Project Grant R24OD037694

ACQUISITION OF A LEICA ARTOS 3D ULTRAMICROTOME - PROJECT SUMMARY: THE VANDERBILT CELL IMAGING SHARED RESOURCE (CISR) REQUESTS FUNDS TO PURCHASE A LEICA ARTOS ULTRAMICROTOME FOR SAMPLE PREPARATION FOR ELECTRON MICROSCOPY (EM). CISR IS AN INSTITUTIONAL, FEE-FOR- SERVICE, ADVANCED MICROSCOPY RESOURCE. THE CISR PROVIDES RESEARCHERS WITH ACCESS TO STATE-OF-THE-ART IMAGING EQUIPMENT WHICH INCLUDES A MODERN FIB-SEM AND TEM. CISR PROVIDES FULL-SERVICE ELECTRON MICROSCOPY SAMPLE PREPARATION INCLUDING SAMPLE EMBEDDING, THICK AND THIN SECTIONING, STAINING, AND IMAGE ACQUISITION FOR EITHER SEM OR TEM. THESE INSTRUMENTS AND SERVICES ARE AVAILABLE TO SUPPORT ANY INVESTIGATOR WITH AN APPOINTMENT AT VANDERBILT UNIVERSITY OR VANDERBILT UNIVERSITY MEDICAL CENTER. THE CISR SERVES OVER 200 LABS PER YEAR, AND PROCESSES APPROXIMATELY 1,000 SEM AND TEM SAMPLES PER YEAR. MANUAL SERIAL SECTIONING FOR 3-D IMAGE RECONSTRUCTION IS CURRENTLY AN ERROR PRONE, LABOR AND TIME INTENSIVE PROCESS THAT SLOWS DOWN WORK FLOW. THE LEICA ARTOS WILL ALLOW AUTOMATED SERIAL SECTIONING OF CELLS AND TISSUES FOR DOWNSTREAM ARRAY TOMOGRAPHY AND SERIAL TOMOGRAPHY TRANSMISSION ELECTRON MICROSCOPY APPLICATIONS. THE ARTOS CAN AUTOMATICALLY CUT TISSUE SERIAL SECTIONS BETWEEN 50-100 NM WITH AN ADVANCED SET OF FEATURES TO ENABLE COLLECTION OF HUNDREDS OF SECTIONS WITHOUT ERROR. THESE FEATURES INCLUDE AUTOMATED RIBBON SECTIONING WITH HORIZONTAL MOVEMENTS AT A SET NUMBER OF SECTIONS, A SPECIALIZED CUTTING KNIFE THAT CAN HOLD AND CUT SECTIONS FOR AN ENTIRE COVERSLIP, A MECHANISM TO RECOVER ALL THESE SECTIONS WITHOUT INTRODUCING WRINKLES, AND A VIBRATION AND ACOUSTIC ISOLATION PLATFORM TO FACILITATE SECTION REPRODUCIBILITY. THESE SERIAL SECTIONS WILL BE USED FOR TISSUE RECONSTRUCTIONS OF UP TO 100,000 MM3 VOLUME AT NANOMETER RESOLUTION. THIS IS NOT CURRENTLY POSSIBLE WITH THE EQUIPMENT AT VANDERBILT. IF FUNDED THE LEICA ARTOS WOULD BE INCORPORATED INTO THE CISR WORKFLOW RUN BY OUR EXPERT CISR STAFF. DR. EVAN KRYSTOFIAK MANAGES THE CISR ELECTRON MICROSCOPY STAFF INCLUDING MARIA VINOGRADOVA. BOTH EVAN AND MARIA ARE EXPERTS IN ULTRAMICROTOMY AND REGULARLY PREPARE THIN AND THICK SECTIONS ON THE CISR'S EXISTING LEICA ULTRAMICROTOMES, INCLUDING MANUALLY PREPARED, SHORT SERIAL SECTION RIBBONS. THE REQUESTED LEICA ARTOS WOULD BE HOUSED IN EXISTING CISR SPACE IN RR-1207 MCN ALONG WITH CURRENT CISR-EM SAMPLE PREPARATION EQUIPMENT. IN SUMMARY, ACQUISITION OF THE LEICA ARTOS WILL ALLOW FOR UNPRECEDENTED VOLUMETRIC IMAGING CAPABILITIES OF BIOLOGICAL SAMPLES AT SCALES AND SPATIAL RESOLUTIONS NOT CURRENTLY AVAILABLE TO VANDERBILT AND WILL FACILITATE RESEARCH ON A DIVERSE SET OF NIH FUNDED RESEARCH.