Raman spectroscopic analysis of pancreatic islets.

RAMAN SPECTROSCOPIC ANALYSIS OF PANCREATIC ISLETS Kimberly Hobbs, Emanuel Waddell, Adriel D. Johnson, Sr., University of Alabama in Huntsville, Huntsville, Alabama 35899. Jacqueline U. Johnson, Alabama A & M University, Normal, Alabama, 35762.

Raman spectroscopy can be used as a method for characterizing chemical properties of cells. Our studies explore the feasibility of Raman spectroscopy in differentiation and functional studies of pancreatic islet cells. Alloxan monohydrate was used to induce hyperglycemia in Sprague Dawley rats. Islets were isolated using a liberase enzyme prepared in HBSS buffer which was injected into the common bile duct to distend the pancreas. The distended pancreas was placed in a conical tube containing the liberase solution for tissue digestion. The pancreatic dispersion was removed and islet cells were purified using a Ficoll Purification gradient. Islets were handpicked using an inverted surgical microscope and plated in DMEM. Islet specificity was determined with dithizone (DTZ) stain, which binds to the Zn2+ molecule of insulin revealing a red color. Photomicrographs were taken of freshly isolated islets, alloxan treated islets, and the DTZ stained islets. Islets were placed on an excimer laser optic chip, examined using a Horiba Jobin Yvon HR 800 Confocal Raman Spectrometer, and scanned using a He-Ne 635 nm laser. Preliminary results using Raman spectroscopy revealed a significant peak analysis at a Raman shift of approximately 3000 cm-1. To our knowledge Raman spectroscopy has never been used to examine pancreatic islets under these conditions. These studies may lead to the discovery of novel information about pancreatic function and a better understanding of conditions such as diabetes.