Work

The optimization of sample preparation on zebrafish larvae in vibrational spectroscopy imaging

Year: 2024

Type: article

Abstract: • Zebrafish larvae are a model organism suitable for FT-IR and Raman imaging. • Optimization of the sample preparation process of Zebrafish larvae for vibrational spectroscopic imaging was performed. • Spectroscopic imaging of zebrafish larvae is providing information in disease models, and toxicology research. • FT-IR and Raman imaging provide information in exploratory and targeted research. The zebrafish (Danio rerio) larvae are widely used in biomedical, pharmaceutical, and ecotoxicological studies. Their transparency and translational potential make them particularly valuable for fluorescence imaging. In addition to fluorescence imaging, microspectroscopy, which combines vibrational spectroscopy: Raman or Fourier transform infrared (FT-IR) with microscopy, allows the collection of spatially resolved, label-free information. According to available literature, it was the first application of FT-IR imaging in zebrafish larvae. This study aims to compare different fixation methods for 10-day post-fertilization (dpf) zebrafish larvae using vibrational spectroscopy imaging. Paraformaldehyde (PFA), glutaraldehyde (GA), low temperature, and embedding in gelatin and agarose were investigated. Amides, lipids, and phosphates distribution were more informative in embedded samples but with challenging handling of the sample due to stiffness at −20 °C. FT-IR and Raman mapping revealed that frozen samples had better-preserved tissue structure than chemical fixation. PFA showed uniform amide distribution, while GA treatment exhibited tissue disruptions and denser protein networks in both. Handling of embedded samples is challenging for an operator, but provides more reliable results in developmental biology or disease modeling, compared to chemical treatment.

Source: Spectrochimica Acta Part A Molecular and Biomolecular Spectroscopy

DOI: 10.1016/j.saa.2024.125288

Authors: Grzegorz Kalisz, Barbara Budzynska, Anna Sroka-Bartnicka

Institution: Medical University of Lublin

Language: English

Topic: Spectroscopy Techniques in Biomedical and Chemical Research

Subfield: Biophysics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: hybrid

Estimated APC paid: $3,470

Funder: Uniwersytet Medyczny w Lublinie