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Optimization of the flow cytometry

optimize cell instrument marker receptor

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3 replies to this topic

#1 H. Aly

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Posted 24 November 2014 - 12:38 PM

Hi 

I am new to flow cytometry, and I am going to use it to determine the expression of surface receptor. The flow in the lab was optimized to analyze blood cells not cell lines, so do I need to optimize/ change settings of the instrument to work for my cells?



#2 Tabaluga

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Posted 24 November 2014 - 01:27 PM

Run some unstained cells first and adjust the voltage settings so you can comfortably see the population in the FSC/SSC and differentiate it from the debris. Next, run a stained sample and adjust the voltage settings for the respective fluorescence.


Il dort. Quoique le sort fût pour lui bien étrange,
Il vivait. Il mourut quand il n'eut plus son ange;
La chose simplement d'elle-même arriva,
Comme la nuit se fait lorsque le jour s'en va.

 


#3 H. Aly

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Posted 24 November 2014 - 09:22 PM

Do I have to optimize it even if I am going to use the flow to detect the fluorescence only not the size of the cell ?

 

Does it has effect on the  surface receptor fluorescence detection ? 



#4 Tabaluga

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Posted 25 November 2014 - 05:52 AM

But you need cell size and granularity (FSC/SSC) to exclude debris, for example. And I'd adjust the fluorescence channel voltage so I can comfortably discriminate stained and unstained cells.

Il dort. Quoique le sort fût pour lui bien étrange,
Il vivait. Il mourut quand il n'eut plus son ange;
La chose simplement d'elle-même arriva,
Comme la nuit se fait lorsque le jour s'en va.

 






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