Fluorescence microscopy imaging of mitochondrial metabolism in cancer cells
Mitochondrial metabolism is an important contributor to cancer cell survival and proliferation that coexists with enhanced glycolytic activity. Measuring mitochondrial activity is useful to characterize cancer metabolism patterns, to identify metabolic vulnerabilities and to identify new drug target...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-06-01
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| Series: | Frontiers in Oncology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fonc.2023.1152553/full |
| _version_ | 1797797721277464576 |
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| author | Monika Gooz Monika Gooz Eduardo N. Maldonado Eduardo N. Maldonado |
| author_facet | Monika Gooz Monika Gooz Eduardo N. Maldonado Eduardo N. Maldonado |
| author_sort | Monika Gooz |
| collection | DOAJ |
| description | Mitochondrial metabolism is an important contributor to cancer cell survival and proliferation that coexists with enhanced glycolytic activity. Measuring mitochondrial activity is useful to characterize cancer metabolism patterns, to identify metabolic vulnerabilities and to identify new drug targets. Optical imaging, especially fluorescent microscopy, is one of the most valuable tools for studying mitochondrial bioenergetics because it provides semiquantitative and quantitative readouts as well as spatiotemporal resolution of mitochondrial metabolism. This review aims to acquaint the reader with microscopy imaging techniques currently used to determine mitochondrial membrane potential (ΔΨm), nicotinamide adenine dinucleotide (NADH), ATP and reactive oxygen species (ROS) that are major readouts of mitochondrial metabolism. We describe features, advantages, and limitations of the most used fluorescence imaging modalities: widefield, confocal and multiphoton microscopy, and fluorescent lifetime imaging (FLIM). We also discus relevant aspects of image processing. We briefly describe the role and production of NADH, NADHP, flavins and various ROS including superoxide and hydrogen peroxide and discuss how these parameters can be analyzed by fluorescent microscopy. We also explain the importance, value, and limitations of label-free autofluorescence imaging of NAD(P)H and FAD. Practical hints for the use of fluorescent probes and newly developed sensors for imaging ΔΨm, ATP and ROS are described. Overall, we provide updated information about the use of microscopy to study cancer metabolism that will be of interest to all investigators regardless of their level of expertise in the field. |
| first_indexed | 2024-03-13T03:52:40Z |
| format | Article |
| id | doaj.art-39063f09d3ad4f37a280b9d88ed4c2b8 |
| institution | Directory Open Access Journal |
| issn | 2234-943X |
| language | English |
| last_indexed | 2024-03-13T03:52:40Z |
| publishDate | 2023-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Oncology |
| spelling | doaj.art-39063f09d3ad4f37a280b9d88ed4c2b82023-06-22T09:20:55ZengFrontiers Media S.A.Frontiers in Oncology2234-943X2023-06-011310.3389/fonc.2023.11525531152553Fluorescence microscopy imaging of mitochondrial metabolism in cancer cellsMonika Gooz0Monika Gooz1Eduardo N. Maldonado2Eduardo N. Maldonado3Department of Drug Discovery & Biomedical Sciences, Medical University of South Carolina, Charleston, SC, United StatesHollings Cancer Center, Medical University of South Carolina, Charleston, SC, United StatesDepartment of Drug Discovery & Biomedical Sciences, Medical University of South Carolina, Charleston, SC, United StatesHollings Cancer Center, Medical University of South Carolina, Charleston, SC, United StatesMitochondrial metabolism is an important contributor to cancer cell survival and proliferation that coexists with enhanced glycolytic activity. Measuring mitochondrial activity is useful to characterize cancer metabolism patterns, to identify metabolic vulnerabilities and to identify new drug targets. Optical imaging, especially fluorescent microscopy, is one of the most valuable tools for studying mitochondrial bioenergetics because it provides semiquantitative and quantitative readouts as well as spatiotemporal resolution of mitochondrial metabolism. This review aims to acquaint the reader with microscopy imaging techniques currently used to determine mitochondrial membrane potential (ΔΨm), nicotinamide adenine dinucleotide (NADH), ATP and reactive oxygen species (ROS) that are major readouts of mitochondrial metabolism. We describe features, advantages, and limitations of the most used fluorescence imaging modalities: widefield, confocal and multiphoton microscopy, and fluorescent lifetime imaging (FLIM). We also discus relevant aspects of image processing. We briefly describe the role and production of NADH, NADHP, flavins and various ROS including superoxide and hydrogen peroxide and discuss how these parameters can be analyzed by fluorescent microscopy. We also explain the importance, value, and limitations of label-free autofluorescence imaging of NAD(P)H and FAD. Practical hints for the use of fluorescent probes and newly developed sensors for imaging ΔΨm, ATP and ROS are described. Overall, we provide updated information about the use of microscopy to study cancer metabolism that will be of interest to all investigators regardless of their level of expertise in the field.https://www.frontiersin.org/articles/10.3389/fonc.2023.1152553/fullfluorescence microscopymitochondrial metabolismNAD(P)HFADmitochondrial membrane potentialROS |
| spellingShingle | Monika Gooz Monika Gooz Eduardo N. Maldonado Eduardo N. Maldonado Fluorescence microscopy imaging of mitochondrial metabolism in cancer cells Frontiers in Oncology fluorescence microscopy mitochondrial metabolism NAD(P)H FAD mitochondrial membrane potential ROS |
| title | Fluorescence microscopy imaging of mitochondrial metabolism in cancer cells |
| title_full | Fluorescence microscopy imaging of mitochondrial metabolism in cancer cells |
| title_fullStr | Fluorescence microscopy imaging of mitochondrial metabolism in cancer cells |
| title_full_unstemmed | Fluorescence microscopy imaging of mitochondrial metabolism in cancer cells |
| title_short | Fluorescence microscopy imaging of mitochondrial metabolism in cancer cells |
| title_sort | fluorescence microscopy imaging of mitochondrial metabolism in cancer cells |
| topic | fluorescence microscopy mitochondrial metabolism NAD(P)H FAD mitochondrial membrane potential ROS |
| url | https://www.frontiersin.org/articles/10.3389/fonc.2023.1152553/full |
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