Staining of cellular mitochondria with LDS-751

We have found the dye LDS-751 to bind almost exclusively to mitochondria when incubated with viable, nucleated cells. Treatment of cells with the nuclear stain acridine orange and LDS-751 revealed little colocalization when the cells were examined by confocal microscopy. Staining with the dye rhodam...

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Bibliographic Details
Published in:Journal of immunological methods 2001-11, Vol.257 (1), p.35-40
Main Authors: Snyder, D.Scott, Small, Pamela L.C
Format: Article
Language:English
Subjects:
Animals
Arsenicals
Biological and medical sciences
Carbonyl Cyanide m-Chlorophenyl Hydrazone
carbonyl cyanide m-chlorophenylhydrazone
Cell Line
Confocal microscopy
Digitonin
Fluorescent Dyes
Fundamental and applied biological sciences. Psychology
Fundamental immunology
LDS-751
Membrane Potentials
Mice
Microbiology
Microscopy, Confocal
Mitochondria
Mitochondria - ultrastructure
Molecular immunology
Monocytes - ultrastructure
Organic Chemicals
phenyl arsine oxide
Rhodamine 123
Staining
Staining and Labeling - methods
Techniques
Vertebrates: skin, associated glands, phaneres, light organs, various exocrine glands (salt gland, uropygial gland...), adipose tissue, connective tissue
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Summary:We have found the dye LDS-751 to bind almost exclusively to mitochondria when incubated with viable, nucleated cells. Treatment of cells with the nuclear stain acridine orange and LDS-751 revealed little colocalization when the cells were examined by confocal microscopy. Staining with the dye rhodamine 123, which is known to bind polarized mitochondria, was virtually identical to the pattern observed with LDS-751. This staining pattern was observed to be consistent over a range of 0.02–20 μg/ml LDS-751 and was consistent between both fibroblasts and monocytes. Depolarization of mitochondria with the mitochondrial depolarizing agents phenyl arsine oxide and carbonyl cyanide m-chlorophenylhydrazone (CCCP) dramatically reduced both LDS-751 staining, and rhodamine 123 fluorescence. Taken together, these results suggest that LDS-751 is excluded from the nucleus and binds the polarized membranes of mitochondria. Given this, interpretation of LDS-751 fluorescence as being indicative of nuclear status, as is commonly done to discriminate between leukocytes and erythrocytes, is unwarranted and may lead to erroneous conclusions if mitochondria become depolarized upon processing.
ISSN:0022-1759
1872-7905
DOI:10.1016/S0022-1759(01)00440-9