A laser diffraction system with improved sensitivity for long-time measurements of sarcomere dynamics in isolated cardiac myocytes

Abstract

To measure the mechanical activity of enzymatically isolated mammalian myocytes the principle of laser light diffraction was used. Since the viability of isolated cardiac myocytes showed a marked dependence on the laser power used, an opto-electronic system with improved light sensitivity and low susceptibility to optical noise was developed. The high sensitivity was achieved by a novel approach in the detection of diffraction patterns, that provides a significant reduction of the amount of laser power required. This improvement rendered possible the application of laser diffraction during extended experiments including pharmacological interventions. The static performance of the system, as assessed by means of calibration gratings, showed a resolution in the order of 5 nm for small changes in sarcomere length in the range from 1.2 microns to 2.0 microns. Examples of measurements on resting and contracting cells are presented, and the limitations of the application of the system to biological specimens are discussed.