Fast measurement of sarcomere length and cell orientation in Langendorff-perfused hearts using remote focusing microscopy.
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 ce...
| Main Authors: | , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
2013
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| _version_ | 1826277794246557696 |
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| author | Botcherby, E Corbett, A Burton, R Smith, C Bollensdorff, C Booth, M Kohl, P Wilson, T Bub, G |
| author_facet | Botcherby, E Corbett, A Burton, R Smith, C Bollensdorff, C Booth, M Kohl, P Wilson, T Bub, G |
| author_sort | Botcherby, E |
| collection | OXFORD |
| description | 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. |
| first_indexed | 2024-03-06T23:34:16Z |
| format | Journal article |
| id | oxford-uuid:6d1d9cff-d5fa-49de-9065-3a475875c926 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T23:34:16Z |
| publishDate | 2013 |
| record_format | dspace |
| spelling | oxford-uuid:6d1d9cff-d5fa-49de-9065-3a475875c9262022-03-26T19:15:40ZFast measurement of sarcomere length and cell orientation in Langendorff-perfused hearts using remote focusing microscopy.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:6d1d9cff-d5fa-49de-9065-3a475875c926EnglishSymplectic Elements at Oxford2013Botcherby, ECorbett, ABurton, RSmith, CBollensdorff, CBooth, MKohl, PWilson, TBub, GRATIONALE: 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. |
| spellingShingle | Botcherby, E Corbett, A Burton, R Smith, C Bollensdorff, C Booth, M Kohl, P Wilson, T Bub, G Fast measurement of sarcomere length and cell orientation in Langendorff-perfused hearts using remote focusing microscopy. |
| title | Fast measurement of sarcomere length and cell orientation in Langendorff-perfused hearts using remote focusing microscopy. |
| title_full | Fast measurement of sarcomere length and cell orientation in Langendorff-perfused hearts using remote focusing microscopy. |
| title_fullStr | Fast measurement of sarcomere length and cell orientation in Langendorff-perfused hearts using remote focusing microscopy. |
| title_full_unstemmed | Fast measurement of sarcomere length and cell orientation in Langendorff-perfused hearts using remote focusing microscopy. |
| title_short | Fast measurement of sarcomere length and cell orientation in Langendorff-perfused hearts using remote focusing microscopy. |
| title_sort | fast measurement of sarcomere length and cell orientation in langendorff perfused hearts using remote focusing microscopy |
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