Claude, To excite CFP I use the 457 line from an I-70 on our TSO Vantage. The 453 line actually does a better job to excite CFP but the power is lower. The signal to noise ratio was exactly the same reguardless of which excitation line I used. We use YFP with CFP for dual fluorescent protein experiments. To excite the YFP I have been using the 514 line or our argon ion laser. In my hands the YFP excites very well and there is just a slight amount of cross over of CFP bleeding into my YFP channel but only for CFP signals above 10E3. I have never seen YFP bleed into my GFP detector. I can avoid cross beam compensation problems with my above setup. My emmission filter for CFP is a 482/22. My emmission filter for YFP is a 575/26 (not ideal but it works well enough) Glenn MIT >Content-Type: text/plain; charset=us-ascii; x-mac-type="54455854"; >x-mac-creator="4D4F5353" >Content-Transfer-Encoding: 7bit > >As a newbie to CFP, I would like to ask those who have had the >experience whether I should(not) expect to detect CFP using the UV line >of an Enterprise II laser. I realize it is not ideal... perhaps far >from ideal, but I was wondering if I could get enough excitation. While >I'm at it, is there other experience out there for GHF / CFP together? >I've read Beavis and Kalejta in Cytometry 37, 68-73, so mefears I'm out >of luck with an Enterprise II and Melles Griot HeNe (on a MoFlo). > >Finally, can I get enough energy in the 433 - 453 range from an Innova >70 Spectrum to do the job? If so, can this be done on a FACStar Plus >without cross beam compensation? > >Thanks, > >Claude > >Content-Type: text/x-vcard; charset=us-ascii; > name="cantinc.vcf" >Content-Transfer-Encoding: 7bit >Content-Description: Card for Claude Cantin >Content-Disposition: attachment; > filename="cantinc.vcf" > >Attachment converted: Macintosh HD:cantinc.vcf (TEXT/ttxt) (0002D378)
This archive was generated by hypermail 2b29 : Sat Mar 10 2001 - 19:31:41 EST