Full pupil line-scanning confocal microscope for imaging weakly scattering tissues: comparison to divided pupil

Daniel S. Gareau; Sanjee Abeytunge; Milind Rajadhyaksha

doi:10.1117/12.809450

12 February 2009 Full pupil line-scanning confocal microscope for imaging weakly scattering tissues: comparison to divided pupil

Daniel S. Gareau, Sanjee Abeytunge, Milind Rajadhyaksha

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Proceedings Volume 7182, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VII; 718207 (2009) https://doi.org/10.1117/12.809450
Event: SPIE BiOS, 2009, San Jose, California, United States

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 μm, which degrades through fullthickness human epidermis to 2.8 +/- 0.78 μm. The lateral resolution is 0.7±0.10 μm, which degrades to 1.6±0.28 μm through human epidermis. The divided-pupil design demonstrates instrumental optical sectioning of 1.7 μm, which degrades to 7.6 μm through human epidermis. The lateral resolution is 1.0 μm, which degrades to 1.7 μm. 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.

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Daniel S. Gareau, Sanjee Abeytunge, and Milind Rajadhyaksha "Full pupil line-scanning confocal microscope for imaging weakly scattering tissues: comparison to divided pupil", Proc. SPIE 7182, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VII, 718207 (12 February 2009); https://doi.org/10.1117/12.809450

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KEYWORDS

Scattering

Confocal microscopy

Tissues

Microscopes

Tissue optics

Skin

In vivo imaging

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