Cell volume changes as revealed by fluorescence microscopy: Global vs local approaches
| Autor: | Jan Malínský, Helena Pivoňková, Zuzana Heřmanová, Denisa Kirdajova, Thuraya Awadová, Miroslava Anděrová |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Fluorophore Hypertonic Solutions Cell Context (language use) Mice 03 medical and health sciences chemistry.chemical_compound Imaging Three-Dimensional 0302 clinical medicine Extracellular Fluorescence microscope medicine Animals Cells Cultured Cell Size Fluorescent Dyes Cerebral Cortex Microscopy Confocal Osmotic concentration General Neuroscience 3T3 Cells Fibroblasts Calcein 030104 developmental biology medicine.anatomical_structure Hypotonic Solutions Microscopy Fluorescence chemistry Volume (thermodynamics) Astrocytes Biophysics Isotonic Solutions 030217 neurology & neurosurgery |
| Zdroj: | Journal of Neuroscience Methods. 306:38-44 |
| ISSN: | 0165-0270 |
| Popis: | Background Several techniques for cell volume measurement using fluorescence microscopy have been established to date. In this study, we compare the performance of three different approaches which allow for estimations of the cell volume changes in biological samples containing individual fluorescently labeled cells either in culture or in the tissue context. The specific requirements, limitations and advantages of individual approaches are discussed. New Method Global morphometric data are quantitatively compared with local information about the overall cell volume, represented by the concentration of a mobile fluorophore accumulated within the monitored cell. Results Volume changes induced by variations in the extracellular osmolarity in murine fibroblasts and astrocytes either in the culture or in the acute brain slices were registered by the three- and two-dimensional morphometries and by local fluorescence intensity measurements. The performance of the latter approach was verified using FRAP assessment of the fluorophore mobility. Significantly lower amplitudes of the cortical astrocytes swelling were detected by three-dimensional morphometry, when compared to the other two approaches. Consequently, it failed to detect temperature-induced cell volume changes. Comparison with Existing Method(s) The three most popular methods of cell volume measurement are compared to each other in this study. Conclusions We show that the effectivity of global morphometry-based volumetric approaches drops with the increasing cell shape complexity or in the tissue context. In contrast to this, the performance of local fluorescence intensity monitoring, which is also fully capable of reflecting the instant cell volume variations remains stable, independent of the system used and application. |
| Databáze: | OpenAIRE |
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