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Circ Res. 2013 Sep 13;113(7):863-70. doi: 10.1161/CIRCRESAHA.113.301704. Epub 2013 Jul 30.

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

Author information

1
From the Department of Engineering Science.

Abstract

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.

KEYWORDS:

instrumentation; multiphoton fluorescence microscopy; optical imaging; sarcomeres

PMID:
23899961
DOI:
10.1161/CIRCRESAHA.113.301704
[Indexed for MEDLINE]

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