of Annexin V Apoptosis Detection Kit from BD Pharmingen. These are to be...
View ArticleFixation for surface epitope detection
Hi, Is it advisable to not fix the cells after extracellular staining for surface marker using flow cyto? I mean, if I will directly go for detection after staining of the samples. Not planning to...
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Two distinct populations observed on a homogenous cells
Hi all, I often get two populations from my results of electroporated cells compared to non-electroporated control (pic attached). Left is non-electroporated,, right is electroporated....
View ArticleIsotype control vs. stained control
Hi, For flow cytometry I used stained negative control (cell not expressing the surface protein but are stained with antibodies similar to sample) and there are no background noticed for all 3...
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