Green fluorescing counterstain?
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Stanislav Vitha |
Green fluorescing counterstain?
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Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear confocalists, do you have suggestions for a general counterstain that would fluoresce in green? I am doing 3D imaging of large number of pollen samples, and would like to increase the signal levels so that my acquisition times become more reasonable. The pollen has been processed (cytoplasm removed), so I am trying to stain the shell (exine/intine) and am not interested in the cytoplasm. Rhodamine B staining worked fine, but the emission is in red. I am after shorter wavelength signal for best resolution. Thank you! Stan Vitha |
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Deborah van der List |
Re: Green fluorescing counterstain?
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Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Stan, Try SYBR green II from Molecular Probes. I use it as a nuclear stain on fixed tissue and use it just as I would with DAPI. I use the Argon laser to scan. I don't know how it will work on your pollen though. Deborah -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Stanislav Vitha Sent: Wednesday, January 30, 2008 4:02 PM To: [hidden email] Subject: Green fluorescing counterstain? Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear confocalists, do you have suggestions for a general counterstain that would fluoresce in green? I am doing 3D imaging of large number of pollen samples, and would like to increase the signal levels so that my acquisition times become more reasonable. The pollen has been processed (cytoplasm removed), so I am trying to stain the shell (exine/intine) and am not interested in the cytoplasm. Rhodamine B staining worked fine, but the emission is in red. I am after shorter wavelength signal for best resolution. Thank you! Stan Vitha |
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In reply to this post by Stanislav Vitha
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal You could try safranin. Seems to stain lots of things and excites well at 488 (even better at 514). Fluorescence peaks in the yellow but it's pretty broad. I believe the old Bio-Rad paper test slides were stained with safranin. Guy -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Stanislav Vitha Sent: Thursday, 31 January 2008 11:02 AM To: [hidden email] Subject: Green fluorescing counterstain? Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear confocalists, do you have suggestions for a general counterstain that would fluoresce in green? I am doing 3D imaging of large number of pollen samples, and would like to increase the signal levels so that my acquisition times become more reasonable. The pollen has been processed (cytoplasm removed), so I am trying to stain the shell (exine/intine) and am not interested in the cytoplasm. Rhodamine B staining worked fine, but the emission is in red. I am after shorter wavelength signal for best resolution. Thank you! Stan Vitha No virus found in this incoming message. Checked by AVG Free Edition. Version: 7.5.516 / Virus Database: 269.19.17/1252 - Release Date: 30/01/2008 8:51 PM |
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George McNamara |
Re: Green fluorescing counterstain?
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In reply to this post by Stanislav Vitha
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Reflected light confocal microscopy is much brighter than using a fluorescent dye. At 07:02 PM 1/30/2008, you wrote: >Search the CONFOCAL archive at >http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > >Dear confocalists, >do you have suggestions for a general counterstain that would fluoresce in >green? >I am doing 3D imaging of large number of pollen samples, and would like to >increase the signal levels so that my acquisition times become more >reasonable. > >The pollen has been processed (cytoplasm removed), so I am trying to stain >the shell (exine/intine) and am not interested in the cytoplasm. > >Rhodamine B staining worked fine, but the emission is in red. I am after >shorter wavelength signal for best resolution. > >Thank you! > > >Stan Vitha George McNamara, Ph.D. University of Miami, Miller School of Medicine Image Core Miami, FL 33010 [hidden email] [hidden email] 305-243-8436 office http://home.earthlink.net/~pubspectra/ http://home.earthlink.net/~geomcnamara/ http://www.sylvester.org/research/SR_lab_analytical.asp?ana=desc (see Analytical Imaging Core Facility) |
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Boswell, Carl A - (cboswell) |
Re: Green fluorescing counterstain?
