Fast measurement of sarcomere length and cell orientation in Langendorff-perfused hearts using remote focusing microscopy.

Journal article

RATIONALE: Sarcomere length (SL) is a key indicator of cardiac mechanical function, but current imaging technologies are limited in their ability to unambiguously measure and characterize SL at the cell level in intact, living tissue. OBJECTIVE: We developed a method for measuring SL and regional cell orientation using remote focusing microscopy, an emerging imaging modality that can capture light from arbitrary oblique planes within a sample. METHODS AND RESULTS: We present a protocol that unambiguously and quickly determines cell orientation from user-selected areas in a field of view by imaging 2 oblique planes that share a common major axis with the cell. We demonstrate the effectiveness of the technique in establishing single-cell SL in Langendorff-perfused hearts loaded with the membrane dye di-4-ANEPPS. CONCLUSIONS: Remote focusing microscopy can measure cell orientation in complex 2-photon data sets without capturing full z stacks. The technique allows rapid assessment of SL in healthy and diseased heart experimental preparations.

Authors: Botcherby, E; Corbett, A; Burton, R; Smith, C; Bollensdorff, C

• Publication status: Published
• Journal: Circulation research
• Volume: 113
• Issue: 7
• Pages: 863-870
• Publication date: 2013-09-01
• DOI: 10.1161/circresaha.113.301704
• EISSN: 1524-4571
• ISSN: 0009-7330
• Language: English
• Keywords: Rats, Wistar; Animals; Rats, Sprague-Dawley; Rats; Myocardial Reperfusion; Female; Microscopy, Fluorescence, Multiphoton; Sarcomeres