Development and optimization of a line-scanned dual-axis confocal (LS-DAC) microscope for high-speed pathology

We have developed a line-scanned dual-axis confocal (LS-DAC) microscope with subcellular resolution suitable for real time diagnostic imaging at shallow depths. This design serves as a benchtop prototype for a handheld version of the LS-DAC intended for rapid point-of-care pathology. We have assessed the performance trade-offs between the LS-DAC and a point-scanned dual-axis confocal (PS-DAC) microscope via diffraction-theory analysis, Monte-Carlo simulations, and characterization experiments with phantoms and fresh tissues. In addition, we are exploring the use of a sCMOS detector array and rapid 3D deconvolution to improve the sensitivity and resolution of our LS-DAC microscope.

[1]  R. Webb,et al.  Video-rate confocal scanning laser microscope for imaging human tissues in vivo. , 1999, Applied optics.

[2]  Joel N. Bixler,et al.  Confocal Endomicroscopy: Instrumentation and Medical Applications , 2012, Annals of Biomedical Engineering.

[3]  G S Kino,et al.  Micromachined scanning confocal optical microscope. , 1996, Optics letters.

[4]  Y. Wang,et al.  Comparison of line-scanned and point-scanned dual-axis confocal microscope performance. , 2013, Optics letters.

[5]  Richard M. Levenson,et al.  Point-of-Care Pathology with Miniature Microscopes , 2011, Analytical cellular pathology.

[6]  B Bouma,et al.  A GRISM-based probe for spectrally encoded confocal microscopy. , 2003, Optics express.

[7]  Jonathan T. C. Liu,et al.  Assessing the tissue-imaging performance of confocal microscope architectures via Monte Carlo simulations. , 2012, Optics letters.

[8]  Y. Sabharwal,et al.  Slit-scanning confocal microendoscope for high-resolution in vivo imaging. , 1999, Applied optics.

[9]  Nathan O. Loewke,et al.  Micromirror-scanned dual-axis confocal microscope utilizing a gradient-index relay lens for image guidance during brain surgery. , 2010, Journal of biomedical optics.

[10]  Robert T Kester,et al.  Low cost, high performance, self-aligning miniature optical systems. , 2009, Applied optics.

[11]  Matthew D. Chidley,et al.  In vivo fiber-optic confocal reflectance microscope with an injection-molded plastic miniature objective lens. , 2005, Applied optics.

[12]  Ye Chen,et al.  Optimizing the performance of dual-axis confocal microscopes via Monte-Carlo scattering simulations and diffraction theory , 2013, Journal of biomedical optics.

[13]  Thomas D. Wang,et al.  Efficient rejection of scattered light enables deep optical sectioning in turbid media with low-numerical-aperture optics in a dual-axis confocal architecture. , 2008, Journal of biomedical optics.