PUBLICATION
Benchtop X-band electron paramagnetic resonance detection of melanin and Nitroxyl spin probe in zebrafish
- Authors
- Makarova, K., Zawada, K., Wiweger, M.
- ID
- ZDB-PUB-220323-28
- Date
- 2022
- Source
- Free radical biology & medicine 183: 69-74 (Journal)
- Registered Authors
- Wiweger, Malgorzata
- Keywords
- 5-DSA, Capillary, Eumelanin, In vivo measurement of free radicles, Melanin, Membrane fluidity, X-band EPR, Zebrafish
- MeSH Terms
-
- Animals
- Electron Spin Resonance Spectroscopy/methods
- Free Radicals/chemistry
- Melanins*/chemistry
- Nitrogen Oxides
- Zebrafish*
- PubMed
- 35314357 Full text @ Free Radic. Biol. Med.
Citation
Makarova, K., Zawada, K., Wiweger, M. (2022) Benchtop X-band electron paramagnetic resonance detection of melanin and Nitroxyl spin probe in zebrafish. Free radical biology & medicine. 183:69-74.
Abstract
EPR spectroscopy is a technique that provides direct information about free radicals in biological systems. So far, X-band EPR was seldomly used for in vivo studies as the small resonator size and high power used to detect EPR signals were unsuitable for living organisms. Here, we report new solutions which lift some limitations and make X-band EPR suitable for an in vivo detection of free radicals in zebrafish - a small laboratory animal that is often used as a model for various studies related to free radicals. We designed specially-shaped glass and quartz capillaries to ensure the zebrafish's safety during the experiments. The optimal EPR spectrometer parameters, safe for zebrafish embryos and sufficient to obtain EPR spectrum, were 4 scans by 20s, 100G sweep, and 0.8 mW power. Combining the specially-shaped capillary with a multi-harmonic analyzer for the EPR spectrometer allowed increasing the time up to 16 scans by 11s and lowering the power to 0.25 mW. As a proof of principle, we demonstrate the detection of melanin radicals and the 5-DSA spin probe in zebrafish larvae. As fish survive the EPR scans, the possibility of performing multiple measurements of free radicals in living zebrafish offers new tools for studies aiming to understand redox biology and membrane-dependent functions in both health and disease.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping