Confocal reflectance theta line scanning microscope for imaging human skin in vivo

Institute of Optics, University of Rochester, Rochester, New York, United States
Optics Letters (Impact Factor: 3.18). 05/2006; 31(7):942-4. DOI: 10.1364/OL.31.000942
Source: PubMed

ABSTRACT A confocal reflectance theta line scanning microscope demonstrates imaging of nuclear and cellular detail in human epidermis in vivo. Experimentally measured line-spread functions determine the instrumental optical section thickness to be 1.7 +/- 0.1 microm and the lateral resolution to be 1.0 +/- 0.1 microm. Within human dermis (through full-thickness epidermis), the measured section thickness is 9.2 +/- 1.7 microm and the lateral resolution is 1.7 +/- 0.1 microm. An illumination line is scanned directly in the pupil of the objective lens, and the backscattered descanned light is detected with a linear array, such that the theta line scanner consists of only seven optical components.

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    • "Optical coherence microscopy (OCM) combines the high-sensitivity coherence gate of OCT and high transverse resolution of confocal microscopy to achieve cellular level imaging of tissue microstructures in 3-D [17], [19], [22], [91]. Confocal microscopy can detect either the reflectance (scattered) light [92]–[94] or the fluorescence light [95]–[97]. Multiphoton microscopy (MPM) utilizes the nonlinear generation of fluorescence from endogenous and exogenous markers in the tissue [10], [98], [99]. "
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    • "Figure 1 shows the preliminary experimental results form the full-pupil design. By comparison, a divided-pupil confocal line-scanning microscope demonstrates nominal instrumental optical sectioning of 1.7 μm which degrades to 7.6 μm through human epidermis [4] . "
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    ABSTRACT: Confocal reflectance full-pupil and divided-pupil line-scanning microscopes provide optical sectioning and image nuclear detail in skin. Line-scanning with linear detectors is a simpler alternative to point-scanning for imaging weakly scattering epidermis and the oral epithelium. With illumination of 830 nm, a water immersion lens of numerical aperture 0.9 and slit width three times smaller than the diffraction-limited line width, the instrumental full width at half maximum (FWHM) optical sectioning (linespread function) for the full-pupil design is 1.4 +/- 0.07 mum, which degrades through fullthickness human epidermis to 2.8 +/- 0.78 mum. The lateral resolution is 0.7+/-0.10 mum, which degrades to 1.6+/-0.28 mum through human epidermis. The divided-pupil design demonstrates instrumental optical sectioning of 1.7 mum, which degrades to 7.6 mum through human epidermis. The lateral resolution is 1.0 mum, which degrades to 1.7 mum. Heavy scattering in the dermis decreases contrast. Images of skin in-vivo show nuclear detail as expected with the predicted and experimentally verified sectioning. However, pixel crosstalk and speckle artifact degrade image quality in strongly scattering and aberrating tissues. The sources of degradation (aberration and scattering) are evaluated for the two design to assess the feasibility of these techniques for in vivo imaging.
    Proceedings of SPIE - The International Society for Optical Engineering 10/2008; DOI:10.1117/12.809450 · 0.20 Impact Factor
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    • "A further variant is reflectance confocal which uses reflected rather than fluorescence light to perform confocal sectioning. Reflectance confocal is frequently used in clinical and preclinical applications (see, e.g., Collier et al., 2007; Dwyer et al., 2006; González and Tannous, 2002). More recently, spectral scanning has been employed to enable finer spectral resolution of signals emanating from the sample (reviewed in Dickinson et al., 2001; Zimmermann et al., 2003). "
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