Comparative analysis of sucrose-embedding for whole-body zebrafish MSI by IR-MALDESI

Wang, Mary F.; Ritter, Morgan M.; Kullman, Seth W.; Muddiman, David C.

doi:10.1007/s00216-023-04914-1

Cited by 2 publications

(8 citation statements)

References 39 publications

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“…Zebrafish specimens were provided on the same day as they were embedded in CMC/gelatin for MSI analysis. This protocol follows other previous reports. , …”

Section: Methodsmentioning

confidence: 76%

“…This protocol follows other previous reports. 20,34 After the specimens were PFA-fixed and sucrose-embedded, all zebrafish were prepared and sectioned according to a previously optimized protocol. 33 This embedding protocol was utilized in order to successfully obtain whole-body sections of the fish.…”

Section: ■ Introductionmentioning

confidence: 99%

“…No ice matrix was applied for these tissues, as sucrose-embedding was performed and acted as an energy absorbing matrix. 20,21 After spot size characterization, adjacent sections were run in the following order: ( 1 produced square spot shapes of the same width and height; therefore, they will be referred to as 20 μm, 40 μm, and 60 μm from this point forward. Each section was stained and preserved after analysis for downstream microscopy.…”

Section: ■ Introductionmentioning

confidence: 99%

“…For UV-MALDI, the laser is complemented with an organic matrix co-crystallized with the sample, where energy is absorbed at the same or similar wavelength as the laser (e.g., 337 or 355 nm) . In the case of IR-MALDESI, ice and sucrose-embedding , have been utilized as the energy absorbing matrix, as the laser wavelength for this platform is 2970 nm. Because a smaller wavelength is commonly utilized for MALDI systems, they produce smaller spot sizes compared to IR-MALDESI .…”

Section: Introductionmentioning

confidence: 99%

“…Because a smaller wavelength is commonly utilized for MALDI systems, they produce smaller spot sizes compared to IR-MALDESI . For instance, conventional analyses by IR-MALDESI demonstrate spot sizes near 150 μm. ,, However, UV-MALDI platforms have reported spatial resolutions as low as 1.4 μm under ambient conditions, and others have employed a transmission mode strategy to achieve 1 μm, as demonstrated with a cell model . Although such spatial resolution can be achieved by these platforms, UV-MALDI approaches may utilize spatial resolutions anywhere from 10 to 200 μm depending on the application. , …”

Section: Introductionmentioning

confidence: 99%

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Oversampling for Enhanced Spatial Resolution of Zebrafish by Top-Hat IR-MALDESI-MSI

Sohn,

Bowman,

Barnes

et al. 2024

J. Am. Soc. Mass Spectrom.

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Mass spectrometry imaging (MSI) has become a significant tool for measuring chemical species in biological tissues, where much of the impact of these platforms lies in their capability to report the spatial distribution of analytes for correlation to sample morphology. As a result, enhancement of spatial resolution has become a frontier of innovation in the field, and necessary developments are dependent on the ionization source. More particularly, laser-based imaging sources may require modifications to the optical train or alternative sampling techniques. These challenges are heightened for systems with infrared (IR) lasers, as their operating wavelength generates spot sizes that are inherently larger than their ultraviolet counterparts. Recently, the infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) source has shown the utility of a diffractive optical element (DOE) to produce square ablation patterns, termed top-hat IR-MALDESI. If the DOE optic is combined with oversampling methods, smaller ablation volumes can be sampled to render higher spatial resolution imaging experiments. Further, this approach enables reproducible spot sizes and ablation volumes for better comparison between scans. Herein, we investigate the utility of oversampling with top-hat IR-MALDESI to enhance the spatial resolution of measured lipids localized within the head of sectioned zebrafish tissue. Four different spatial resolutions were evaluated for data quality (e.g., mass measurement accuracy, spectral accuracy) and quantity of annotations. Other experimental parameters to consider for high spatial resolution imaging are also discussed. Ultimately, 20 μm spatial resolution was achieved in this work and supports feasibility for use in future IR-MALDESI studies.

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“…Zebrafish specimens were provided on the same day as they were embedded in CMC/gelatin for MSI analysis. This protocol follows other previous reports. , …”

Section: Methodsmentioning

confidence: 76%

Section: ■ Introductionmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%