Dear Kim Lowe, flowing bacteria is a basic task in many labs and there is no problem, even on live cells, as far as you take care to clean the machine correctly. There are many protocols to clean the cytometer, probably as much as the number of labs or so. For cleaning the machine, particularly with live cells, I am used after the analyses, to run in the cytometer : - 2 tubes (3-4 ml) of bleach (10% in filtered freshwater) - 2 tubes (3-4 ml) of Ethanol (70% in filtered freshwater) - 3 tubes (3-4) of filtered freshwater I keep the same protocol than the one used to analyze the bacteria, and I check the forward and right angle light scatter to detect any bacteria. I keep cleaning until the cytometer is clean. Don't forget to clean also the sampling needle (the outside) and the parts in contact with the bacteria (ethanol 70% performs very well in the major part of cases). I'd like to give you one advice too. Before any analysis, first look at the sample and check if your bacteria are not aggregated. If you see aggregates, filaments, or if the sample looks sticky because of the cell density, try to separate the cells (sonication?) or dilute them (if possible). Your sample must be clear if you want to avoid problems with your flow cytometer (clogging, contamination). In few cases I just refused to run the samples because of its aspect. I hope iot helps. Regards Gerald Gerald Gregori, Ph.D Charge de recherche CNRS Laboratoire de Microbiologie, Geochimie et Ecologie Microbienne (CNRS, UMR 6117) Campus de Luminy, Batiment TPR1, case 901 13288 Marseille, cedex 9 France tel: 33 4 91 82 9114 fax: 33 4 91 82 65 48 Lowe, Kimberley wrote: >Hi all, >I'd like to get your opinions on flowing bacteria. We have a 2 laser Calibur and some >people have approached me to flow (analyze - not sort) bacteria on our machine. What do >people think? Would cleaning be a problem? I would definitely recommend that they fix >the bacteria at the very least. >Thank you. >Kim Lowe >Childrens Hospital >Vascular Biology >Boston, MA > > > > >Received on Fri Oct 8 14:38:00 2004
This archive was generated by hypermail 2.1.8 : Sat Oct 09 2004 - 03:12:07 EST
