LINE SCAN FOCAL MODULATION MICROSCOPY FOR RAPID IMAGING OF THICK BIOLOGICAL SAMPLES

Abstract

LINE-SCAN FOCAL MODULATION MICROSCOPY IS A HIGH-SPEED, HIGH-RESOLUTION IMAGING TECHNIQUE CAPABLE OF DEEP IMAGING OF THICK BIOLOGICAL TISSUES. THIS THESIS DESCRIBES THREE DIFFERENT IMPLEMENTATIONS OF THE LINE-SCANNING FOCAL MODULATION MICROSCOPE. THE PERFORMANCE OF THE MICROSCOPE IS VALIDATED BY IMAGING FLUORESCENT BEADS EMBEDDED IN A SCATTERING GEL. THE SIGNAL-TO-BACKGROUND RATIO FOR THE LINE-SCAN FMM SYSTEM IS ABOUT 8 TIMES HIGHER THAN FOR LINE-SCANNING CONFOCAL SYSTEMS EVEN AT DEPTHS >100 ?M. THUS, THIS SYSTEM RETAINS ITS SECTIONING CAPABILITY EVEN AT GREAT DEPTHS. THE LINE-SCAN FMM SYSTEM HAS BEEN USED TO IMAGE THE BLOOD FLOW IN VARIOUS ZEBRAFISH MODELS. COMPARISON OF THE LINE-SCAN FMM IMAGE WITH DIFFERENT COMMERCIAL MICROSCOPES SHOWS THAT THE FMM SYSTEM HAS BETTER SPATIAL AND TEMPORAL RESOLUTION AND IS WELL SUITED FOR HIGH-SPEED IN VIVO IMAGING IN BIOLOGICAL TISSUES.