Conclusions:
DISCO-MS combines the complete 3D-imaging data of whole organs and organisms with their unbiased proteome signatures. It works well in preclinical and clinical samples, thus, can be used almost for any biomedical tissue analysis.

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Limitations:
Currently we cannot reach single cell resolution as such proteomics methods are limited. We also work on improving DISCO-bot for smaller tissue isolation.

Example application 4:
DISCO-MS works equally well on clinical samples. We used DISCO-bot-aided DISCO-MS to study coronary artery disease. After identifying all atherosclerotic plaques in the human heart, we studied proteome composition related to plaque development.

Example application 3:
Next, we studied immune cell heterogeneity in the bone marrow. To this end, we used Lsy-M-EGFP mice (labeling immune cells) and identified regional protein expressions in curvy bones: scapula and cranium.

Example application 2:
Our technology enables us to characterize subtle changes, which would be easily missed using standard methods. To this end, we analyzed the first A-beta plaques appearing in the 5xFAD Alzheimer’s disease mouse model from different brain regions.

Example application 1:
To illustrate the utility of DISCO-MS, we first characterized microglia activation after traumatic brain injury. We identified numerous proteins that were differentially regulated between equivalent regions along the optical tract in damaged and healthy brains.

Step 4:
Next, the molecular composition of the tissue isolates is analyzed using high-sensitivity mass-spectrometry. The cellular proteome measured in cleared tissue is almost indistinguishable from the one obtained from fresh or fixed tissue controls.