Fig. 2: Illustration of the workflow for assessing total brain drug concentrations using qMSI-uD. | Molecular Psychiatry

Fig. 2: Illustration of the workflow for assessing total brain drug concentrations using qMSI-uD.

From: Neuropharmacokinetic visualization of regional and subregional unbound antipsychotic drug transport across the blood–brain barrier

Fig. 2

a Cryosectioning of a brain tissue section (left) and a brain slice (right), collected and frozen after performing in vivo and in vitro experiments, respectively. Thaw-mounting of tissue slices on precooled indium tin oxide-coated glass slides and preparation of three technical replicates from consecutive tissue sections on additional glass slides. b Sample preparation for MALDI-qMSI. Spotting of calibration standard solutions of the drug and quality-control (QC) solutions on the top of the tissue. Automatic spraying of a deuterated analog of the drug onto the tissue as an internal standard (IS) followed by application of the MALDI matrix and acquisition of MALDI-MSI data. c Processing of the acquired data involves selection and visualization of ions of interest and normalization of the data against that for the IS. This is followed by determination of regions of interest (ROI) of calibration-standard spots and selected brain regions based on a reference brain atlas [50] generated using the same tissue sections after staining with hematoxylin and eosin. Finally, a calibration curve is created by plotting the drug’s concentration against the average intensity of the ion, and individual average ion intensities within the region of interest are exported and quantitated using the calibration curve.

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