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.
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: Michael E Nance; Justin T Whitfield; Yi Zhu; Anne K Gibson; Laurin M Hanft; Kenneth S Campbell; Gerald A Meininger; Kerry S McDonald; Steven S Segal; Timothy L Domeier Journal: Am J Physiol Heart Circ Physiol Date: 2015-07-24 Impact factor: 4.733
Authors: Alexander D Corbett; Rebecca A B Burton; Gil Bub; Patrick S Salter; Simon Tuohy; Martin J Booth; Tony Wilson Journal: Front Physiol Date: 2014-10-08 Impact factor: 4.566
Authors: Pablo Lamata; Ramón Casero; Valentina Carapella; Steve A Niederer; Martin J Bishop; Jürgen E Schneider; Peter Kohl; Vicente Grau Journal: Prog Biophys Mol Biol Date: 2014-08-10 Impact factor: 3.667
Authors: Leighton T Izu; Peter Kohl; Penelope A Boyden; Masahito Miura; Tamas Banyasz; Nipavan Chiamvimonvat; Natalia Trayanova; Donald M Bers; Ye Chen-Izu Journal: J Physiol Date: 2020-02-03 Impact factor: 6.228