SYBR-14, continued..

From: Michal Bochenek (mbochen@izoo.krakow.pl)
Date: Fri Oct 03 1997 - 00:13:50 EST


Hi, 
Thank you for responses,

On Tue, 30 Sep 1997, Tom Frey wrote:
> If you mean SYTO-14, see.. 
I mean SYBR-14, the component of sperm viability kit (Molecular Probes), 
the second of the components is PI.

> ...if you have PI around the dead cells are going to show some quenching 
>due to energy transfer.
Tom is right: SYBR14 used along with PI gives energy transfer (SYBR14->PI)
in the 'dead' PI positive cells.
But the other question exists. Using SYBR14 alone and with time as 
a parameter I can see that after ab. 3-4 minutes of acquisition 
fluorescence of cells (spermatozoa) divides into two 'branches' - the 1st 
one remains on the same level while the 2nd one gradually drops down. 
This phenomenon repeats in the succeeding loadings of the *same* sample.
What is a cause of ssuch behaviour?

Thanks in advance,
Michal

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                       |  National Research Institute of Animal Production
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