Colleagues:
I received this query from a graduate student. If anyone has any
experience with sorting phospholipid vesicles as indicated below, I would
appreciate any comments you may have.
Thank you,
Julie
>Date: Thu, 19 Aug 1999 15:53:25 -0500 (CDT)
>From: william kevin meisner <wkmeisne@midway.uchicago.edu>
>X-Sender: wkmeisne@harper.uchicago.edu
>To: jauger@flowcity.bsd.uchicago.edu
>Subject: feasibility query
>
>Julie:
>
>I spoke with you briefly today about the feasibility of my prelim proposal
>and you suggested that I email you some specifics to post on the internet
>for comments.
>
>My goal is to study the dynamics of single enzyme molecules. A recent
>technological advance in phospholipid vessicle formation will enable me to
>trap enzymes in vessicles less than 5 micrometers in diameter. Ideally,
>each vessicle would contain a single enzyme molecule, and this could be
>accomplished if vessicle formation were carried out at very low enzyme
>concentration. Of course, the majority of the vessicles would be empty, so
>I must find a way to isolate those containing the enzyme.
>
>My hope is that this could be done with a fluorescent cell sorter, but I
>am woefully ignorant of this apparatus. Are single phospholipid
>membranes too fragile to successfully pass through the machine? Can
>vessicles of this size be accurately separated? Will tryptophan
>fluorescence (perhaps stimulated by a laser) from a single enzyme molecule
>be brilliant enough to enable detection? Would another method of
>detection be more robust?
>
>Fortunately, only a small number of vessicles are necessary for my
>proposed study, so their isolation need not be efficient.
>
>Any and all comments regarding the feasibility of this proposal, or other
>alternatives, will be received with gratitude.
>
>Thank you for your time.
>
>Kevin Meisner
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****
Julie A. Auger
Director, Immunology Applications Core Facility
University of Chicago
5841 S. Maryland Ave. MC-1089
Chicago, IL 60637
e-mail: jauger@flowcity.bsd.uchicago.edu
telephone: 773-702-9212 lab 773-702-9261 office
fax: 773-702-3701
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