Technique to distinguish eukaryotic and prokaryotic cells

[golfyfan]

Hi guys, this is my very first post, so if this is the wrong section, please move/ remove!

I've been searching for a quantitative assay that will allow me to quantify the number of eukaryotic cells in a gel along with bacteria infected. The eukaryotic cells are going to be fibroblasts and bacteria will be e. coli, pseudomonas aeruginosa, and staph aureus.

I'm hoping I won't have to do staining/ sectioning if I can avoid it. So something similar to an MTT assay, an enzymatic assay that works on eukaryotic cells and not on prokaryotic cells would be fantastic!

Any suggestions would be greatly appreciated!

[Tigerlilly]

Hey there,

I have a quiet similar problem. I have been searching for several techniques and have tried different approaches. Did you find an appropriate protocoll to separate bacterial and human DNA? I would be very pleased if you could give me some hints or publications that my help me to solve the problem.

Many thanks in advance!

[mmorgan]

How about quantitative PCR?

[Tigerlilly]

Yes, thats the step to determine how much is in there. But we want to enrich the bacteria first and reduce the eukaryotic content. I am thinking of a cesium chloride density gradient but would be greatful for advice.

[martinabbi]

hi everyone,, really desperate to know the solution of this ....

[Grossbouff]

Interesting problem.
Depending on the eucaryotic cell line, there may be a possibility to quantify them via a specific marker. For example, MCF-7 cells can be quantified by the CEA marker, I have tried a kit and it works.
As for the prokaryotes, I would simply grow an aliquot on agar plates. This way you quantify only live cells, of course.