RE: [Histonet] Mounting Medium for Immunofluorescence
Actually if I could make a minor correction to your statement: propidium iodide is excited by green light at 488nm but emits in the red portion of the spectrum (620nm)... --On Thursday, May 20, 2010 12:48 PM -0600 Liz Chlipala wrote: I think that the propidium iodide will stain the nuclei or DNA, similar to DAPI, so this mounting media will counterstain the nuclei and its visible in the FITC wavelength (488), while if you conterstain with DAPI it is excited with a different wavelength, therefore it might interfere with the FITC portion of your stain. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Laurie Colbert Sent: Thursday, May 20, 2010 12:13 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mounting Medium for Immunofluorescence We normally coverslip our immunofluorescence slides with a permanent aqueous mounting medium. By mistake, today they were coverslipped with a mounting medium that we use for coverslipping our FISH slides. This mounting medium is described as "Mounting medium for fluorescence, with propidium iodide." Will this make a difference when reading the slides?? Laurie Colbert ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician III Cardiovascular Medicine 348 Biomedical Research Building State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 USA lei...@buffalo.edu 716-829-6118 (Ph) 716-829-2665 (Fx) No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Mounting Medium for Immunofluorescence
Hi Laurie, Propidium iodide is a DNA intercalator that fluoresces red (ex 488nm). Regards, Merced --On Thursday, May 20, 2010 11:13 AM -0700 Laurie Colbert wrote: We normally coverslip our immunofluorescence slides with a permanent aqueous mounting medium. By mistake, today they were coverslipped with a mounting medium that we use for coverslipping our FISH slides. This mounting medium is described as "Mounting medium for fluorescence, with propidium iodide." Will this make a difference when reading the slides?? Laurie Colbert ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician III Cardiovascular Medicine 348 Biomedical Research Building State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 USA lei...@buffalo.edu 716-829-6118 (Ph) 716-829-2665 (Fx) No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Mounting Medium for Immunofluorescence
I think that the propidium iodide will stain the nuclei or DNA, similar to DAPI, so this mounting media will counterstain the nuclei and its visible in the FITC wavelength (488), while if you conterstain with DAPI it is excited with a different wavelength, therefore it might interfere with the FITC portion of your stain. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Laurie Colbert Sent: Thursday, May 20, 2010 12:13 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mounting Medium for Immunofluorescence We normally coverslip our immunofluorescence slides with a permanent aqueous mounting medium. By mistake, today they were coverslipped with a mounting medium that we use for coverslipping our FISH slides. This mounting medium is described as "Mounting medium for fluorescence, with propidium iodide." Will this make a difference when reading the slides?? Laurie Colbert ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Mounting Medium for Immunofluorescence
We normally coverslip our immunofluorescence slides with a permanent aqueous mounting medium. By mistake, today they were coverslipped with a mounting medium that we use for coverslipping our FISH slides. This mounting medium is described as "Mounting medium for fluorescence, with propidium iodide." Will this make a difference when reading the slides?? Laurie Colbert ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet