Multidepth imaging by chromatic dispersion confocal microscopy

Cory A. Olsovsky; Ryan L. Shelton; Meagan A. Saldua; Oscar Carrasco-Zevallos; Brian E. Applegate; Kristen C. Maitland

doi:10.1117/12.909175

28 February 2012 Multidepth imaging by chromatic dispersion confocal microscopy

Cory A. Olsovsky, Ryan L. Shelton, Meagan A. Saldua, Oscar Carrasco-Zevallos, Brian E. Applegate, Kristen C. Maitland

Proceedings Volume 8214, Advanced Biomedical and Clinical Diagnostic Systems X; 82140L (2012) https://doi.org/10.1117/12.909175
Event: SPIE BiOS, 2012, San Francisco, California, United States

Abstract

Confocal microscopy has shown potential as an imaging technique to detect precancer. Imaging cellular features throughout the depth of epithelial tissue may provide useful information for diagnosis. However, the current in vivo axial scanning techniques for confocal microscopy are cumbersome, time-consuming, and restrictive when attempting to reconstruct volumetric images acquired in breathing patients. Chromatic dispersion confocal microscopy (CDCM) exploits severe longitudinal chromatic aberration in the system to axially disperse light from a broadband source and, ultimately, spectrally encode high resolution images along the depth of the object. Hyperchromat lenses are designed to have severe and linear longitudinal chromatic aberration, but have not yet been used in confocal microscopy. We use a hyperchromat lens in a stage scanning confocal microscope to demonstrate the capability to simultaneously capture information at multiple depths without mechanical scanning. A photonic crystal fiber pumped with a 830nm wavelength Ti:Sapphire laser was used as a supercontinuum source, and a spectrometer was used as the detector. The chromatic aberration and magnification in the system give a focal shift of 140μm after the objective lens and an axial resolution of 5.2-7.6μm over the wavelength range from 585nm to 830nm. A 400x400x140μm³ volume of pig cheek epithelium was imaged in a single X-Y scan. Nuclei can be seen at several depths within the epithelium. The capability of this technique to achieve simultaneous high resolution confocal imaging at multiple depths may reduce imaging time and motion artifacts and enable volumetric reconstruction of in vivo confocal images of the epithelium.

Citation Download Citation

Cory A. Olsovsky, Ryan L. Shelton, Meagan A. Saldua, Oscar Carrasco-Zevallos, Brian E. Applegate, and Kristen C. Maitland "Multidepth imaging by chromatic dispersion confocal microscopy", Proc. SPIE 8214, Advanced Biomedical and Clinical Diagnostic Systems X, 82140L (28 February 2012); https://doi.org/10.1117/12.909175

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KEYWORDS

Confocal microscopy

Chromatic aberrations

Dispersion

Point spread functions

Lenses

Spectroscopy

Colorimetry

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