研究目的
To investigate the effects of fipronil exposure on phospholipid metabolism in zebrafish using AP-MALDI-MS imaging.
研究成果
AP-MALDI-MSI technique effectively observed changes in metabolites distribution induced by fipronil exposure in zebrafish, offering a new method for toxicological studies and metabolic analysis of pesticides in animals.
研究不足
The study focuses on the immediate effects of fipronil exposure on phospholipid metabolism in zebrafish, without long-term exposure effects or detailed mechanistic insights.
1：Experimental Design and Method Selection
The study utilized atmospheric pressure matrix-assisted laser desorption ionization mass spectrometry (AP-MALDI-MS) for imaging whole zebrafish slices exposed to fipronil to analyze changes in phospholipid metabolism.
2：Sample Selection and Data Sources
Adult AB strain zebrafish were exposed to fipronil aqueous solution (100 μg/L) for 12 hours before being euthanized and frozen for sectioning.
3：List of Experimental Equipment and Materials
SHIMADZU iMScope TRIO for MS imaging, freezing-microtome (Leica CM1950), various MALDI matrices (CHCA, DHB, NEDC, DAN), and solvents (methanol, acetonitrile).
4：Experimental Procedures and Operational Workflow
Zebrafish were sectioned into 10 μm thick slices, mounted onto ITO sheets, and stored at -80℃. Matrix was applied using a homemade device, and sections were analyzed using AP-MALDI-MS with optimized laser power and diameter.
5：Data Analysis Methods
MS data were analyzed using Imaging MS solution Ver.1.30 software, with regions of interest (ROI) manually defined for comparison between exposed and untreated groups.
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SHIMADZU iMScope TRIO
iMScope TRIO
SHIMADZU
Mass spectrometry imaging
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Leica CM1950
CM1950
Leica Biosystems Nussloch GmbH
Freezing-microtome
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α-cyano-4-hydroxycinnamic acid
CHCA
Sigma-Aldrich
MALDI matrix
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2,5-dihydroxybenzoic acid
DHB
Sigma-Aldrich
MALDI matrix
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N-(1-naphthyl) ethylenediamine dihydrochloride
NEDC
Sigma-Aldrich
MALDI matrix
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1,5-diaminonapthalene
DAN
Sigma-Aldrich
MALDI matrix
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