We have used two instruments to assess SP cells in bone marrow and peripheral blood. The first is a FACStar Plus with the following laser combination. ILT 100 mw 488 Coherent 300 Series water cooled for UV. 50-100 mw is more than sufficient. Recently we changed the ILT for a Coherent Innova 90. Second is a FACS Vantage SE Enterprise laser regulating in UV at 50mw 488 from this as well. Equivalent date can be obtained from either configuration. one key is good filters. we have purchased from different sources and each have their own characteristics. Some are clearly better in our hands than others. The other is staining. The protocol seems simple in the paper, but it is not trivial. Cell number per unit volume, dye concentration and time are variable factors. Dye concentrations and time vary between species and tissues. Charles A. Kuszynski, Ph.D. Assistant Professor/Director Cell Analysis Facility University of Nebraska Medical Center 986495 Nebraska Medical Center Omaha, NE 68198-6495 402 559-6267 voice 402 559-4077 fax ckuszyns@unmc.edu |---------+----------------------------> | | "Beth Whalen" | | | <BWhalen@mrl.ubc.| | | ca> | | | | | | 11/18/2002 11:40 | | | AM | | | | |---------+----------------------------> >---------------------------------------------------------------------------------------------------------------| | | | To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu> | | cc: | | Subject: UV laser for sorting SP cells | >---------------------------------------------------------------------------------------------------------------| Hi, Our lab is going to start using Margaret Goodells method to look at SP cells. For those of you already doing this can you tell me what type of laser you use for UV? I am looking at using either an Enterprise laser with 488nm and UV or having separate UV and 488nm lasers. Thank you, Beth Whalen Manager, Flow Cytometry Core Facility McDonald Research Laboratories/The iCAPTURE Centre St. Paul's Hospital - University of British Columbia 1081 Burrard Street, Vancouver, BC V6Z 1Y6 P: 604-806-8328 F: 604-806-8351
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