Optical measurements of intracellular pH and magnesium in frog skeletal muscle fibres

The Journal of Physiology
S M BaylorM W Marshall

Abstract

1. Single twitch fibres were isolated from frog muscle, then mounted in a chamber which was positioned on an optical bench. The fibres were immobilized by high stretch (sarcomere spacing 3.9-4.3 mum) and by placement on a pedestal. Their optical properties were determined by illuminating a 35-65 mum diameter spot with quasimonochromatic light of intensity I(0) and measuring the intensity I of the transmitted light. Since the main purpose of the experiments was to draw inferences from the absorbance spectra of different indicator dyes injected into the fibres, all results were expressed in terms of absorbance A calculated from the equation [Formula: see text]. Changes in absorbance DeltaA were calculated from the differential form of the equation [Formula: see text].2. The absorbance of a normal, non-injected fibre was, on average, equal to 0.03 at 570 nm and varied approximately inversely with wavelength between 450 and 750 nm.3. The earliest change in absorbance following an action potential was a small, transient increase which was followed by a larger decrease. The decrease in fibre absorbance varied from 0.5 x 10(-4) to 3 x 10(-4) units.4. Resting myoplasmic pH was determined by comparing the absorbance spectrum from fibres...Continue Reading

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