Kalbacova 2003 Cytometry: Difference between revisions
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{{Publication | {{Publication | ||
|title= | |title=KalbΓ‘covΓ‘ M, VrbackΓ½ M, Drahota Z, MelkovΓ‘ Z (2003) Comparison of the effect of mitochondrial inhibitors on mitochondrial membrane potential in two different cell lines using flow cytometry and spectrofluorometry. Cytometry 52A:110-6. | ||
|info=[http://www.ncbi.nlm.nih.gov/pubmed/12655654 PMID: 12655654] | |info=[http://www.ncbi.nlm.nih.gov/pubmed/12655654 PMID: 12655654 Open Access] | ||
|authors=Kalbacova M, Vrbacky M, Drahota Z, Melkova Z | |authors=Kalbacova M, Vrbacky M, Drahota Z, Melkova Z | ||
|year=2003 | |year=2003 | ||
| Line 16: | Line 16: | ||
aerobic ATP production. Determination of ΞΞ¨m changes by flow cytometry exhibited greater sensitivity than the ones by spectrofluorometry. | aerobic ATP production. Determination of ΞΞ¨m changes by flow cytometry exhibited greater sensitivity than the ones by spectrofluorometry. | ||
|keywords=Green monkey kidney BSC-40, Human cervical carcinoma HeLa G cell lines | |keywords=Green monkey kidney BSC-40, Human cervical carcinoma HeLa G cell lines | ||
|mipnetlab= | |mipnetlab=CZ Prague Houstek J, CZ Hradec Kralove Cervinkova Z | ||
}} | }} | ||
{{Labeling | {{Labeling | ||
Revision as of 15:36, 24 March 2015
| KalbΓ‘covΓ‘ M, VrbackΓ½ M, Drahota Z, MelkovΓ‘ Z (2003) Comparison of the effect of mitochondrial inhibitors on mitochondrial membrane potential in two different cell lines using flow cytometry and spectrofluorometry. Cytometry 52A:110-6. |
Kalbacova M, Vrbacky M, Drahota Z, Melkova Z (2003) Cytometry
Abstract: Background: Determination of mitochondrial membrane potential ((m) is widely used to characterize cellular metabolism, viability, and apoptosis. Changes of ΞΞ¨m induced by inhibitors of oxidative phosphorylation characterize respective contributions of mitochondria and glycolysis to adenosine triphosphate (ATP) synthesis. Methods: ΞΞ¨m in BSC-40 and HeLa G cell lines was determined by flow cytometry and spectrofluorometry. Its changes induced by specific mitochondrial inhibitors were evaluated using 3,3 ΞΞ¨-dihexyloxacarbocyanine iodide (DiOC6(3)), tetramethylrhodamine ethyl ester, and Mito-Tracker Red. Mitochondrial function was further characterized by oxygen consumption. Results: Inhibition of respiration by antimycin A or uncoupling of mitochondria by FCCP decreased ΞΞ¨m in both cell lines. Inhibition of ATP production by oligomycin or atractyloside induced a moderate decrease of ΞΞ¨m in HeLa G cells and an increase of ΞΞ¨m in BSC-40 cells. Statistically significant differences in ΞΞ¨m between the two cell lines were found with both flow cytometry and spectrofluorometry. Respirometry showed higher basal and FCCP-stimulated respiration in BSC-40 cells. Conclusion: Changes of ΞΞ¨m and oxygen consumption showed that BSC-40 cells are more sensitive than HeLa G cells to inhibitors of mitochondrial function, suggesting that BSC-40 cells are more dependent than HeLa G cells on aerobic ATP production. Determination of ΞΞ¨m changes by flow cytometry exhibited greater sensitivity than the ones by spectrofluorometry. β’ Keywords: Green monkey kidney BSC-40, Human cervical carcinoma HeLa G cell lines
β’ O2k-Network Lab: CZ Prague Houstek J, CZ Hradec Kralove Cervinkova Z
Labels: MiParea: Respiration, Instruments;methods
Preparation: Intact cells
Coupling state: ROUTINE
HRR: Oxygraph-2k
Spectrophotometry; Spectrofluorimetry, monkey
