Strip mosaicing confocal microscopy for rapid imaging over large areas of excised tissue

Abeytunge, Sanjeewa; Li, Yongbiao; Larson, Bjorg A.; Peterson, Gary; Toledo-Crow, Ricardo; Rajadhyaksha, Milind

doi:10.1117/12.909416

Cited by 2 publications

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“…Fluorescence confocal microscopy (FCM) has been previously described as a fast imaging tool that offers some advantages over frozen section analysis, such as shorter turnaround time and easy detection of adipose tissue, and its role in Mohs surgery has already been investigated. [20][21][22][23][24][25][26][27][28][29][30][31][32] However, studies addressing its accuracy on larger series are lacking.…”

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confidence: 99%

“…[1][2][3][4][5][6][7][8][9][10][11][12][13][14][15]20,[33][34][35][36][37][38] It was initially applied to skin cancer, in particular BCCs, and later to solid tumours in pivotal studies. [21][22][23][24][25][26][27][28][29][30][31][32] Basically, FCM scans a given surgical specimen and provides grey-scale high-resolution images that are automatically stitched together to create 'mosaics'. Each mosaic displays large areas of tissue covering the whole specimen area, as necessary for surgical pathology.…”

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confidence: 99%

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Diagnostic accuracy of ex vivo fluorescence confocal microscopy in Mohs surgery of basal cell carcinomas: a prospective study on 753 margins

et al. 2019

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Summary Background Frozen histological sections are used for intraoperative margin assessment during Mohs surgery. Fluorescence confocal microscopy (FCM) is a new tool that offers a promising and faster alternative to frozen histology. Objectives To evaluate prospectively in a clinical setting the accuracy of FCM vs. frozen sections in margin assessment of basal cell carcinoma (BCC). Methods Patients with BCC scheduled for Mohs surgery were prospectively enrolled. Freshly excised surgical specimens were examined by FCM and then frozen sections were evaluated. Permanent sections were obtained, in order to validate the sample technique. A blind re‐evaluation was also performed for discordant cases. Sensitivity and specificity levels, as well as positive and negative predictive values (PPV and NPV, respectively), were calculated and receiver–operating characteristic curves generated. Results We enrolled 127 BCCs in as many patients (40·2% females). Seven hundred and fifty‐three sections were examined. All BCCs were located in the head and neck area. In evaluating the performance of FCM vs. frozen sections, sensitivity was 79·8%, specificity was 95·8%, PPV was 80·5% and NPV was 95·7% [area under the curve 0·88, 95% confidence interval 0·84–0·92 (P < 0·001)]. Forty‐nine discordant cases were re‐evaluated; 24 were false positive and 25 false negative. The performance of FCM and frozen sections was also evaluated according to the final histopathological assessment. Conclusions We found high levels of accuracy for FCM vs. frozen section evaluation in intraoperative BCC margin assessment during Mohs surgery. Some technical issues prevent the wide use of this technique, but new devices promise to overcome these limitations.

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Diagnostic accuracy of ex vivo fluorescence confocal microscopy in Mohs surgery of basal cell carcinomas: a prospective study on 753 margins

et al. 2019

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“…Point scanning technology is able to scan faster (0.56mm 2 per second: 10mm × 10mm in 180 seconds) in a research version of the device using point-scanning, strip mosaicking mode. [24] The fastest reported scan rate is from a similar rapid line-scanning system (0.88mm 2 per second: 10mm × 10mm in 88 seconds) [20] using a 40-times, water immersion, 0.8 NA objective lens and the same 488nm LASER wavelength. In their system, 1 Airy unit is 0.74 µm so the illumination point spread function (PSF) is superior to (Eq.…”

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confidence: 99%

“…The PSF full width at half maximum in the X and Y directions are 0.47 µm ± 0.02 µm and 0.56 µm ± 0.01 µm(n = 5) in their system, respectively. [24] 10mm×10mm (180s) 0.56mm 2 per second Point Scanner [26] 5mm×5mm (84s) 0.30mm 2 per second An additional factor impacting utility and overall procedure speed is an automated adjustment of sample tip and tilt. This must be done precisely (5µm tip/tilt tolerance over the lateral 3cm field of view) and currently requires 5 minutes using manual micrometer adjustment and feedback from live line readouts.…”

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confidence: 99%

Line scanning, stage scanning confocal microscope (LSSSCM)

Gareau

Krueger

Hawkes

et al. 2017

Biomed. Opt. Express

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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.

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Strip mosaicing confocal microscopy for rapid imaging over large areas of excised tissue

Cited by 2 publications

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Diagnostic accuracy of ex vivo fluorescence confocal microscopy in Mohs surgery of basal cell carcinomas: a prospective study on 753 margins

Diagnostic accuracy of ex vivo fluorescence confocal microscopy in Mohs surgery of basal cell carcinomas: a prospective study on 753 margins

Line scanning, stage scanning confocal microscope (LSSSCM)

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