Sinapinic acid (SA; Bruker Daltonics Inc.) was used as the protein MALDI matrix and prepared as a 10 mg/mL solution in a 50:50 solution of acetonitrile and 0.5% trifluoroacetic acid (TFA). An ImagePrep instrument (Bruker Daltonics Inc.) was used to spray a total of 3 mL of matrix solution on each tissue section. The optimal parameters (dry time, incubation time, and thickness) of the ImagePrep instrument were set to obtain a homogeneous matrix crystal on the tissue sections. After matrix application, the homogeneity of the matrix on the tissue was checked using the imaging function of Chip-1000 (Shimadzu Biotech, Kyoto, Japan). The ITO slide containing the tissue section was mounted on an MTP slide adapter (Bruker Daltonics Inc.), which was directly transferred to the MALDI mass spectrometer.
Mtp slide adapter
Manufactured by Bruker
The MTP slide adapter is a laboratory equipment accessory that allows for the conversion of multi-well microplates (MTPs) into standard microscope slides. This adapter provides a convenient way to prepare samples for analysis using microscopy techniques.
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Lab products found in correlation
2 protocols using mtp slide adapter
1
MALDI Tissue Imaging Protocol
2
MALDI-TOF MS Brain Proteomics
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Brains (n=4) were isolated and immediately snap frozen in isopentane (Sigma, USA) at - Ethanol and then stored in a dessicator under vacuum. In order to be analyzed by MALDI TOF MS, samples were covered with sinapinic acid (10mg/mL in 60% acetonitrile and 0.2% trifluoroacetic acid (TFA)) as MALDI matrix sprayed using
ImagePrep device (Bruker Daltonics). The device parameters were optimized for the brain sections, with a final set-up of 20 cycles combining 2,5s of spray, 5s of incubation and 20s of dry time. Finally the slides were mounted into MTP slide adapter (Bruker Daltonics) and transferred to the MALDI-TOF mass spectrometer. Four biologically independent samples were analyzed by MS. IMS spectra were acquired in linear positive ion mode in an UltrafleXtreme MALDI-TOF/TOF mass spectrometer (Bruker Daltonics). MS data were averaged from 300 consecutive laser shots with a frequency of 2,000 shot/s. The raster spatial resolution was 100 µm.
After the MS analysis, hematoxylin & eosin (H&E) staining (Sigma) was performed in each brain section and they were used as reference for colocalization to select the regions of interest (ROI).
ImagePrep device (Bruker Daltonics). The device parameters were optimized for the brain sections, with a final set-up of 20 cycles combining 2,5s of spray, 5s of incubation and 20s of dry time. Finally the slides were mounted into MTP slide adapter (Bruker Daltonics) and transferred to the MALDI-TOF mass spectrometer. Four biologically independent samples were analyzed by MS. IMS spectra were acquired in linear positive ion mode in an UltrafleXtreme MALDI-TOF/TOF mass spectrometer (Bruker Daltonics). MS data were averaged from 300 consecutive laser shots with a frequency of 2,000 shot/s. The raster spatial resolution was 100 µm.
After the MS analysis, hematoxylin & eosin (H&E) staining (Sigma) was performed in each brain section and they were used as reference for colocalization to select the regions of interest (ROI).
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