[Cytometry] flow cytometry question

Gerstein, Rachel Rachel.Gerstein at umassmed.edu
Mon Feb 3 10:02:08 EST 2014

folks will be able to give you a better answer if the microbiologist could tell you an expected typical size and range of sizes for the bacteria....

Rachel M. Gerstein, Ph.D.
Associate Professor
Department of Microbiology and Physiological Systems
University of Massachusetts Medical School
Albert Sherman Center
Lab ASC8-1005
Office ASC8-1047
WORCESTER MA 01605-2324

From: cytometry-bounces at lists.purdue.edu [cytometry-bounces at lists.purdue.edu] on behalf of Shaw, Pam [pam.shaw at mso.umt.edu]
Sent: Monday, February 03, 2014 9:47 AM
To: Cytometry List
Subject: [Cytometry] FW: flow cytometry question

Hello flow list buddies!

I have a request a bit out of my  comfort zone.  We don’t do a lot of bacterial work, so could use some input on this request (see below).  We have a FACSAria II with 405, 488 and 633 nm lasers.  Would the 633 laser work for cyanobacteria as described below and would we be able to see a difference in size as the request indicates?

Any input would be appreciated!

Thanks in advance.

Pam Shaw
Fluorescence Cytometry Core Facilitator
Center for Environmental Health Sciences
The University of Montana
32 Campus Dr, Skaggs Building 052
Missoula, MT 59812

Ph (406) 243-4974
Email: pamela.shaw at umontana.edu<applewebdata://DE30ADE2-B1DF-44AF-B224-B0B84EDBD782/pamela.shaw@umontana.edu>

From: <Miller>, Scott <Scott.Miller at mso.umt.edu<mailto:Scott.Miller at mso.umt.edu>>
Date: Friday, January 31, 2014 at 12:51 PM
To: "pamela.shaw at umontana.edu<mailto:pamela.shaw at umontana.edu>" <pamela.shaw at umontana.edu<mailto:pamela.shaw at umontana.edu>>
Subject: flow cytometry question

Hi Pam,

By way of introduction, my name is Scott Miller, a microbiologist in DBS, with a research question that may potentially be appropriate for flow cytometry. The gist is that we are comparing wild type and mutant cyanobacteria in one of our projects. Under certain conditions, the mutant appears “clumpier” than the wild type, probably because of differences in extracellular polysaccharides, and we are exploring possible methods for quantifying this. These bacteria autofluoresce if excited with ~590 nm (peak emission at 680 nm), and so we were curious whether it might be feasible to obtain particle size distributions between wild type and mutant by flow cytometry. The bacteria are filamentous and the clumps are very small by macroscopic standards, but I don’t know how this lack of homogeneity of the sample would impact the method. Many thanks for any advice that you might have and for your thoughts on the feasibility of the approach for this question.



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