ALS Brain Imaging mass cytometry

ObjectiveThe aim of this study is to investigate the mechanisms of progressive degeneration and loss of motor neurons, using postmortem samples from patients with sporadic ALS. We analyzed affected regions, including the motor cortex, spinal cord, and cerebellum, which has recently been implicated in ALS pathophysiology. By employing imaging mass cytometry (IMC), using Hyperion (Standard BioTools™, https://www.standardbio.com/), which enables visualization of up to 30 protein markers within a single tissue section, we conducted comprehensive single-cell-level profiling of protein expression patterns related to ALS pathology.Through proteomic analysis, we aim to identify brain region–specific disease-associated glia in the ALS brain and elucidate the relevant target cell types, protein markers, and molecular mechanisms involved in disease progression.

SamplesJapanese volunteers of patients with sporadic ALS aged 50 years or older, as well as age-matched healthy controls (both male and female participants) were participated.For ALS patients, samples were obtained from individuals with an official diagnosis. Age at disease onset, duration of disease progression, medical procedures, medication history, and smoking history was collected in advance.Total cases: 12 (motor cortex, cerebellum, and spinal cord)Males and females: 6 each sex (3 ALS patients, 3 controls)

Measurement MethodFormalin-fixed, paraffin-embedded (FFPE) tissue sections with a thickness of 4 μm were prepared from the following three anatomical regions: the primary motor cortex (precentral gyrus of the frontal lobe), lumbar spinal cord (including the anterior horn), and cerebellum.

A panel of 30 metal-tagged antibodies will be generated for imaging mass cytometry using the Hyperion® Imaging System (Standard BioTools). The antibody panel includes markers for ALS-specific pathological proteins as well as various cell-type markers. After metal conjugation, antibodies were applied to human FFPE tissue sections, and imaging mass cytometry was performed.Data were deposited as .mcd format. MCD SmartViewer can be downloaded from https://www.standardbio.com/mcd-smartviewer-download.

Data infoData information is tabulated in .xlsx file below.ALS_Brain Imaging data table 20250703 - Zenodo.xlsx

R-studio IMC spatial data analysisALS_IMC_analysis - MH.RmdIMC_rawdata_csv_Info.xlsx

Matlab Spatial Distance AnalysisALS_Distance_Analysis_20250607GO_deposit.m