ACCESS AND AVAILABILITY TO SAFE, SECURE GENETIC RESOURCE COLLECTIONS UNDERPINS AMERICAN AGRICULTURAL. MANY MATERIALS IN OUR NATIONAL PLANT COLLECTIONS ARE VULNERABLE TO ABIOTIC, BIOTIC, AND ENVIRONMENTAL THREATS AND MUST BE BACKED-UP IN A COST-EFFECTIVE MANNER AT SECONDARY LOCATIONS. CRYOPRESERVATION TECHNIQUES HAVE BEEN DEVELOPED AND IMPLEMENTED TO SECURE OUR NATIONAL PLANT COLLECTIONS. THEY ENLIST CRYOPROTECTANT SOLUTIONS THAT REMOVE AND REPLACE WATER, ALLOWING PLANT CELLS TO SURVIVE CRYOGENIC CONDITIONS. THESE SAME TREATMENTS MAY SELECTIVELY ELIMINATE VIRUSES AND BACTERIA FROM PLANT TISSUES, IN A PROCESS TERMED "CRYOTHERAPY". PLANT CRYOPROTECTANT FORMULATIONS HAVE REMAINED LARGELY UNCHANGED FOR THE PAST THREE DECADES; ALTHOUGH THEY HAVE BEEN USED SUCCESSFULLY, WE STILL DO NOT UNDERSTAND HOW OR WHY THESE FORMULATIONS WORK. THUS, IT IS CHALLENGING TO IMPROVE EXISTING FORMULATIONS OR PREDICT ALTERNATIVE FORMULATIONS THAT ARE EFFECTIVE FOR A WIDER RANGE OF GENEBANK ACCESSIONS.THIS GRANT WILL UTILIZE PHYSICAL CHEMICAL INSTRUMENTATION TO DIRECTLY MEASURE INTERACTIONS OF CRYOPROTECTANTS WITH THEIR SURROUNDINGS. STATE-OF-THE-ART COHERENT STIMULATED RAMAN MICROSCOPY WILL ALLOW US TO MEASURE THE LOCATION AND TRANSLOCATION OF CRYOPROTECTANT MOLECULES IN CULTURED PLANT CELL MODELS. IN PARALLEL, QUANTITATIVE MEASUREMENTS WITH ANALYTICAL SPECTROSCOPIES OF CRYOPROTECTANT FORMULATIONS AND CULTURED PLANT CELLS CORRELATED WITH RAMAN MICROSCOPY WILL REVEAL PROPERTIES AND INTERACTIONS OF THESE MOLECULES IN SITU IN CRYOPROTECTANT FORMULATIONS. RESULTS WILL REVEAL CRYOPROTECTANT INTERACTIONS WITH PLANT CELLS, LEAD TO THE DEVELOPMENT OF IMPROVED FORMULATION OF CRYOPROTECTANT SOLUTIONS, AND INCREASE OUR ABILITY TO USE CRYOTHERAPY TO ELIMINATE PLANT PATHOGENS, THUS LEADING TO HIGHLY EFFICIENT PLANT CRYOPRESERVATION AND CRYOTHERAPY PROCEDURES TO SECURE PLANT GENETIC RESOURCE COLLECTIONS.