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Basic flow question


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#1 corduroy

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Posted 27 October 2009 - 07:04 AM

Hey all, I hope someone can give me a little guidance here.
Here's the scenario. Let's say I have 200k cells per ml in wells. Sample A is the control and Sample B is treated with a known substance that kills 90% of the cells at 48 hours. When I run flow at 48 hours, do I: use the same volume as the control to read or do I count out the same number of cells between the control and treated samples and read those samples (which would be 10x of the treated sample)? Readings would be done on PI or 7aad and several antibodies for expression.

I have a few thoughts but I keep on going back and forth on this and am unsure. If I'm not too clear, let me know - it's something I'm working on :) Thanks.

#2 canotto

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Posted 28 October 2009 - 06:43 AM

Hey all, I hope someone can give me a little guidance here.
Here's the scenario. Let's say I have 200k cells per ml in wells. Sample A is the control and Sample B is treated with a known substance that kills 90% of the cells at 48 hours. When I run flow at 48 hours, do I: use the same volume as the control to read or do I count out the same number of cells between the control and treated samples and read those samples (which would be 10x of the treated sample)? Readings would be done on PI or 7aad and several antibodies for expression.

I have a few thoughts but I keep on going back and forth on this and am unsure. If I'm not too clear, let me know - it's something I'm working on :) Thanks.


I think that you will find an answer to most of your questions regarding this issue in the following reference: King et al,Detection of dead cells and measurement of cell killing by flow cytometry, Journal of Immunological Methods, 2000




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