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Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi Stan, I've had the experience of general cell staining with plain old FITC in a buffer. Since the material was stained with unconjugated rhodamine B, you might give unconjugated FITC a try. Carl Carl A. Boswell, Ph.D. Molecular and Cellular Biology University of Arizona 520-954-7053 FAX 520-621-3709 ----- Original Message ----- From: "George McNamara" <[hidden email]> To: <[hidden email]> Sent: Wednesday, January 30, 2008 8:51 PM Subject: Re: Green fluorescing counterstain? > Search the CONFOCAL archive at > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > > Reflected light confocal microscopy is much brighter than using a > fluorescent dye. > > > > > At 07:02 PM 1/30/2008, you wrote: >>Search the CONFOCAL archive at >>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal >> >>Dear confocalists, >>do you have suggestions for a general counterstain that would fluoresce in >>green? >>I am doing 3D imaging of large number of pollen samples, and would like to >>increase the signal levels so that my acquisition times become more >>reasonable. >> >>The pollen has been processed (cytoplasm removed), so I am trying to stain >>the shell (exine/intine) and am not interested in the cytoplasm. >> >>Rhodamine B staining worked fine, but the emission is in red. I am after >>shorter wavelength signal for best resolution. >> >>Thank you! >> >> >>Stan Vitha > > > > > > > George McNamara, Ph.D. > University of Miami, Miller School of Medicine > Image Core > Miami, FL 33010 > [hidden email] > [hidden email] > 305-243-8436 office > http://home.earthlink.net/~pubspectra/ > http://home.earthlink.net/~geomcnamara/ > http://www.sylvester.org/research/SR_lab_analytical.asp?ana=desc (see > Analytical Imaging Core Facility) > |
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Stanislav Vitha |
Re: Green fluorescing counterstain?
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In reply to this post by Stanislav Vitha
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal not if you look at an almost vertical feature, e.g., the side of a sphere (or a pollen grain) Stan On Wed, 30 Jan 2008 22:51:40 -0500, George McNamara <[hidden email]> wrote: >Search the CONFOCAL archive at >http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > >Reflected light confocal microscopy is much brighter than using a >fluorescent dye. > > > > >At 07:02 PM 1/30/2008, you wrote: >>Search the CONFOCAL archive at >>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal >> >>Dear confocalists, >>do you have suggestions for a general counterstain that would fluoresce >>green? >>I am doing 3D imaging of large number of pollen samples, and would like to >>increase the signal levels so that my acquisition times become more >>reasonable. >> >>The pollen has been processed (cytoplasm removed), so I am trying to stain >>the shell (exine/intine) and am not interested in the cytoplasm. >> >>Rhodamine B staining worked fine, but the emission is in red. I am after >>shorter wavelength signal for best resolution. >> >>Thank you! >> >> >>Stan Vitha > > > > > > >George McNamara, Ph.D. >University of Miami, Miller School of Medicine >Image Core >Miami, FL 33010 >[hidden email] >[hidden email] >305-243-8436 office >http://home.earthlink.net/~pubspectra/ >http://home.earthlink.net/~geomcnamara/ >http://www.sylvester.org/research/SR_lab_analytical.asp?ana=desc (see >Analytical Imaging Core Facility) >========================================================================= |
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Judy Trogadis |
Re: Green fluorescing counterstain?
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In reply to this post by Guy Cox
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal This is a late reply but I just saw a brief article in the latest Microscopy Today (Vol 16(1):12-15) where they describe whole cell fluorescent stains, without any specificity, and available with 4 different Ex/Em spectra. Sounds like a counterstain to me. Made by Cellomics, a subsidiary of Thermo Fisher Scientific. I think I will try one of the flavours myself to see how well it works. Judy Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 7Queen 30 Bond St. Toronto, ON M5B 1W8, Canada ph: 416-864-6060 x6337 pager: 416-685-9219 fax: 416-864-6043 [hidden email] >>> Guy Cox <[hidden email]> 01/30/08 7:53 PM >>> Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal You could try safranin. Seems to stain lots of things and excites well at 488 (even better at 514). Fluorescence peaks in the yellow but it's pretty broad. I believe the old Bio-Rad paper test slides were stained with safranin. Guy -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Stanislav Vitha Sent: Thursday, 31 January 2008 11:02 AM To: [hidden email] Subject: Green fluorescing counterstain? Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear confocalists, do you have suggestions for a general counterstain that would fluoresce in green? I am doing 3D imaging of large number of pollen samples, and would like to increase the signal levels so that my acquisition times become more reasonable. The pollen has been processed (cytoplasm removed), so I am trying to stain the shell (exine/intine) and am not interested in the cytoplasm. Rhodamine B staining worked fine, but the emission is in red. I am after shorter wavelength signal for best resolution. Thank you! Stan Vitha No virus found in this incoming message. Checked by AVG Free Edition. Version: 7.5.516 / Virus Database: 269.19.17/1252 - Release Date: 30/01/2008 8:51 PM |
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