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Automatic detection of adult cardiomyocyte for high throughput measurements of calcium and contractility

Simultaneous calcium and contractility measurements on isolated adult cardiomyocytes have been the gold standard for the last decades to study cardiac (patho)physiology. However, the throughput of this system is low which limits the number of compounds that can be tested per animal. We developed ins...

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Autores principales: Cao, L., Manders, E., Helmes, M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8409674/
https://www.ncbi.nlm.nih.gov/pubmed/34469476
http://dx.doi.org/10.1371/journal.pone.0256713
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author Cao, L.
Manders, E.
Helmes, M.
author_facet Cao, L.
Manders, E.
Helmes, M.
author_sort Cao, L.
collection PubMed
description Simultaneous calcium and contractility measurements on isolated adult cardiomyocytes have been the gold standard for the last decades to study cardiac (patho)physiology. However, the throughput of this system is low which limits the number of compounds that can be tested per animal. We developed instrumentation and software that can automatically find adult cardiomyocytes. Cells are detected based on the cell boundary using a Sobel-filter to find the edge information in the field of view. Separately, we detected motion by calculating the variance of intensity for each pixel in the frame through time. Additionally, it detects the best region for calcium and contractility measurements. A sensitivity of 0.66 ± 0.08 and a precision of 0.82 ± 0.03 was reached using our cell finding algorithm. The percentage of cells that were found and had good contractility measurements was 90 ± 10%. In addition, the average time between 2 cardiomyocyte calcium and contractility measurements decreased from 93.5 ± 80.2 to 15.6 ± 8.0 seconds using our software and microscope. This drastically increases throughput and provides a higher statistical reliability when performing adult cardiomyocyte functional experiments.
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spelling pubmed-84096742021-09-02 Automatic detection of adult cardiomyocyte for high throughput measurements of calcium and contractility Cao, L. Manders, E. Helmes, M. PLoS One Research Article Simultaneous calcium and contractility measurements on isolated adult cardiomyocytes have been the gold standard for the last decades to study cardiac (patho)physiology. However, the throughput of this system is low which limits the number of compounds that can be tested per animal. We developed instrumentation and software that can automatically find adult cardiomyocytes. Cells are detected based on the cell boundary using a Sobel-filter to find the edge information in the field of view. Separately, we detected motion by calculating the variance of intensity for each pixel in the frame through time. Additionally, it detects the best region for calcium and contractility measurements. A sensitivity of 0.66 ± 0.08 and a precision of 0.82 ± 0.03 was reached using our cell finding algorithm. The percentage of cells that were found and had good contractility measurements was 90 ± 10%. In addition, the average time between 2 cardiomyocyte calcium and contractility measurements decreased from 93.5 ± 80.2 to 15.6 ± 8.0 seconds using our software and microscope. This drastically increases throughput and provides a higher statistical reliability when performing adult cardiomyocyte functional experiments. Public Library of Science 2021-09-01 /pmc/articles/PMC8409674/ /pubmed/34469476 http://dx.doi.org/10.1371/journal.pone.0256713 Text en © 2021 Cao et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Cao, L.
Manders, E.
Helmes, M.
Automatic detection of adult cardiomyocyte for high throughput measurements of calcium and contractility
title Automatic detection of adult cardiomyocyte for high throughput measurements of calcium and contractility
title_full Automatic detection of adult cardiomyocyte for high throughput measurements of calcium and contractility
title_fullStr Automatic detection of adult cardiomyocyte for high throughput measurements of calcium and contractility
title_full_unstemmed Automatic detection of adult cardiomyocyte for high throughput measurements of calcium and contractility
title_short Automatic detection of adult cardiomyocyte for high throughput measurements of calcium and contractility
title_sort automatic detection of adult cardiomyocyte for high throughput measurements of calcium and contractility
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8409674/
https://www.ncbi.nlm.nih.gov/pubmed/34469476
http://dx.doi.org/10.1371/journal.pone.0256713
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