Enhancing efficiency of microscopy with zebrafish

Application Notes

Enhancing efficiency of microscopy with zebrafish

Jason Otterstrom¹Ephraim Schwartz¹, Yael Geva¹, Shlomo Turgeman¹, Yael Paran¹

1IDEA Bio-Medical, Rehovot, Israel

How can microscopic analysis of zebrafish embryos and larvae be made easier and more efficient? There are many steps and bottlenecks in this experimental workflow [1], and we focus on two key areas: image analysis and automated image acquisition.  Microscopy permits direct visualization of the biology of interest.  Zebrafish anatomy is often clearly visible to human observers but not readily detected by computers given the contrast of brightfield images.  To increase throughput, robustness and permit unbiased image analysis of zebrafish, we have developed a novel AI-based algorithm to detect the fish contour and internal anatomy in brightfield images.  By identifying these hard to detect structures, our software maintains the anatomical context of associated fluorescence signals to enable true high-content imaging in zebrafish.  Thusly, zebrafish morphology along with region-specific spot/cell counting and fluorescence intensity measurement are readily quantified.  Consistent image acquisition is equally crucian for obtaining reproducible results, which we achieve with our Hermes high-content imaging platform.  It holds a key advantage for imaging zebrafish as it was the first microscope in the market to offer a mobile objective, leaving the sample stationary during imaging.  This gentle sample scanning approach does not disturb zebrafish orientation while the microscope obtains multi-color images, including in Z-stack.  We now look towards other bottlenecks in the experimental workflow, including sample preparation.  However, myriad experimental assays utilize zebrafish samples, and new assays continually being developed across diverse fields.  Hence, more information is needed to identify the way forward.  What are the commonalities among the experimental assays?  Can there be universally applied solutions and how do we get there?  Come share your thoughts and we will discuss ideas.

[1] Otterstrom, J.J., Lubin, A., Payne, E.M., Paran, Y. “Technologies bringing young Zebrafish from a niche field to the limelight.” SLAS Tech (2022) 27, pp. 109-120; DOI: 10.1016/j.slast.2021.12.005.