(324d) Rapid Infrared Spectroscopic Imaging of Zebrafish Development Toxicology

Infrared (IR) spectroscopic imaging provides label-free coupled spectral and spatial information that has proved useful in a range of biological systems. The shift from glowbar based FTIR imaging to quantum cascade laser (QCL) based discrete frequency IR (DFIR) imaging has enabled time and data efficient imaging of micro-scale samples. Recent developments in 2-dimensional laser scanning direct detection IR microscopy, DFIR-LSM, allows for rapid high spatial resolution whole-slide imaging. DFIR-LSM high resolution whole-slide spectroscopic imaging was demonstrated by studying embryonic and adult zebrafish. Zebrafish and their embryos are high throughput animal models that are often used for toxicology and drug screening. Standard screening methodologies rely on visual inspection for gross morphological defects, fluorescence microscopy of transgenic fish, or non-spatially resolving techniques such as homogenization followed by mass spectrometry. In this study an IR imaging-based atlas of zebrafish development was generated utilizing DFIR-LSM images of wild-type zebrafish embryos resulting in spectroscopic 3D reconstruction of zebrafish embryos. High throughput all organ screening was achieved through design of a tiered sample array. DFIR-LSM was then used to study the impact of nanoplastics and per- and polyfluoroalkyl substances (PFAS) on embryonic zebrafish development. To further enable future DFIR-LSM zebrafish studies and promote collaboration with biologists, a deep learning model was generated for hematoxylin and eosin (H&E) stainless staining of zebrafish embryos.