[Cytometry] FW: flow cytometry question

Ofir Goldberger immunologywiz at gmail.com
Wed Feb 5 01:42:11 EST 2014

Not sure about the ex/em question, but just wanted to add that flow rate could make a huge difference in FSC/SSC as well as fluorescence. My advice is to work through all the flow rates and find what works best for you. 

Good luck! 
--Composed using a teeny tiny keyboard.--Ofir Goldberger, Ph.D. ? Head of Flow Cytometry Unit
Technion, Faculty of Medicine ? 1 Efron St., Haifa 31096, Israel
Phone: 972-4829-5361 ? Cell: 972-544-853521 ? Fax: 972-4829-5363Suggest a time to meet with my online calendar

 From: "Shaw, Pam" <pam.shaw at mso.umt.edu> To: Cytometry List <cytometry at lists.purdue.edu> Subject: [Cytometry] FW: flow cytometry question Date: 03/02/14, 16:47

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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