Acridine Orange
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http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi all, A quick question: Does the cell permeant dye acridine orange stain mitochondria? I know there is a derivative of the dye that will stain mitochondria, but I need to know if AO itself will stain mitochondria. Any info would be greatly appreciated! Cheers, Kathy Katherine L. Terry PhD Candidate York University Biology Department Toronto ON M3J 1P3 _________________________________________________________________ |
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Hi Kathy,
Acridine orange is not a good marker of mitochondria (you need a nonyl-acridine for this). Instead, acridine orange will stain lysosomes- because it has affinity to lysosomal membrane. Best, Art On Thu, Sep 11, 2008 at 1:07 PM, Katherine Lynn Terry <[hidden email]> wrote: Search the CONFOCAL archive at |
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http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal We used acridine orange in astrocytes last week with the hope that we'd be labelling secretory vesicles. Instead the cells lit up like christmas trees with mitochondria taking up the dye. -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Katherine Lynn Terry Sent: 11 September 2008 18:08 To: [hidden email] Subject: Acridine Orange Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi all, A quick question: Does the cell permeant dye acridine orange stain mitochondria? I know there is a derivative of the dye that will stain mitochondria, but I need to know if AO itself will stain mitochondria. Any info would be greatly appreciated! Cheers, Kathy Katherine L. Terry PhD Candidate York University Biology Department Toronto ON M3J 1P3 _________________________________________________________________ |
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Hi Simon,
Thank you for your reply. How were you able (perhaps this is a naive question) to verify that the organelles stained were mitochondria? Have you published these observations in the past? I am curious because I am working on a manuscript and would like to discuss uptake of AO in live cells, if that is indeed what I am seeing. Thank you for any additional info you can provide, Kathy > Date: Fri, 12 Sep 2008 09:28:34 +0100 > From: [hidden email] > Subject: Re: Acridine Orange > To: [hidden email] > > Search the CONFOCAL archive at > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > > We used acridine orange in astrocytes last week with the hope that we'd > be labelling secretory vesicles. Instead the cells lit up like > christmas trees with mitochondria taking up the dye. > > > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] On > Behalf Of Katherine Lynn Terry > Sent: 11 September 2008 18:08 > To: [hidden email] > Subject: Acridine Orange > > Search the CONFOCAL archive at > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > > Hi all, > > A quick question: Does the cell permeant dye acridine orange stain > mitochondria? I know there is a derivative of the dye that will stain > mitochondria, but I need to know if AO itself will stain mitochondria. > Any info would be greatly appreciated! > > Cheers, > > > Kathy > > > > Katherine L. Terry > PhD Candidate > York University > Biology Department > Toronto ON M3J 1P3 > _________________________________________________________________ Use Windows Live Messenger to send messages to your buddies on their mobile phones Find out more on our PC to Mobile website |
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As I recall, AO accumulates in acidic compartments, the basis for its lighting up endosomes and lysosomes. This suggests that the astrocyte mitochondria had a low pH.
On Sep 16, 2008, at 9:41 AM, Katherine Lynn Terry wrote:
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Acridine Orange is also a cell-permeant nucleic acid binding dye that emits green fluorescence after binding dsDNA and red fluorescence after binding ssDNA or RNA. The dye also accumulates into acidic organelles where I recall the fluorescence is red. However, acridine orange is very unstable and mediates phototoxicity. Years ago we would watch lysosomes explode during illumination at which time the fluorescence changed from red to green. Not much light was required. For this reason I discourage use of acridine orange since so many other more specific markers of lysosomes, mitochondria and DNA are available. These other dyes should be used instead of AO. The only virtue of AO is that it is relatively cheap, but you're getting what you pay for. John Sandra Masur wrote: Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal As I recall, AO accumulates in acidic compartments, the basis for its lighting up endosomes and lysosomes. This suggests that the astrocyte mitochondria had a low pH. -- John J. Lemasters, MD, PhD Professor and South Carolina COEE Endowed Chair Director, Center for Cell Death, Injury and Regeneration Departments of Pharmaceutical Sciences and Biochemistry & Molecular Biology Medical University of South Carolina QF308 Quadrangle Building 280 Calhoun Street, MSC 140 Charleston, SC 29425 Office: 843-792-2153 Lab: 843-792-3530 Fax: 843-792-1617 Email: [hidden email] |
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