Detection of mitochondrial DNA in living animal cells with fluorescence microscopy.
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| Title: | Detection of mitochondrial DNA in living animal cells with fluorescence microscopy. |
|---|---|
| Authors: | Dellinger, M.1, Gèze, M.1 |
| Source: | Journal of Microscopy. Dec2001, Vol. 204 Issue 3, p196-202. 7p. |
| Subjects: | Mitochondrial DNA, Fluorescence microscopy |
| Abstract: | The detection of mitochondrial DNA (mtDNA) in living human cells could be useful for understanding mitochondrial behaviour during cellular processes and pathological mtDNA depletions. However, until now, human mtDNA has not been visualized in living cells with fluorescence microscopy, although it has been easily detected in organisms with larger mtDNA. Previous reports have stated that mtDNA staining results in homogeneous fluorescence of mitochondria or that animal mitochondria are refractory to DAPI staining. This paper shows that mtDNA of cultured green monkey kidney CV-1 can be stained using a very low concentration of DAPI, then detected by a cooled Photometrics CCD camera with 14-bit resolution detection. Indeed, under these conditions CV-1 cells have small fluorescent spots in the cytoplasm that colocalize with mitochondria, even after mitochondrial movements, uncoupling by carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone and swelling. These observations have been reproduced for the human fibroblast foreskin cell line HS68. These results and known properties of DAPI as a specific DNA stain strongly suggest that mtDNA can be detected and visualized by fluorescence microscopy in human living cells, with potential developments in the study of mtDNA in normal and pathological situations. [ABSTRACT FROM AUTHOR] |
| Copyright of Journal of Microscopy is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.) | |
| Database: | Engineering Source |
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| Items | – Name: Title Label: Title Group: Ti Data: Detection of mitochondrial DNA in living animal cells with fluorescence microscopy. – Name: Author Label: Authors Group: Au Data: <searchLink fieldCode="AR" term="%22Dellinger%2C+M%2E%22">Dellinger, M.</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Gèze%2C+M%2E%22">Gèze, M.</searchLink><relatesTo>1</relatesTo> – Name: TitleSource Label: Source Group: Src Data: <searchLink fieldCode="JN" term="%22Journal+of+Microscopy%22">Journal of Microscopy</searchLink>. Dec2001, Vol. 204 Issue 3, p196-202. 7p. – Name: Subject Label: Subjects Group: Su Data: <searchLink fieldCode="DE" term="%22Mitochondrial+DNA%22">Mitochondrial DNA</searchLink><br /><searchLink fieldCode="DE" term="%22Fluorescence+microscopy%22">Fluorescence microscopy</searchLink> – Name: Abstract Label: Abstract Group: Ab Data: The detection of mitochondrial DNA (mtDNA) in living human cells could be useful for understanding mitochondrial behaviour during cellular processes and pathological mtDNA depletions. However, until now, human mtDNA has not been visualized in living cells with fluorescence microscopy, although it has been easily detected in organisms with larger mtDNA. Previous reports have stated that mtDNA staining results in homogeneous fluorescence of mitochondria or that animal mitochondria are refractory to DAPI staining. This paper shows that mtDNA of cultured green monkey kidney CV-1 can be stained using a very low concentration of DAPI, then detected by a cooled Photometrics CCD camera with 14-bit resolution detection. Indeed, under these conditions CV-1 cells have small fluorescent spots in the cytoplasm that colocalize with mitochondria, even after mitochondrial movements, uncoupling by carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone and swelling. These observations have been reproduced for the human fibroblast foreskin cell line HS68. These results and known properties of DAPI as a specific DNA stain strongly suggest that mtDNA can be detected and visualized by fluorescence microscopy in human living cells, with potential developments in the study of mtDNA in normal and pathological situations. [ABSTRACT FROM AUTHOR] – Name: AbstractSuppliedCopyright Label: Group: Ab Data: <i>Copyright of Journal of Microscopy is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.) |
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| RecordInfo | BibRecord: BibEntity: Identifiers: – Type: doi Value: 10.1046/j.1365-2818.2001.00954.x Languages: – Code: eng Text: English PhysicalDescription: Pagination: PageCount: 7 StartPage: 196 Subjects: – SubjectFull: Mitochondrial DNA Type: general – SubjectFull: Fluorescence microscopy Type: general Titles: – TitleFull: Detection of mitochondrial DNA in living animal cells with fluorescence microscopy. Type: main BibRelationships: HasContributorRelationships: – PersonEntity: Name: NameFull: Dellinger, M. – PersonEntity: Name: NameFull: Gèze, M. IsPartOfRelationships: – BibEntity: Dates: – D: 01 M: 12 Text: Dec2001 Type: published Y: 2001 Identifiers: – Type: issn-print Value: 00222720 Numbering: – Type: volume Value: 204 – Type: issue Value: 3 Titles: – TitleFull: Journal of Microscopy Type: main |
| ResultId | 1 |
