Insights into metabolic changes during epidermal differentiation as revealed by multiphoton microscopy with fluorescence lifetime imaging
Abstract Rapid developments in the field of organotypic cultures have generated a growing need for effective and non-invasive methods for quality control during tissue development. In this study, we correlate metabolic changes with epidermal differentiation and demonstrate that multiphoton microscop...
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Nature Portfolio
2025-02-01
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| Series: | Scientific Reports |
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| Online Access: | https://doi.org/10.1038/s41598-025-90101-4 |
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| author | Monika Malak Chen Qian Jeemol James Syam Nair Julie Grantham Marica B. Ericson |
| author_facet | Monika Malak Chen Qian Jeemol James Syam Nair Julie Grantham Marica B. Ericson |
| author_sort | Monika Malak |
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| description | Abstract Rapid developments in the field of organotypic cultures have generated a growing need for effective and non-invasive methods for quality control during tissue development. In this study, we correlate metabolic changes with epidermal differentiation and demonstrate that multiphoton microscopy with fluorescence lifetime imaging (MPM-FLIM) can be applied to monitor epidermal differentiation of keratinocytes with respect to proliferative and differentiated states. In a 2D keratinocyte tissue culture model, increased expression of differentiation markers keratin-1 and keratin-10 was induced with calcium supplementation. An accompanying shift from glycolysis to mitochondrial respiration was detected in metabolic flux assays. Analysis of MPM-FLIM images acquired at 750 nm and 900 nm excitation revealed a decreased relative fraction of intracellular NADH and FAD after high calcium treatment, consistent with increased oxidative phosphorylation. Epidermal differentiation could be monitored over a 96 h period. Discrimination analysis based on k-means clustering generated clusters that correlated well with the duration of high Ca2+ treatment, suggesting that MPM-FLIM can provide useful parameters for monitoring keratinocyte differentiation. |
| format | Article |
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| issn | 2045-2322 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Nature Portfolio |
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| series | Scientific Reports |
| spelling | doaj-art-3c422905ca4e4d6794f35761cbb748ef2025-08-20T02:15:00ZengNature PortfolioScientific Reports2045-23222025-02-0115111310.1038/s41598-025-90101-4Insights into metabolic changes during epidermal differentiation as revealed by multiphoton microscopy with fluorescence lifetime imagingMonika Malak0Chen Qian1Jeemol James2Syam Nair3Julie Grantham4Marica B. Ericson5Department of Chemistry and Molecular Biology, Faculty of Science, University of GothenburgDepartment of Chemistry and Molecular Biology, Faculty of Science, University of GothenburgDepartment of Chemistry and Molecular Biology, Faculty of Science, University of GothenburgInstitute of Neuroscience and Physiology, The Sahlgrenska Academy, University of GothenburgDepartment of Chemistry and Molecular Biology, Faculty of Science, University of GothenburgDepartment of Chemistry and Molecular Biology, Faculty of Science, University of GothenburgAbstract Rapid developments in the field of organotypic cultures have generated a growing need for effective and non-invasive methods for quality control during tissue development. In this study, we correlate metabolic changes with epidermal differentiation and demonstrate that multiphoton microscopy with fluorescence lifetime imaging (MPM-FLIM) can be applied to monitor epidermal differentiation of keratinocytes with respect to proliferative and differentiated states. In a 2D keratinocyte tissue culture model, increased expression of differentiation markers keratin-1 and keratin-10 was induced with calcium supplementation. An accompanying shift from glycolysis to mitochondrial respiration was detected in metabolic flux assays. Analysis of MPM-FLIM images acquired at 750 nm and 900 nm excitation revealed a decreased relative fraction of intracellular NADH and FAD after high calcium treatment, consistent with increased oxidative phosphorylation. Epidermal differentiation could be monitored over a 96 h period. Discrimination analysis based on k-means clustering generated clusters that correlated well with the duration of high Ca2+ treatment, suggesting that MPM-FLIM can provide useful parameters for monitoring keratinocyte differentiation.https://doi.org/10.1038/s41598-025-90101-4Multiphoton microscopyFluorescence lifetime imagingEpidermal differentiationMetabolismLabel-free imaging |
| spellingShingle | Monika Malak Chen Qian Jeemol James Syam Nair Julie Grantham Marica B. Ericson Insights into metabolic changes during epidermal differentiation as revealed by multiphoton microscopy with fluorescence lifetime imaging Scientific Reports Multiphoton microscopy Fluorescence lifetime imaging Epidermal differentiation Metabolism Label-free imaging |
| title | Insights into metabolic changes during epidermal differentiation as revealed by multiphoton microscopy with fluorescence lifetime imaging |
| title_full | Insights into metabolic changes during epidermal differentiation as revealed by multiphoton microscopy with fluorescence lifetime imaging |
| title_fullStr | Insights into metabolic changes during epidermal differentiation as revealed by multiphoton microscopy with fluorescence lifetime imaging |
| title_full_unstemmed | Insights into metabolic changes during epidermal differentiation as revealed by multiphoton microscopy with fluorescence lifetime imaging |
| title_short | Insights into metabolic changes during epidermal differentiation as revealed by multiphoton microscopy with fluorescence lifetime imaging |
| title_sort | insights into metabolic changes during epidermal differentiation as revealed by multiphoton microscopy with fluorescence lifetime imaging |
| topic | Multiphoton microscopy Fluorescence lifetime imaging Epidermal differentiation Metabolism Label-free imaging |
| url | https://doi.org/10.1038/s41598-025-90101-4 |
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