Demonstrating voxel-by-voxel (v×v) single-point calibration in liver tissue by IR-MALDESI quantitative MSI

The quantification of glutathione (GSH) on a per-voxel basis was achieved by homogenously spraying a known amount of stable isotope labeled glutathione (SIL-GSH) on a microscope slide. Next, liver sections were mounted on top of the coated slides and imaged using IR-MALDESI MSI. Statistical analysis has shown high precision for V×V quantification over a wide concentration range, but the accuracy of this method is currently limited due to the sprayer configuration. Results support the feasibility of V×V quantification as evidence by concentration heatmaps. Additionally, voxel-based quantification provides benefits such as high specificity, straightforward sample preparation, and promising initial statistics analysis which will be advantageous for future qMSI studies. , , # Demonstrating voxel-by-voxel (v×v) single-point calibration in liver tissue by IR-MALDESI quantitative MSI

Dataset DOI: [10.5061/dryad.b5mkkwhrk](10.5061/dryad.b5mkkwhrk)

## Description of the data and file structure

Data collected for this manuscript. All files were converted to .mzML and then to .imzML format.

### Files and variables

#### File: VxV_GSH_MS1_1.raw

**Description:** Replicate 1 of VxV quantification. Image of mouse liver at 7 μm where stable isotope labeled version glutathione (SIL-GSH) was sprayed underneath the liver section and served as a single point calibration at each voxel within the image. The concentration of endogenous GSH (NAT-GSH) was calculated using the Voxel-by-Voxel quantification tool in MSiReader. 

#### File: VxV_GSH_MS1_2.raw

**Description:** Replicate 2 of VxV quantification. The same conditions from the first replicate of VxV quantification apply to replicate 2. 

#### File: SIL_GSH_MS1_1.raw

**Description:** Replicate 1 of spatial curve q...,