Abstract
Fluorescence anisotropy (rG) was determined in monolayers of cultured cells grown on glass coverslips under various conditions with 5μM TMA-DPH in Hank's buffered salt solution at 37°C. rG was inversely related to membrane fluidity and increased with density of cells and age in culture. rG (±SD) was different in confluent human fibroblasts (0.348±0.016), rat astrocytes (0.315 ±0.011) and rat Roc-1 hybridoma cells (oligodendrocytes × C6) (0.304 ±0.018). Environmental factors like temperature, Ca++ and osmolarity as well as X-ray irradiation modified anisotropy in human fibroblasts. rG decreased in absence of extracellular Ca++ (0.301±0,027) and under hypotonic conditions (30 mosmol mannitol reduced rG to 0.298 ±0.015). On the other hand rG was shown to increase at deeper temperatures and under hypertonic conditions (550 mosmol mannitol led to rG of 0.388±0.013).
Conclusions: TMA-DPH is a marker of superficial layers of the membrane. Fluorescence anisotropy measurements in situ allow to study cellular and environmental factors in cultured cells and provide a sensitive method for detecting of possible genetic or acquired alterations of the membrane.
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Toplak, H., Hermettor, A. & Wiesmann, U. 57 IN SITU FLUORESCENCE ANISOTROPY MEASUREMENTS IN CULTURED CELLS BY A COVERSLIP TECHNIQUE USING 1-(4-TRIMETHYL-AMMONIUMPHENYL) −6-PHENYL −1, 3, 5− HEXATRIENE (TMADPH). Pediatr Res 24, 270 (1988). https://doi.org/10.1203/00006450-198808000-00083
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DOI: https://doi.org/10.1203/00006450-198808000-00083