***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Try CF405 from Biotium. Bright and robust Vickie Sent from my iPhone > On Aug 21, 2018, at 7:32 AM, Jeff Carmichael <[log in to unmask]> wrote: > > ***** > To join, leave or search the confocal microscopy listserv, go to: > https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=01%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7Cbcb327d3a8c04f89a37d08d60759c94f%7C22340fa892264871b677d3b3e377af72%7C0&sdata=nSw2ovbXMvqk%2FgJG38PxRboLuMklces9f59Bz4mNH6s%3D&reserved=0 > Post images on https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&data=01%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7Cbcb327d3a8c04f89a37d08d60759c94f%7C22340fa892264871b677d3b3e377af72%7C0&sdata=wAIon1cmiOBqtzyqbG7pC6CvL7Vmq4vAqMncvjfBeb8%3D&reserved=0 and include the link in your posting. > ***** > > This has been addressed more than once in the listserv, but below are some > resources to look into the BrilliantViolet fluorophores which are much more > than an order of magnitude brighter than AF405. You could potentially add > two additional channels in this spectral "dead zone". > > Chroma Technology has no commercial interest in these fluorophores, only in > the filter sets we've recommended for detecting them. > > https://na01.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.jacksonimmuno.com%2Ftechnical%2Fproducts%2Fconjugate-selection%2Fbrilliant-violet&data=01%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7Cbcb327d3a8c04f89a37d08d60759c94f%7C22340fa892264871b677d3b3e377af72%7C0&sdata=LdLt%2BA8lXuru65UJ7hIkPFu8NBLix5ImMzQd9rvTYj4%3D&reserved=0 > https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fbit.ly%2FChromaTechnology2MtEdC9&data=01%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7Cbcb327d3a8c04f89a37d08d60759c94f%7C22340fa892264871b677d3b3e377af72%7C0&sdata=BGgpWCMpZXS6SUtjdRcQGUJltwPCACZH59AkKNMTtTE%3D&reserved=0 > https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fbit.ly%2Fbdbiosciences2MEed69&data=01%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7Cbcb327d3a8c04f89a37d08d60759c94f%7C22340fa892264871b677d3b3e377af72%7C0&sdata=Jy1rfLF694ACftcCJ7vjVe4%2FSYscZhGMtCeXyxm2c0o%3D&reserved=0 > > Good luck and please share your results! > Jeff > > *Jeff Carmichael* > *Product & Technical Marketing Manager* > > *[log in to unmask] <[log in to unmask]> | 802-428-2528* > > * <[log in to unmask]>* > > On Mon, Aug 20, 2018 at 7:06 PM, Cromey, Douglas W - (dcromey) < > [log in to unmask]> wrote: > >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=01%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7Cbcb327d3a8c04f89a37d08d60759c94f%7C22340fa892264871b677d3b3e377af72%7C0&sdata=nSw2ovbXMvqk%2FgJG38PxRboLuMklces9f59Bz4mNH6s%3D&reserved=0 >> Post images on https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&data=01%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7Cbcb327d3a8c04f89a37d08d60759c94f%7C22340fa892264871b677d3b3e377af72%7C0&sdata=wAIon1cmiOBqtzyqbG7pC6CvL7Vmq4vAqMncvjfBeb8%3D&reserved=0 and include the link in your posting. >> ***** >> >> Is there a better or preferred "blue" fluorochrome? Mostly this is a >> widefield fluorescence question (using a DAPI cube), but there is the >> possibility for using a confocal with a 405nm laser line. >> >> I'm working with a group that is trying to work around a frustrating >> autofluorescence issue in liver tissue. What we ended up doing on an >> earlier project was not using any secondaries that fluoresced in the green >> channel (no fitc/AF488/GFP) and instead used the green channel on our >> widefield microscope as a control, ratioing this image so we could subtract >> it out of the other channels to remove some of the broad autofluorescent >> background. This worked nicely. In the published images we showed nuclei >> (blue) and two specific stains using AF568 and AF647, with reduced >> background autofluorescence. >> >> The PI now wants to have labelling with three specific fluorochromes, and >> I am advocating using the green channel as the generic "everything lights >> up" channel for context, so we can hopefully skip the Hoechst 3342 or DAPI. >> Since we won't be using DNA dyes, is there a good blue fluorochrome that >> can be used as a secondary for specific labelling of a marker found on an >> intracellular organelle (other than the nucleus)? >> >> Thanks, >> Doug >> >> ------------------------------------------------------------ >> ------------------------------ >> Douglas W. Cromey, M.S. - Associate Scientific Investigator >> Dept. of Cellular & Molecular Medicine, University of Arizona >> 1501 N. Campbell Ave, Tucson, AZ 85724-5044 USA >> >> office: LSN 463 email: [log in to unmask]<mailto: >> [log in to unmask]> >> voice: 520-626-2824 fax: 520-626-2097 >> >> https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fmicroscopy.arizona.edu%2Flearn%2Fmicroscopy-imaging-resources-www&data=01%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7Cbcb327d3a8c04f89a37d08d60759c94f%7C22340fa892264871b677d3b3e377af72%7C0&sdata=mH0tOoxLFkBkFtgJk6jJD%2BjGmq2Q5%2BsKHuEmwMA%2FsCo%3D&reserved=0 >> Home of: "Microscopy and Imaging Resources on the WWW" >> >> UA Microscopy Alliance - https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fmicroscopy.arizona.edu&data=01%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7Cbcb327d3a8c04f89a37d08d60759c94f%7C22340fa892264871b677d3b3e377af72%7C0&sdata=uWfwVk8sepbnrh7g7yQYp%2FpmDpYDXCt%2FTAmVLSyY%2F64%3D&reserved=0< >> https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fmicroscopy.arizona.edu%2F&data=01%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7Cbcb327d3a8c04f89a37d08d60759c94f%7C22340fa892264871b677d3b3e377af72%7C0&sdata=tO696NIe74G4wfkt48glO7Y9YrK4reBk91jpG3fSCvY%3D&reserved=0> >> > > -- > <https://na01.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.chroma.com%2F&data=01%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7Cbcb327d3a8c04f89a37d08d60759c94f%7C22340fa892264871b677d3b3e377af72%7C0&sdata=5DvluSM%2BvyS8UZttrUybkZlMBGmb3xhxgLoeUQZXIK8%3D&reserved=0>CHROMA TECHNOLOGY CORP® > *an employee owned > company* > 10 Imtec Lane, Bellows Falls, Vermont 05101 USA > 800-824-7662 | > FAX: 802-428-2525 > https://na01.safelinks.protection.outlook.com/?url=www.chroma.com&data=01%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7Cbcb327d3a8c04f89a37d08d60759c94f%7C22340fa892264871b677d3b3e377af72%7C0&sdata=sjhDoWYPBipef1Y2eBSTceqLFI3K4u7ICmqi%2FwGDpB4%3D&reserved=0 <https://na01.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.chroma.com%2F&data=01%7C01%7Cvictoria.frohlich%40STJUDE.ORG%7Cbcb327d3a8c04f89a37d08d60759c94f%7C22340fa892264871b677d3b3e377af72%7C0&sdata=5DvluSM%2BvyS8UZttrUybkZlMBGmb3xhxgLoeUQZXIK8%3D&reserved=0> | > [log in to unmask] <mailto:[log in to unmask]> ________________________________ Email Disclaimer: www.stjude.org/emaildisclaimer Consultation Disclaimer: www.stjude.org/consultationdisclaimer
