Hi,
I have a problem that I hope you can help me with. I am doing in vitro
infections of primary macrophages with various live intracellular bacteria, and
I have great problems in finding a method to evaluate bacteria-induced cell
death of the macrophages / cell viability. To summarise:
- detachment of the adherent macrophages is not an option, which precludes
normal counting. Also, the cell growth and death is very uneven in the wells
and make microscopy methods difficult (biased selection of fields) - and tedious
- the presence of bacteria precludes methods like MTT, WST-1 etc. due to the
bacterial metabolism of these substrates. Similarly, crystal violet and neutral
red will also stain bacteria.
- LDH-type assays can be used in some experiments (short-term), but as I have
read that the stability of LDH in tissue cultures is about 9 hours, it will be
difficult to monitor cell viability or cell death during a 10-day infection
period?? Or can I??
I'll be very happy if anyone could help me out here,
Sincere thanks,
Trude
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