A biophotonic approach to measure pH in small volumes in vitro: Quantifiable differences in metabolic flux around the cumulus‐oocyte‐complex (COC)
| Autor: | Malcolm S. Purdey, Andrew D. Abell, Kylie R. Dunning, Erik P. Schartner, Jeremy G. Thompson, Hanna J. McLennan, Avishkar Saini, Tanya M. Monro, Georgina M. Sylvia |
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| Přispěvatelé: | McLennan, Hanna J, Saini, Avishkar, Sylvia, Georgina M, Schartner, Erik P, Dunning, Kylie R, Purdey, Malcolm S, Monro, Tanya M, Abell, Andrew D, Thompson, Jeremy G |
| Rok vydání: | 2019 |
| Předmět: |
Technology
DNA damage General Physics and Astronomy 01 natural sciences General Biochemistry Genetics and Molecular Biology 010309 optics Mice chemistry.chemical_compound fluorescence sensing 0103 physical sciences medicine Extracellular Animals General Materials Science Lactic Acid Chemistry 010401 analytical chemistry cumulus oocyte complex lactic acid pH sensing General Engineering General Chemistry Hydrogen-Ion Concentration Oocyte Photobleaching In vitro biophotonic 0104 chemical sciences Staining Lactic acid medicine.anatomical_structure op-tical fibre Oocytes Biophysics biomarker Follicle Stimulating Hormone Phototoxicity micro-environment |
| Zdroj: | Journal of Biophotonics. 13 |
| ISSN: | 1864-0648 1864-063X |
| DOI: | 10.1002/jbio.201960038 |
| Popis: | Unfertilised eggs (oocytes) release chemical biomarkers into the medium surrounding them. This provides an opportunity to monitor cell health and development during assisted reproductive processes if detected in a non-invasive manner. Here we report the measurement of pH using an optical fibre probe, OFP1, in 5 μL drops of culture medium containing single mouse cumulus oocyte complexes (COCs). This allowed for the detection of statistically significant differences in pH between COCs in culture medium with no additives and those incubated with either a chemical (cobalt chloride) or hormonal treatment (follicle stimulating hormone); both of which serve to induce the release of lactic acid into the medium immediately surrounding the COC. Importantly, OFP1 was shown to be cell-safe with no inherent cell toxicity or light-induced phototoxicity indicated by negative DNA damage staining. Pre-measurement photobleaching of the probe reduced fluorescence signal variability, providing improved measurement precision (0.01-0.05 pH units) compared to previous studies. This optical technology presents a promising platform for the measurement of pH and the detection of other extracellular biomarkers to assess cell health during assisted reproduction. Refereed/Peer-reviewed |
| Databáze: | OpenAIRE |
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