[Histonet] Double fluorescent stains for apoptosis

[Chan Wai Kam]

Hi Kelly,

Thanks for your understanding.  Frankly I did feel chastened and thought =
I'd better go and do my homework more thoroughly.  It's true that I had =
hoped that some kind souls in histoland could offer me their advice on =
the correct protocols or kits to use as the kits are so costly and I'm =
wandering into unfamiliar territory. =20

I'm not a clinical but lab staff in research and my work has been mostly =
on routine histology of decalcified tissues.  I've no prior experience =
in apoptosis and so find it a bit confusing with all the protocols I've =
found on the web.  The PI whose project I'm assisting isn't of much help =
either as he was on a short attachment with our dept, finished the =
experimental surgery part and left the bulk of the work for the lab =
staff to complete.  He's happily in some other country and would drop an =
email every so often expecting results.  So frankly I'm unsupervised and =
trying to do the job as best as I can. =20

The Roche kit I used (In Situ Cell Death Detection Kit, TMR red for =
fluorescent microscopy analysis) comes with a protocol but the results =
do not seem accurate as there were too many cells stained positive.  I =
also realized too late that I should be going for something that shows =
up the unaffected cells as well since we need to get a percentage.  So =
back to the web I went to do more searches but still not any wiser.  And =
that's when I thought about all those experts out there in histoland who =
could perhaps point me in the right direction. =20

Thanks again,
Wai Kam

 =20

-----Original Message-----
From: kelly.mcqueeney <@t> bms.com [mailto:kelly.mcqueeney <@t> bms.com]=20
Sent: Wednesday, May 18, 2005 9:21 PM
To: John Kiernan
Cc: Chan Wai Kam; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Double fluorescent stains for apoptosis

The reason people subscribe to histonet is to reduce time spent "going=20
to the library and spending half a week immersing yourself in the=20
published literature of cell death and the methods used to recognize how =

it happens." This is achieved by asking your peers and "experts" for=20
help. You should copy and send this response to everyone asking advice=20
when they can simply take a week and look it up. I know I don't have=20
that kind of time, and it is clear that Chan Wai Kam does not either.

Chan Wai Kam:  I would help you, my friend, but I do not have experience =

with the Roche kit.

Kelly McQueeney
Research Scientist
Bristol-Myers Squibb

John Kiernan wrote:

>Dear Chan Wai Kam,
>
>Your enquiry clearly indicates that you don't know how
>your bought kit works. The TUNEL (not tunnel! It's an
>acronym for Terminal Uridine Nick-End Labelling) family=20
>of methods has many shortcomings, all thoroughly=20
>documented in peer-reviewed papers that are cited in
>textbooks. TUNEL, intelligently used, has its place,=20
>alongside other techniques that indicate modes of
>cell death.=20
>
>Your "we need a protocol" plea is a way of saying=20
>"Tell me exactly what to do" instead of going to the
>library and spending half a week immersing yourself
>in the published literature of cell death and the
>methods used to recognize how it happens.=20
>
>Are you a clinical resident required to do a research
>project in a few weeks? This is the impression that
>I get from your email. The only sensible reply is
>"Go to the library and tell your boss to go there=20
>too".
> =20
>

--=20
No virus found in this incoming message.
Checked by AVG Anti-Virus.
Version: 7.0.308 / Virus Database: 266.11.12 - Release Date: 5/17/2005
=20

--=20
No virus found in this outgoing message.
Checked by AVG Anti-Virus.
Version: 7.0.322 / Virus Database: 266.11.12 - Release Date: 5/17/2005
=20