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bacterial viability assay


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4 replies to this topic

#1 OKSO

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Posted 05 August 2009 - 07:37 AM

Hi,

I am going to look at bacterial viability after ingestion by macrophages. I will lyse the cells and initially plate out the lysates to count cfu's
However, I need to do this at a high thoroughput level, and therefore would like to use a fluorescent/colormetric assay in 96 well plates to determine bacterial viability.
I have heard conflicting results regarding the alamar blue (resazurin) assay, and wonder if anyone has used similar assays? or even looked at fluorescent assays such as calcein?
Thanks!

#2 genehunter

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Posted 05 August 2009 - 09:15 AM

You can look into some DNA binding dye on fixed bacteria.

#3 GeorgeWolff

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Posted 06 August 2009 - 01:50 AM

How do you propose to validate whatever technique you emply in this application?

#4 OKSO

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Posted 06 August 2009 - 02:04 AM

I guess that's why I'm using this forum.

An obvious method would be correlation with cfu's, but am open to whatever has been tried and tested by others...

#5 pDNA

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Posted 15 November 2009 - 01:36 AM

you can use for example SYTO-9 (live) and propidium iodide (dead) ...but i don't know how this interferse with your macrophages. I don't get it how you would do this in 96-well plates? How will you do the read out of such an assay? Maybe you can comment on that?

Regards,
p




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