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Zebrafish and Lionheart: A Perfect Match

Zebrafish are essential to Dr. Charles (Chaz) Hong’s laboratory at the University of Maryland School of Medicine. Here, these diminutive powerhouses of developmental biology research are used to characterize the chemical biology of vertebrate development. They are also used to discover novel chemical tools to selectively modulate embryonic development. As a matter of fact, the Hong Lab used zebrafish in their discovery of the world’s first bone morphogenetic protein (BMP) inhibitor, called dorsomorphin. This protein was identified by its ability to make the body of the embryos shorter. Dr. Charles (Chuck) Williams, Assistant Professor of Medicine, who works with the Hong Lab, studies embryonic development in zebrafish using a forward chemical genetic approach. This approach identifies small molecule compounds that interact with proteins that are important in development. Dr. Williams discovers new biological insights based on how the interaction "breaks" the signals that drive zebrafish development.

At Dr. Williams’ original lab, volunteer high school students gathered basic phenotypic information during summer vacation. While this arrangement served to inspire the next generation of researchers, the student availability was limited, and documentation of results widely varied from student to student. As the lab transitioned to a new institution, Dr. Williams wanted to migrate into a high content imaging system. “We wanted to look at more the whole embryo, phenotypes like neural crest migration and more subtle phenotypes and we wanted the information to be as uniform as possible,” Dr. Williams recalls.

Dr. Williams chose the Agilent BioTek Lionheart FX automated microscope as it offers a deep breadth of capabilities to match the varied assays performed in the laboratory. These assays include time course analysis of zebrafish development, scratch and spheroid invasion assays to look at cancer motility, GPCR and calcium flux assays and more. “The Lionheart’s 1.25x magnification objective is perfect for zebrafish imaging because it perfectly captures the full field of a 96-well microplate, yet it provides enough depth of field that the entire embryo is in focus without having to z-stack images,” he explains. Once word spread about the Lionheart FX, other laboratories contacted the Hong Lab for help with assay development and pilot studies using high content imaging. Demand was so high that three months after the Lionheart FX was installed, the lab had to begin an online scheduling plan for those wanting to use the automated microscope. The lab had to anticipate acquiring a second Lionheart FX in the future.

User feedback awards high marks for ease of use. Dr. Williams comments that users of any skill level can easily build out an assay with defined parameters to obtain high-quality data. Even in the case of a new collaborator working with suboptimal fluorophores, Lionheart FX was able to provide the preliminary data needed for the collaborator to obtain a grant. The lab is developing an algorithm that utilizes convolutional neural networks in artificial intelligence to discern between a normal and abnormal embryo, and to also automatically sub-classify the abnormality to completely eliminate observer bias. Dr. Williams mentions that the zebrafish community is a tight group, and their collaborators have all fallen in love with Lionheart FX, adding, “it’s hard to enumerate all the benefits; it’s just a workhorse that fulfills our needs perfectly.”

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Zebrafish

Zebrafish Tg(Fli:EGFP;her4:RFP) image, taken with the Lionheart FX




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To learn more about the Hong Lab at the University of Maryland School of Medicine, visit their website. Thanks to Dr. Charles Williams for sharing his Agilent BioTek experience.

For Research Use Only. Not for use in diagnostic procedures.

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