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.
Imageprep instrument
Manufactured by Bruker
Sourced in United States
The ImagePrep instrument is a versatile laboratory equipment designed for matrix-assisted laser desorption/ionization (MALDI) sample preparation. It automates the process of sample spotting and matrix application, ensuring consistent and reproducible sample preparation for MALDI mass spectrometry analysis.
Automatically generated - may contain errors
2 protocols using imageprep instrument
1
MALDI Tissue Imaging Protocol
2
Tryptic Digest of Antibody-Functionalized Nanopillars
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The antibody-functionalised nanopillar array chip was subjected to tryptic digest prior to analysis. Sequencing-grade trypsin was made to 50 ng/µL in 25 mM ammonium bicarbonate, and sprayed over the chip using a Bruker Imageprep instrument (Bruker, USA). After trypsin deposition, the chip was incubated in a humid environment at 40 °C for 3 h. Subsequently, the chip was sprayed with a matrix solution, 10 g/L α-cyano-4-hydroxycinnamic acid in 50% acetonitrile with 0.2% trifluoroacetic acid. Next, the chip was analysed with a Bruker Ultraflex III MALDI-TOF mass spectrometer (Bruker, USA) in positive linear mode using Flex Imaging 4.0 (Bruker, USA) with a pixel size of 60 µm. Data were collected from 2 k–30 k m/z, at a laser repetition rate of 200 Hz. Data were normalised using the root mean square approach and visualised using Flex Imaging 4.0 (Bruker, USA) and SCILS LAB 2017a software. For the SCILS LAB analysis, the data were imported using a convolution baseline subtraction, and displayed using root mean squared normalisation.
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