Summary:
DISCO-MS is a spatial proteomics technology: it enables proteomics analysis of cleared tissues imaged in 3D. DISCO-MS is aided by AI and robotics and yields proteome similar to fresh or fixed samples.
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Step 1:
We start by rendering a mouse body or human organ optically transparent using robust clearing methods such as vDISCO or SHANEL. Then, after their cell-level light-sheet microscopy scan, we can visualize fluorescently-labeled structures in 3D as complete.

Step 2:
Next, we employ deep learning-based image analysis to identify regions associated with illnesses such as Alzheimer’s disease, brain injury, or coronary artery disease in an unbiased manner.

Step 3:
Small tissue regions are isolated with the help of a specialized robotic extraction system called DISCO-bot. Samples can currently be as small as 0.014 mm3. Smaller samples (down to 0.0005 mm3 or 60 cells) can be analyzed using laser capture microdissection.

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.

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.