Application of a ImageJ-Based Method to Measure Blood Flow in Adult Zebrafish and Its Applications for Toxicological and Pharmacological Assessments

Blood flow is an important physiological endpoint to measure cardiovascular performance in animals. Because of their innate transparent bodies, zebrafish is an excellent animal model for assessing in vivo cardiovascular performance. Previously, various helpful methods for measuring blood flow in zeb...

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Main Authors: Ferry Saputra, Tzu-Ming Tseng, Franelyne P. Casuga, Yu-Heng Lai, Chih-Hsin Hung, Chung-Der Hsiao
Format: Article
Language:English
Published: MDPI AG 2025-01-01
Series:Biology
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Online Access:https://www.mdpi.com/2079-7737/14/1/51
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author Ferry Saputra
Tzu-Ming Tseng
Franelyne P. Casuga
Yu-Heng Lai
Chih-Hsin Hung
Chung-Der Hsiao
author_facet Ferry Saputra
Tzu-Ming Tseng
Franelyne P. Casuga
Yu-Heng Lai
Chih-Hsin Hung
Chung-Der Hsiao
author_sort Ferry Saputra
collection DOAJ
description Blood flow is an important physiological endpoint to measure cardiovascular performance in animals. Because of their innate transparent bodies, zebrafish is an excellent animal model for assessing in vivo cardiovascular performance. Previously, various helpful methods for measuring blood flow in zebrafish larvae were discovered and developed. However, an optimized method to measure blood flow in adult zebrafish has not been reported. In this paper, the tail fin region was selected as target for blood flow measurements using the Trackmate method, provided by ImageJ platform. Based on power statistic calculations, the aortic vessel at the tail base was selected, and other parameters, such as ambient temperature, were investigated for method standardization, in order to minimize experimental variation. The method was also validated using fenpropathrin and ponatinib, which showed some cardiac alterations in a previous zebrafish study. We also checked the versatility of this method by following the same setup in black tetra and medaka and found that this method performed well. However, our results show that heavy pigmentation, like that found in tiger barb, and overlapping vessels, like those in parrot fish, make it hard for this method to perform well. Overall, an optimized protocol was used for the first time to measure blood flow velocity in adult wild-type zebrafish without the aid of transgenic lines or fluorescent dye.
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spelling doaj-art-39eb011c94fc49709a0a7a37d025a6cb2025-01-24T13:23:26ZengMDPI AGBiology2079-77372025-01-011415110.3390/biology14010051Application of a ImageJ-Based Method to Measure Blood Flow in Adult Zebrafish and Its Applications for Toxicological and Pharmacological AssessmentsFerry Saputra0Tzu-Ming Tseng1Franelyne P. Casuga2Yu-Heng Lai3Chih-Hsin Hung4Chung-Der Hsiao5Department of Chemistry, Chung Yuan Christian University, Taoyuan 320314, TaiwanSchool of Medicine for International Students, College of Medicine, I-Shou University, Kaohsiung 82445, TaiwanDepartment of Pharmacy, Research Center for the Natural and Applied Science, University of Santo Tomas, Manila 1008, PhilippinesDepartment of Chemistry, Chinese Culture University, Taipei 11114, TaiwanInstitute of Biotechnology and Chemical Engineering, I-Shou University, Kaohsiung 84001, TaiwanDepartment of Chemistry, Chung Yuan Christian University, Taoyuan 320314, TaiwanBlood flow is an important physiological endpoint to measure cardiovascular performance in animals. Because of their innate transparent bodies, zebrafish is an excellent animal model for assessing in vivo cardiovascular performance. Previously, various helpful methods for measuring blood flow in zebrafish larvae were discovered and developed. However, an optimized method to measure blood flow in adult zebrafish has not been reported. In this paper, the tail fin region was selected as target for blood flow measurements using the Trackmate method, provided by ImageJ platform. Based on power statistic calculations, the aortic vessel at the tail base was selected, and other parameters, such as ambient temperature, were investigated for method standardization, in order to minimize experimental variation. The method was also validated using fenpropathrin and ponatinib, which showed some cardiac alterations in a previous zebrafish study. We also checked the versatility of this method by following the same setup in black tetra and medaka and found that this method performed well. However, our results show that heavy pigmentation, like that found in tiger barb, and overlapping vessels, like those in parrot fish, make it hard for this method to perform well. Overall, an optimized protocol was used for the first time to measure blood flow velocity in adult wild-type zebrafish without the aid of transgenic lines or fluorescent dye.https://www.mdpi.com/2079-7737/14/1/51blood flowImageJTrackmatezebrafish
spellingShingle Ferry Saputra
Tzu-Ming Tseng
Franelyne P. Casuga
Yu-Heng Lai
Chih-Hsin Hung
Chung-Der Hsiao
Application of a ImageJ-Based Method to Measure Blood Flow in Adult Zebrafish and Its Applications for Toxicological and Pharmacological Assessments
Biology
blood flow
ImageJ
Trackmate
zebrafish
title Application of a ImageJ-Based Method to Measure Blood Flow in Adult Zebrafish and Its Applications for Toxicological and Pharmacological Assessments
title_full Application of a ImageJ-Based Method to Measure Blood Flow in Adult Zebrafish and Its Applications for Toxicological and Pharmacological Assessments
title_fullStr Application of a ImageJ-Based Method to Measure Blood Flow in Adult Zebrafish and Its Applications for Toxicological and Pharmacological Assessments
title_full_unstemmed Application of a ImageJ-Based Method to Measure Blood Flow in Adult Zebrafish and Its Applications for Toxicological and Pharmacological Assessments
title_short Application of a ImageJ-Based Method to Measure Blood Flow in Adult Zebrafish and Its Applications for Toxicological and Pharmacological Assessments
title_sort application of a imagej based method to measure blood flow in adult zebrafish and its applications for toxicological and pharmacological assessments
topic blood flow
ImageJ
Trackmate
zebrafish
url https://www.mdpi.com/2079-7737/14/1/51
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