***** To join or leave the confocal microscopy listserv or to change your email address, go to: https://lists.umn.edu/cgi-bin/wa?SUBED1=confocalmicroscopy&A=1 Post images on http://www.imgur.com and include the link in your posting. ***** Immature mCherry could faintly show up in the GFP channel. https://twitter.com/joachimgoedhart/status/1395381367760896001?s=20&t=1HvYjjisjINEd4d7Rd4ZuA [https://pbs.twimg.com/media/E11j8_iWUAI5YdI.png:large]<https://twitter.com/joachimgoedhart/status/1395381367760896001?s=20&t=1HvYjjisjINEd4d7Rd4ZuA> Joáchim Goedhárt on Twitter<https://twitter.com/joachimgoedhart/status/1395381367760896001?s=20&t=1HvYjjisjINEd4d7Rd4ZuA> “@JWietek @EvYemini @irinakitop Yep - here's our data (pre-mScarlet age) of the GFP-component of RFPs (only RFP was expressed in these cells). mCherry shows the least green component of these” twitter.com ________________________________ From: Confocal Microscopy List <[log in to unmask]> on behalf of Cammer, Michael <[log in to unmask]> Sent: Thursday, September 8, 2022 5:12 PM To: [log in to unmask] <[log in to unmask]> Subject: mCherry in blue and green channels? Attention: This email originated outside of Penn State Health. Use caution when clicking links or opening attachments. ***** To join or leave the confocal microscopy listserv or to change your email address, go to: https://urldefense.com/v3/__https://lists.umn.edu/cgi-bin/wa?SUBED1=confocalmicroscopy&A=1__;!!Ls64Rlj6!3ixurMjtJ_JDI_iSGir5as1nZdZE1WrrD4UW8KWu_ITKYmmR67DGaOd7jsDqhEDEKk3dFIv39i3zdxJVgMCXpBpUfUo4I5l3fsbbUOc$ Post images on https://urldefense.com/v3/__http://www.imgur.com__;!!Ls64Rlj6!3ixurMjtJ_JDI_iSGir5as1nZdZE1WrrD4UW8KWu_ITKYmmR67DGaOd7jsDqhEDEKk3dFIv39i3zdxJVgMCXpBpUfUo4I5l3faMnpPo$ and include the link in your posting. ***** We have a question about mCherry emitting in the blue channel. We thought we had a nifty result when imaging with a blue/green probe and using mCherry as an indicator of which samples were positive or negative. But then we looked at mCherry alone controls and saw signal in the blue and green channels. Specifically, when exciting at 405 nm, we see strong signal with a 455/50 filter with a spinning disk or similar spectral range with a Zeiss 700. FPbase does show mCherry absorbs in this range https://urldefense.com/v3/__https://www.fpbase.org/protein/mcherry/__;!!Ls64Rlj6!3ixurMjtJ_JDI_iSGir5as1nZdZE1WrrD4UW8KWu_ITKYmmR67DGaOd7jsDqhEDEKk3dFIv39i3zdxJVgMCXpBpUfUo4I5l3hi2E9Fs$ , but does not show emission below 550 nm. We also see a little signal in the green channel, ex 488 and emission 525/50 (and similar with Zeiss 700). Have other people seen this? Thank you! Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York, NY 10016 [log in to unmask]<mailto:[log in to unmask]> https://urldefense.com/v3/__http://nyulmc.org/micros__;!!Ls64Rlj6!3ixurMjtJ_JDI_iSGir5as1nZdZE1WrrD4UW8KWu_ITKYmmR67DGaOd7jsDqhEDEKk3dFIv39i3zdxJVgMCXpBpUfUo4I5l3f_u_amc$ https://urldefense.com/v3/__http://microscopynotes.com/__;!!Ls64Rlj6!3ixurMjtJ_JDI_iSGir5as1nZdZE1WrrD4UW8KWu_ITKYmmR67DGaOd7jsDqhEDEKk3dFIv39i3zdxJVgMCXpBpUfUo4I5l3GjVuz7k$ Voice direct only, no text or messages: 1-914-309-3270 and 1-646-501-0567
