Re: Advice on Cell Sorter

From: Matthew Wikstrom (mattew@cyllene.uwa.edu.au)
Date: Fri Aug 20 1999 - 01:28:00 EST


Hello Stan, and the rest of the group.

I have been in charge of a newly formed core facility for the last twelve
months that has two benchtops (a Scan and a 4-colour Calibur) and a
Vantage (3 laser, 5 colours).  I was hired as a part-time operator and
manager for the facility after the instruments had been acquired.

I think all your questions can be answered by addressing a single issue,
and it relates to the prospective users of the facility.  What is the
level of flow work and what kind of applications have to be provided
for?  It might be difficult to accurately the second part of the
question, but you should have some idea what everyone wants to use it
for.  By answering these questions, you will get exactly what you need,
and hopefully have some money left over to acquire a good operator and
some new reagents.

The Vantage that was purchased for the facility here was spec'ed up to
provide for some of the most sophisticated applications, and included
TURBOSort.  As an experienced 4-colour user, I couldn't wait to start
powering all three lasers for 5-colour work, but there was a problem.  No
one in the area (Perth, Western Australia) was ready to do 5-colour
experiments.  Few do 3-colours.  The reason for this is simple--this is
an area where there are few flow cytometers, and as such, flow is not
widely used.  It was the perfect place to establish a core facility, in
terms of providing a worthwhile service, but, no instruments equals no
experience equals no motivation.  Put simply, I've got a Porsche, but no
where to race it.

It's true, in time and with some measure of vigorous encouragement, the
level of use and the sophistication of experiments will increase as the
local workers get "used" to the technology, but it will be a matter of
years, not months.  By then the potential of the Vantage will start to be
realised, but what of the money that was initially spent to spec it up?
It might have been better spent in anticipation of service contracts (as
bought, the Vantage came with a 3 year warranty/service contract), which
will come to nearly AUD20,000 to cover all three in two years time.  Or
it could have been spent on reagents to get new users started or to help
current users do more sophisticated experiments.  I don't know what the
cost differences were, between a single-laser, 3-colour Vantage without
TURBOSort and the fully-spec'd (Porsche) version, and I'm sure it was an
attractive offer, but the Porsche would have cost more.  If you don't
know if you'll be able to get more money down the track to upgrade the
instrument, then get all you can now, because users will always be
looking to do more once they get started.

So, after a year of "trading", let me tell you about the success of my
facility.  I've been able to do a bit of sorting, but the instrument has
not been turned on more than once a week, and that has included my
training.  There is interest in it, for sure, but these things take
time.  I have yet to fit the TURBOSort, it's still in the cupboard, and
while it would have been nice to dial up the sort speed on one or two
occasions, it didn't stop me from sorting the sample.  I will fit it
shortly, but only in anticipation of it's use (and because I like to
drive fast cars, even if it is only to the corner store).  I haven't had
any problems with the Vantage at all, have not been frustrated by any
aspect of it, but then, I've haven't had any reason to explore its
limitations.  As far as safety goes, the Vantage comes fully equipped
with covers and shields to meet safety regulations in terms of laser
safety and sample safety.  Of course, live human samples should be
inactivated with fixative etc, and/or appropriate inoculations made for
the operator and research worker.

The expertise of the operator should match the level of work that is
going on in your area.  Someone like Mario "Rainbow" Roederer would
wither and die without multi-colour users, while a novice would be
reduced to tears in the reverse situation.  Define your needs, and then
you will be able to meet them.

On the issue of service contracts, I believe in them.  They protect the
investment that has been in the instruments, and as the backbone of a
core facility, the instruments cannot afford to lay idle for any longer
than is absolutely necessary.

The only other issue are the unknowns.  What if you don't know what sort
of applications to provide for, or what if you want to have enough to
give the facility a broad appeal?  At the end of the day, you won't want
to turn away a customer because you weren't able to do the experiment for
them.  The only advice I can offer is to stick to what you know and do it
really well.  Rather than looking at the possibilities, consider the
realities of the work that is going on and where it is likely to lead.
I'm sure you will be able to find the money when you need it to extend
the capabilities of the instrument.

Good luck

Matthew Wikstrom
Lotteries Flow Cytometry Unit
Department of Microbiology
University of Western Australia
Perth, Western Australia
Email: mattew@cyllene.uwa.edu.au
Web: http://www.microbiol.uwa.edu.au/~MATTW/





"Ress, S, Stan, Dr" wrote:

> Advice would be greatly valued from those with experience in
> cell sorting, with regards to setting up a core, central facility. It
> is clear that a dedicated operator is essential, together with high
> speed sorting capability, or else the instrument will not be used.
> This of course can get quite costly. The sorter will likely be used
> by multiple departments for both experimental animal research, and
> applied clinical research including hematology and infectious
> diseases, cancer and molecular biology. The FACS Vantage Dual Laser
> bench sorter would be under consideration, but please note that I am
> not looking for pros and cons of different company products, but
> rather for general advice. Please could I have input on the following
> points:
>
> 1. Minimum basic configuration vs optional extras.
>     Foe example, is it essential to have Argon lasers, Fl-4 Detector,
> Turbo sorter at the outset, for robust output, or can these be later
> additions (budget permitting)?
>
> 2. What protective measures are necessary for working with infectious
> material such as mycobacteria and HIV, in animal and human tissue.
> Are there protective covers available that need to be installed? Are
> there operator guidelines published (I seem to recall something from
> cytometry the past year)?
>
> 3. Any references to recent review articles on sorting and its
> applications in biology, including molecular biology (gene
> applications), would be valuable.
>
> 4. Maintence contracts and insurance of lasers.
>
> I know this type of question has been posted before but there were
> few replies that I could find, as the requests were usually for
> direct replies off the list. I am happy to repect confidentiality in
> the latter case, but imagine the topic would also be of general
> interest, and as technology advances, needs to be aired periodically.
>
> I would be grateful for any advice.
>
> Thanks.
>
> Stan.



This archive was generated by hypermail 2b29 : Wed Apr 03 2002 - 11:53:52 EST