RE: The automated cytometer is coming !!!

From: Gerhard Nebe-von-Caron (Gerhard.Nebe-von-Caron@Unilever.com)
Date: Fri Aug 03 2001 - 09:48:12 EST


Cytometers are fairly automated, that is if you buy al the necessary
workstations and the autoloader. In fact the standard 32-tube autoloader on the
Beckman Coulter XL has so far given us sufficient and extremely reliably walk
away automation for analysing experiments.

The 96 well format is not the default carrier as a lot of assays like in our
case are whole blood assays and we need to wash and lyse in volumes bigger than
what a standard 96 well plate takes. There are deep well plates that take ml
volumes, but using appropriate racks from Nalgene you can at least use 4 of the
eight channel (repeat) pipettes to either transfer samples from multiwell
plates or to add reagents in an efficient way.

I have tried to play around with 96 well filter plates. They are great with
bacteria, but I did not have much luck with mammalian cells . They do not like
to get dry which is required for efficient washing steps. However, as we can
now start to measure nine parameters at 50,000 cells per second in the Cyan we
might be able to do it all in whole blood w/o lyse. This would also abolish the
lysis artefacts and one could always capture unbound antibody to avoid a wash.


Regards

Gerhard


-----Original Message-----
From:	Akos_Szilvasi@biogen.com [SMTP:Akos_Szilvasi@biogen.com]
Sent:	Wednesday, August 01, 2001 2:25 PM
To:	Cytometry Mailing List
Subject:	The automated cytometer is coming !!!



Just kidding.

But you all opened this mail with excitement. Right?

Why are the cytometers not automated?
Why is the 96 well plate not the default sample carrier?

Most of us stain cell for flow cytometry analysis on 96 well plates. For
over a decade we had to transfer them into tubes, sit in front of an
instrument and exchange them one by one for hours. OK...I am somewhat
emotional today after running close to 600 tubes by hand. My back hurts,
my eyes are tired from the screen and my left shoulder suffered
repetitive motion injury.

Us who run core labs especially in industrial settings have to screen
large volumes of samples on a regular basis. The request for 96 well
sampler was
out there for a very long time. Years. No response from the manufacturers.

Recently BD introduced an addition to the FACS Calibur that can take up
cells from the 96 well format. It is not a great help for those of us
who need to run hundreds a day. Also too expensive and meant for the
hobbyists.

CyTek's MAS is a faster device with limited acquisition control and no
field service.

Cytomation's MosKeeto is only good for simple yes-or-no type screening on a
time
scale on the MoFlo. CyAn does not even have a plate sampler addition (maybe
next year???
- we heard that before).

Why don't we unite and give an ultimatum to the manufacturers. YOU EITHER
MAKE FAST, ROBUST 96 WELL PLATE SAMPLERS OR ELSE .......... we keep
using tubes just like we do now. This is our last, desperate resort.
Flow cytometers are in the forefront of academic and industrial research
as well as manufacturing. The technology (including the software!) is
not following the trend.

I go back and finish the two hundred remaining samples..... (The numbers
are a result of theatrical dramatization - run 800 samples on a Calibur
in a day and your left arm will be paralyzed. You better seek mental help
at the end of such a day.)

Best regards,

Akos
Biogen



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