Serial Two-Photon Tomagraphy: an Automated Method for Ex-Vivo Fluorescence 3D Imaging of Entire Organs with Subcellular Resolution

When: Thursday, December 15, 2011 at 12:00 pm
Where: DA 114
Speaker: Dr. Jason Sutin
Organization: MGH
Sponsor: Biophysical Group Meeting

Classical histology discovered physical structures of cells within an organ reflect their function. Modern optical spectrocopy techniques, such as fluorescence proteins, immunofluorescence staining, and extrinsic fluorescence dyes, when combined with light microscopy, have extended this vision into the molecular regime while still retaining the all-important spatial organization of the tissue. Unfortunately, limited penetration of light through tissue restricts high-resolution optical imaging to thin sections unless alternative methods are employed. Furthermore, acquisition speed and data processing requirements scale cubicly with specimen size, presenting formidable technical challenges to imaging entire organs.

Here we describe a new automated method, which we call serial two-photon (STP) tomography, that achieves high-throughput 3D fluorescence imaging of whole rodent-sized organs by integrating two-photon microscopy and tissue sectioning. Our hybrid optical-mechanical approach opens the door to routine systematic whole-organ studies of rodent models of human disease. Practical applications of STP in neuroanatomy will be shown from work between groups at TissueVision.