Multimodal optical microscope for detecting viability of mouse embryos in vitro

• Published in: Journal of biomedical optics 2007-07, Vol.12 (4), p.044006-044006
• Main Authors: Warger, 2nd, William C, Laevsky, Gary S, Townsend, Daniel J, Rajadhyaksha, Milind, DiMarzio, Charles A
• Format: Article
• Language: English
• Subjects: Animals; Confocal; Embryo, Mammalian - cytology; Embryo, Mammalian - physiology; Embryos; Equipment Design; Equipment Failure Analysis; Fetal Viability - physiology; Illumination; Image Enhancement - instrumentation; Image Enhancement - methods; Image Interpretation, Computer-Assisted - instrumentation; Image Interpretation, Computer-Assisted - methods; Imaging; Mice; Microscopy; Microscopy, Confocal - instrumentation; Microscopy, Confocal - methods; Microscopy, Fluorescence - instrumentation; Microscopy, Fluorescence - methods; Microscopy, Polarization - instrumentation; Microscopy, Polarization - methods; Optical microscopes; Scanning; Systems Integration
• ISSN: 1083-3668
• DOI: 10.1117/1.2753744

We present a multimodal optical microscope that incorporates six imaging modalities on one common platform. The imaging modalities include three staring modes, optical quadrature microscopy (OQM), differential interference contrast (DIC) microscopy, and epi-fluorescence microscopy, and three scanning modes, confocal reflectance microscopy (CRM), confocal fluorescence microscopy (CFM), and two-photon microscopy (2PM). OQM reconstructs the amplitude and phase of an optically transparent specimen within a modified Mach-Zehnder configuration. DIC microscopy images the phase gradient along a specified direction of an optically transparent specimen. CRM detects index of refraction changes that modulate backscatter. Epi-fluorescence microscopy, CFM, and 2PM detect endogenous and exogenous fluorophores within a specimen. The scanning modes are inherently capable of producing three-dimensional (3-D) images due to optical sectioning and localized probing. Illumination and imaging are performed coaxially with minimal changes of optical components between modes. Multimodal images of embryos are shown to demonstrate the microscope's imaging capabilities.