Non-replicating toxicity assay

Andy Pekosz pekosz at shy.neuro.upenn.edu
Fri Jun 21 06:37:49 EST 1996


In article <1996Jun19.052839.22761 at marlin.jcu.edu.au>,
nicholas.moody at jcu.edu.au (Mr Nicholas Moody) wrote:

> Hi, 
> 
> I am looking at the effect of an ultraviolet iradiated iridovirus on
> cell culture. The UV irradiated virus does not produce replicating CPE
> after inoculation, however, when the cells are inoculated at a
> relatively high MOI (equivalent to 1 - 5 infectious particles per
> cell) the cells lift of the culture flask.This effect is totally
> different to replicating CPE and is dose-dependent. Staining with 0.4%
> trypan blue (final concentration) to detect dead cells was not
> successful.
> 
> Does anyone know of an alternative to trypan blue staining or any
> other way I might be able to quantify this effect.
> 
> Thanks
> Nick Moody
> Biomedical and Tropical Veterinary Sciences
> James Cook University
> Townsville 4810
> AUSTRALIA
> 
> Tel: +61 077 81 4631
> Fax: +61 077 79 6371

A way to measure the metabolic activity of cells is with an assay called
MTT conversion.  MTT is converted from yellow to blue by cells which are
metabolically active.  It is very simple and will tell you if the cells
are being adversely affected in any way by your treatment.

It is a fairly common cell biology assay, here is the original and a more
recent reference.

J. Neurovirology 2:118-126, (1996).

J. Immunological Methods 65:55-63, (1983).

Good Luck!!

andyp.

-- 
Andrew Pekosz
Depts. of Neurology and Microbiology
University of Pennsylvania Medical Center
255 Clinical Research Bldg.
Philadelphia, PA 19104
email-pekosz at shy.neuro.upenn.edu
phone-(215) 898-3502
fax-  (215) 573-2029



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