We are interested in experiments where populations of mouse splenocytes are stained with different dyes before mixing and intravenous injection into recipient mice, followed by harvesting the organs and detection of recovered cells by flow. This allows measurement of in-vivo cytotoxicity. For example as described in Hermans et al. JIM 285 25-40 2004. Labeling with CFSE (FL1) is quite straightforward, but we have found this limits the number of discrete populations we can distinguish by labeling with different concentrations of CFSE, so we need to co-stain with a second dye. We have tried PKH-26 (FL2) but this seems to have significant spill-over into the FL1 channel. The dye CMTMR (Molecular Probes C-2927) seems not to be excited by the FACSCalibur lasers (488nm and 635nm) in our tests. From the manufacturer's datasheets the abs wavelength of this dye is 541 and the emission 565. Can anyone recommend a persistent (at least overnight) dye for this application that can be used with the FACSCalibur or FACSCanto and which will give a signal at a longer wavelength than 519nm without spillover into the FL1 channel? Something in the region of APC (660nm) would be ideal. Thanks, Simon F. Lacey Beckman Research Institute of the City of Hope Duarte, CA ----------------------------------------------------------- SECURITY/CONFIDENTIALITY WARNING: This message and any attachments are intended solely for the individual or entity to which they are addressed. This communication may contain information that is privileged, confidential, or exempt from disclosure under applicable law (e.g., personal health information, research data, financial information). Because this e-mail has been sent without encryption, individuals other than the intended recipient may be able to view the information, forward it to others or tamper with the information without the knowledge or consent of the sender. If you are not the intended recipient, or the employee or person responsible for delivering the message to the intended recipient, any dissemination, distribution or copying of the communication is strictly prohibited. If you received the communication in error, please notify the sender immediately by replying to this message and deleting the message and any accompanying files from your system. If, due to the security risks, you do not wish to receive further communications via e-mail, please reply to this message and inform the sender that you do not wish to receive further e-mail from the sender. ===========================================================Received on Wed May 12 16:18:01 2004
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