Fast measurement of sarcomere length and cell orientation in Langendorff-perfused hearts using remote focusing microscopy.
Circ Res
; 113(7): 863-70, 2013 Sep 13.
Article
em En
| MEDLINE
| ID: mdl-23899961
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 ANDRESULTS:
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.Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Sarcômeros
/
Reperfusão Miocárdica
/
Microscopia de Fluorescência por Excitação Multifotônica
Limite:
Animals
Idioma:
En
Ano de publicação:
2013
Tipo de documento:
Article