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------------------------------Original message -----------------------------
>I have some questions concerning cleaning and maintaining a BD FACScan.
>Unfortunately, changing the technicians maintaining and sometimes repairing
>the FACScan obviously implies changes in cleaning and maintenance
>recommendations and regulations. Initially, following procedure for
>cleaning the hydraulic system and measurement capillary on a regular basis
>was recommended:
>
............[edited]...........................................
>
>If any of you have some idea, information, or experiences indicating the
>right and correct (and even the best functioning) method of cleaning the
>FACScan, please drop a note. Any incoming suggestions or comments are
>appreciated.
>
----------------------------------Reply---------------------------------------
A number of threads related to your questions can be found in the archive
section, most recently, see postings "Re: Clean Macines" on 18, 21 and 26
February of this year. Specific cleaning agents and protocols are given.
One area that wasn't mentioned in previous postings or given much attention
in the manual is sample handling an changing of samples. By observing users
of our core facility, we were able to identify certain techniques that
could lead to clogging. The following points are stressed during operator
training and has eliminated clogging:
A.) Avoid using Standby unless the system has been cleaned with Sodium
Hypochlorite (0.5% dilution) for 10 minutes.
B.) Whenever running samples, do sample tube changes quickly, especially if
you are running DNA samples. Give a slight shake of the sample tube and
look for clumping of nuclei or cells, particularly if you are running DNA.
We use a special 6 ml, 12x75mm tube from the Falcon division of B-D that
has a 35 micron filter within the cap (Part Nr. 2235). For bulk filtering
of sample preps, Falcon makes larger diameter filters for centrifuge tubes
(P/N 2340 = 40 micron and P/N 2350 = 70 micron).
C.) If sample changes are slow (longer than 10 seconds), place a tube of
dH20 on the sample uptake capillary to keep fluid running up through the
bore.
D.) After the last sample is acquired, place a tube of dH20 on for 20-30
seconds to flush out any remaining cells and protein debris. This will also
prevent protein becoming gelatinous when it comes in contact with Sodium
Hypochlorite.
E). Always leave a tube of dH2O on the uptake port, particularly on
shut-down to keep the capillary bore from drying out and formation of salt
crystals.
Steven Merlin
University of Bern
FACS Core Facility
Murtenstraase 31
CH-3010 Bern, Switzerland
Lab: ++41-31-632-8876
Office: ++41-31-632-9960
FAX: ++41-31-381-8764
e-mail: merlin@patho.unibe.ch
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
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