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. 2017 Jul 24;8(8):3807-3815.
doi: 10.1364/BOE.8.003807. eCollection 2017 Aug 1.

Line scanning, stage scanning confocal microscope (LSSSCM)

Daniel S Gareau  1 , James G Krueger  1 , Jason E Hawkes  1 , Samantha R Lish  1 , Michael P Dietz  1 , Alba Guembe Mülberger  1 , Euphemia W Mu  2 , Mary L Stevenson  2 , Jesse M Lewin  3 , Shane A Meehan  2 , John A Carucci  2
Affiliations

Line scanning, stage scanning confocal microscope (LSSSCM)

Daniel S Gareau et al. Biomed Opt Express. .

Abstract

For rapid pathological assessment of large surgical tissue excisions with cellular resolution, we present a line scanning, stage scanning confocal microscope (LSSSCM). LSSSCM uses no scanning mirrors. Laser light is focused with a single cylindrical lens to a line of diffraction-limited width directly into the (Z) sample focal plane, which is parallel to and near the flattened specimen surface. Semi-confocal optical sections are derived from the linear array distribution (Y) and a single mechanical drive that moves the sample parallel to the focal plane and perpendicular to the focused line (X). LSSSCM demonstrates cellular resolution in the conditions of high nuclear density within micronodular basal cell carcinoma.

Keywords: (110.0180) Microscopy; (170.1790) Confocal microscopy; (170.1870) Dermatology; (170.5810) Scanning microscopy; (170.7050) Turbid media.

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Conflict of interest statement

DSG: (P)

Figures

Fig. 1
Fig. 1
Cancer incidence rates in the United States. 5.4 million [3] of the 6 million new cancer cases in the US are the non-melanoma skin cancers that confocal point scanning systems have proven effective in detecting: Basal Cell Carcinoma (BCC) [8] and Squamous Cell Carcinoma (SCC). [9]
Fig. 2
Fig. 2
Stage-Scanning, Line Scanning Confocal Microscope in concept (a) and physical embodiment (b). The experimental result sample image (c) shows a 12-mm horizontal field of view scanned in 2 minutes without mosaicking. The sample has been cut into 4 pieces, stained (with various immersion times of 5, 10, 20 and 30 seconds) in 1mM acridine orange solution (pH=6.0), and re-assembled for imaging.
Fig. 3
Fig. 3
Image analysis of mirrored grating target. The image (a) is sampled along a single X or Y position to extract pixel values (b, black *) that comprise an edge trace which is fit using a sigmoidal function (b, blue line) that specifies the 10% and 90% brightness coordinates (b, red *).
Fig. 4
Fig. 4
Confocal image of basal cell carcinoma (BCC), acquired 10µm beneath the surface of a 4mm-thick tissue specimen with correlating histology. The inset shows a magnified view of a single tumor with characteristic pathologic features identified.
See this image and copyright information in PMC

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