High-throughput imaging of adult fluorescent zebrafish with an LED fluorescence macroscope

Jessica S. Blackburn, Sali Liu, Aubrey R. Raimondi, Myron S. Ignatius, Christopher D. Salthouse, David M. Langenau

Research output: Contribution to journalArticlepeer-review

45 Scopus citations

Abstract

Zebrafish are a useful vertebrate model for the study of development, behavior, disease and cancer. A major advantage of zebrafish is that large numbers of animals can be economically used for experimentation; however, high-throughput methods for imaging live adult zebrafish had not been developed. Here, we describe protocols for building a light-emitting diode (LED) fluorescence macroscope and for using it to simultaneously image up to 30 adult animals that transgenically express a fluorescent protein, are transplanted with fluorescently labeled tumor cells or are tagged with fluorescent elastomers. These protocols show that the LED fluorescence macroscope is capable of distinguishing five fluorescent proteins and can image unanesthetized swimming adult zebrafish in multiple fluorescent channels simultaneously. The macroscope can be built and used for imaging within 1 day, whereas creating fluorescently labeled adult zebrafish requires 1 hour to several months, depending on the method chosen. The LED fluorescence macroscope provides a low-cost, high-throughput method to rapidly screen adult fluorescent zebrafish and it will be useful for imaging transgenic animals, screening for tumor engraftment, and tagging individual fish for long-term analysis.

Original languageEnglish
Pages (from-to)229-241
Number of pages13
JournalNature Protocols
Volume6
Issue number2
DOIs
StatePublished - Feb 2011

Bibliographical note

Funding Information:
acknowleDGMents J.S.B. is supported by National Institutes of Health (NIH) grant 5T32CA09216-26. C.D.S. is supported by the Dev and Linda Gupta Fund. D.M.L. is supported by NIH grants K01 AR055619-01A1 and 3 K01 AR055619-03S1, the Alex’s Lemonade Stand Foundation, the Sarcoma Foundation of America, the Hollis Brownstein Research Grant from the Leukemia Research Foundation and by a grant from the Harvard Stem Cell Institute. We thank Northwest Marine Technology for expert technical advice and for providing visible implant elastomers for our pilot experiments.

ASJC Scopus subject areas

  • General Biochemistry, Genetics and Molecular Biology

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