Abstract
Zebrafish embryos provide a unique opportunity to visualize complex biological processes, yet conventional imaging modalities are unable to access intricate biomolecular information without compromising the integrity of the embryos. Here, we report the use of confocal Raman spectroscopic imaging for the visualization and multivariate analysis of biomolecular information extracted from unlabeled zebrafish embryos. We outline broad applications of this method in: (i) visualizing the biomolecular distribution of whole embryos in three dimensions, (ii) resolving anatomical features at subcellular spatial resolution, (iii) biomolecular profiling and discrimination of wild type and ΔRD1 mutant Mycobacterium marinum strains in a zebrafish embryo model of tuberculosis and (iv) in vivo temporal monitoring of the wound response in living zebrafish embryos. Overall, this study demonstrates the application of confocal Raman spectroscopic imaging for the comparative bimolecular analysis of fully intact and living zebrafish embryos.
Highlights
Zebrafish embryos provide a unique opportunity to visualize complex biological processes, yet conventional imaging modalities are unable to access intricate biomolecular information without compromising the integrity of the embryos
The work presented here demonstrates how high-resolution confocal Raman spectroscopic imaging can be employed for analysis of whole-mount zebrafish embryos. cRSI offers an unsurpassed combination of z-resolution and signal-to-noise ratio[24], features that are highly advantageous for the biomolecular analysis of zebrafish embryos
We show that volumetric cRSI can distinguish between lesions formed from two different Mycobacterium marinum strains in a zebrafish embryo model of tuberculosis
Summary
Zebrafish embryos provide a unique opportunity to visualize complex biological processes, yet conventional imaging modalities are unable to access intricate biomolecular information without compromising the integrity of the embryos. Raman spectroscopic imaging is a microscopy technique that utilizes inelastic scattering of laser light to generate spectroscopic datasets, which can be used to directly map chemical bonds in unlabeled samples[15] This method can be used to locally identify a broad range of biomolecules using a single excitation wavelength laser, and when combined with multivariate component analysis, it can be used to resolve complex biomolecular features with subcellular spatial resolution[16]. Oshima et al demonstrated the use of light sheet-excited spontaneous Raman spectroscopy (LSDRS) to image live medaka fish (Oryzias latipes) embryos, a related fish model This technique was limited by a low spatial resolution and weak signal[23]. This study shows how Raman-based imaging enables comparative biomolecular analysis using zebrafish embryo models
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
