Katherine Komon
Research Mentor: Not Available Not Available
Mentor Department: Not Available, Not Available
Author(s): Not Available
Session: Session 2 (10:00 AM – 10:50 AM)
Presentation Type: Poster 5
Abstract
Imaging intact biological organs at cellular resolution remains a major challenge due to
the strong light-scattering properties of biological tissue. Cellular networks span millimeter- to
centimeter-scale structures, yet conventional optical imaging is typically limited to depths of a
few hundred microns in uncleared tissue, preventing visualization of intact organs. Here we
develop optimized optical clearing approaches that enable deep, high-resolution imaging across
intact organs including the brain, kidneys, heart, and liver, etc., and across multiple species
including mouse, monkey, sheep, and human tissue.
Following successful perfusion to preserve tissue integrity, organ samples are embedded
in an improved hydrogel matrix that promotes protein cross-linking, immobilizing proteins
within the gel framework to stabilize structural integrity and tissue architecture. A lipase-driven
delipidation protocol then removes lipids that contribute to light scattering, rendering samples
optically transparent throughout. To address residual optical distortion, a refractive index
matching solution is applied to minimize mismatches between the cleared tissue and the
surrounding medium.
Combining these clearing strategies with advanced fluorescence microscopy and
immunohistochemical labeling enables three-dimensional visualization and high-resolution
computerized reconstruction of complex cellular structures across diverse organ systems. Unlike
conventional SDS-based clearing methods, which disrupt antibody interactions and reduce
reporter signal, our approach preserves native GFP fluorescence and cellular antigen integrity
throughout the clearing process. These methods support large-scale analysis of cellular network
organization at imaging depths and scales previously unattainable, providing a broadly
applicable platform for studying biological architecture across species and organ types.
