"High-plex 3D Cyclic immunofluorescence in Cleared FFPE Tissue"
The intricate organization and biology of tissues, both healthy and diseased, present a complex landscape that current data collection and computational methods struggle to fully capture. This work proposes a novel approach to bridge this gap by introducing an advanced method for collecting and analyzing high-plex 3D data from millimeters thick human FFPE tissue. This new technique, grounded in the development of 3D cyclic immunofluorescence imaging using Light Sheet Fluorescence Microscopy (LSFM), aims to revolutionize our understanding of key tissue features such as immune-cell interactions and the organization of nerves. The method leverages LSFM's capability for rapid isotropic 3D imaging of cleared specimens, enhanced by protocols for active tissue staining, immersion-based clearing, and specimen mounting. This approach allows for the imaging of cleared tissue across multiple channels and sessions, providing an unprecedented level of detail and data complexity. I will briefly discuss the preliminary work and the challenges from experimental design, acquisition and management of large data from several microscopes.
#Lightsheet #Multiplex