On Thursday, 17 April 1997, Andreas Christaras inquired: ------------------------------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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