Thomas Hobbes. 1588-1679.
For words are wise men's counters,--they do but reckon by them; but they are the money of fools.
The Leviathan. Part i. Chap. iv.



Brian O'Rourke
Johns Hopkins University

-----Original Message-----
From:   Michael Ignatius [SMTP:[log in to unmask]]
Sent:   Thursday, November 05, 1998 4:27 AM
To:     [log in to unmask]
Subject:        Re: Autofluorescence reduction in mammalian cells?

Hi Cheryl,

First I hope that is the correct spelling for your name.  Secondly it was
great talking to you -you are a wise soul, a rarity in the sci world.  I
look forward to more conversations in the weeks to come, sprinkled in
around science discourse, of course.
Thirdly I came across this info last night and thought it might help.  It
came from the confocal group.  In the header you;ll find the address.  I
think you subscribe by email to the address and in the subject text say
subscribe, and then they will send back a confirming reply that you have
two days to acknowledge.

Hope that works, and hope the discourse below helps as well.

All the best,

Mike
PS the doctor visit was fun.  Makes it all more real.  We are proceeding
with things, though we are alarmed at the prospect of telling families.

>This is the best reply I saw in a similar discussion about two years ago:
>
>From: Martin Poot <[log in to unmask]>
>Reply-To: Confocal Microscopy List <[log in to unmask]>
>To: Multiple recipients of list CONFOCAL <[log in to unmask]>
>Subject: Autofluorescence still
>Newsgroups:   bit.listserv.confocal
>
>Since cellular autofluorescene seems to stir a continuous debate and was
>already an annoying aspect of life when I did my Ph.D at the Erasmus
>University Rotterdam (1989) here my two cents.
>
>We (and many others) found that autofluorescent material resulting from
>lipid peroxidation accumulated in the lysosomes.  You could overlook it as
>long as cells were dividing vigorously and they diluted out the stuff.  You
>could stop the formation of autofluorescent material by saturating your
>culture medium with vitamin E (Poot et al. (1985) Gerontology 31, 158-165).
>
>
>As to the chemistry of the autofluorescent material, you may like to look a
>Eldred et al. (1982) Science 216, 757-759.
>Chio and Tappel (1969) Biochemistry 8, 2821-2832.
>Esterbauer et al., (1987-1989) Biochemical Journal (sorry I lost that one).
>
>The upshot is that a reaction of an aldehyde (resulting from peroxidation of
>a polyunsaturated fatty acid (PUFA) with an amine (eg. an amino acid or a
>peptide)) leads to the formation of a fluorescent Schiff's base.  Depending
>on the size of the conjugated system created by this reaction, you will get
>fluorescent material of any color (see the TLC plates of Eldred et al.).
>
>At 09:29 AM 11/3/98 -0800, you wrote:
>>Satyajit Mayor wrote:
>>>
>>> Dear Users,
>>> I have a very specific question regarding reduction of
>>> autofluorescence
>>> from serum and medium components used for culturing mammalian cells.
>>>
>>> We have found that there is considerable autofluorescent material that
>>> fluoresceses in the red region (590+ nm), and this interferes with
>>> quantitation of fluorophors in this wavelength range whilst imaging
>>> mammalian cells in culture. (eg CHO cells).
>>>
>>> Some months ago I noticed a posting on the confocal list talking of
>>> using
>>> some kind of anti-oxidant to reduce autofluorescence in cells in
>>> culture.
>>> Does anyone remember the compound used and its mode of use?
>
>
>
>Larry D. Ackerman
>Lily & Yuh Nung Jan Laboratories
>Howard Hughes Medical Institute
>UCSF, Box 0725, Rm U226
>533 Parnassus Ave.
>San Francisco, CA 94143
>
>(415) 476-8751  FAX (415) 476-5774
>[log in to unmask]