Request

From: Manoj Tadem (tadem_tadem@yahoo.com)
Date: Wed Nov 21 2001 - 12:10:47 EST


Hi there,
 I am Manoj Tadem doing PhD in United Kingdom. I am
trying to establish a protocol for staining of DNA
using Hoechst 33342. I am very new to Flow Cytometry
and am short of an expert advice regarding this
particular protocol.
Its an Epics Elite Coulter Flow cytometer. Till now I
have tried about three times, but could not establish
a reliable staining procedure.
I have few questions regarding this protocol.

1)I have used PMT1 and PMT2 for detecting the
flouresence of Hoechst 33342. Is it the correct way?
If not kindly suggest me the appropriate PMT's for
detection of Hoechst Dye.

2)Which value in the statistics, should be taken as
measure for flouresence intensity?
For instance, is the X mean or area under curve or the
%value.

Could anyone kindly help me establishing a protocol.
For which I shall be grateful to the person.

Thanking you,

Your's truly,
Manoj tadem


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