Rapid confocal imaging of large areas of excised tissue with strip mosaicing

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

doi:10.1117/1.3582335

Cited by 47 publications

(48 citation statements)

References 11 publications

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“…2 and point the reader to our previous publications for a more detailed description. 24,25 The basic idea of a mosaicking microscope is to generate a mosaic from individual high-resolution images. The CSM microscope acquires long strip images and stitches the image strips to complete the mosaic.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of breast tissue with confocal strip-mosaicking microscopy: a test approach emulating pathology-like examination

et al. 2017

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Abstract. Confocal microscopy is an emerging technology for rapid imaging of freshly excised tissue without the need for frozen-or fixed-section processing. Initial studies have described imaging of breast tissue using fluorescence confocal microscopy with small regions of interest, typically 750 × 750 μm 2 . We present exploration with a microscope, termed confocal strip-mosaicking microscope (CSM microscope), which images an area of 2 × 2 cm 2 of tissue with cellular-level resolution in 10 min of excision. Using the CSM microscope, we imaged 34 fresh, human, large breast tissue specimens from 18 patients, blindly analyzed by a board-certified pathologist and subsequently correlated with the corresponding standard fixed histopathology. Invasive tumors and benign tissue were clearly identified in CSM strip-mosaic images. Thirty specimens were concordant for image-to-histopathology correlation while four were discordant.

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Section: Introductionmentioning

confidence: 99%

Evaluation of breast tissue with confocal strip-mosaicking microscopy: a test approach emulating pathology-like examination

et al. 2017

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“…The sample is physically scanned perpendicular to the stationary line in the focal plane to acquire long rectangular images are stitched together in less time than square images. 13 The long rectangular images conveniently have no field curvature in their long (mechanically scanned) axis. These are rapid techniques compared to histology, which takes 20 to 45 min.…”

Section: Introductionmentioning

confidence: 99%

“…A recently developed "strip mosaicing" technique is even faster, producing a 10 × 10 mm mosaic in about 3 min. 13 In strip mosaicing, the slow-axis scanning galvanometer is fixed on the optical axis while the fast polygon scans a stationary line in the tissue. The sample is physically scanned perpendicular to the stationary line in the focal plane to acquire long rectangular images are stitched together in less time than square images.…”

Section: Introductionmentioning

confidence: 99%

Tri-modal confocal mosaics detect residual invasive squamous cell carcinoma in Mohs surgical excisions

et al. 2012

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Abstract. For rapid, intra-operative pathological margin assessment to guide staged cancer excisions, multimodal confocal mosaic scan image wide surgical margins (approximately 1 cm) with sub-cellular resolution and mimic the appearance of conventional hematoxylin and eosin histopathology (H&E). The goal of this work is to combine three confocal imaging modes: acridine orange fluorescence (AO) for labeling nuclei, eosin fluorescence (Eo) for labeling cytoplasm, and endogenous reflectance (R) for marking collagen and keratin. Absorption contrast is achieved by alternating the excitation wavelength: 488 nm (AO fluorescence) and 532 nm (Eo fluorescence). Superposition and false-coloring of these modes mimics H&E, enabling detection of cutaneous squamous cell carcinomas (SCC). The sum of mosaic Eo þ R is false-colored pink to mimic the appearance of eosin, while the AO mosaic is false-colored purple to mimic the appearance of hematoxylin in H&E. In this study, mosaics of 10 Mohs surgical excisions containing invasive SCC, and five containing only normal tissue were subdivided for digital presentation equivalent to 4× histology. Of the total 50 SCC and 25 normal sub-mosaics presented, two reviewers made two and three type-2 errors (false positives), respectively. Limitations to precisely mimic H&E included occasional elastin staining by AO. These results suggest that confocal mosaics may effectively guide staged SCC excisions in skin and other tissues.

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“…For our study, this added an additional ~1 second to the acquisition of each volumetric data set. An alternative to acquiring individual square fields is to obtain long strip images by continuously translating along one axis while acquiring the image [36]. In this case the acquisition speed for the total mosaic will be mainly limited by the A-scan rate of the system.…”

Section: Resultsmentioning

confidence: 99%

Swept source optical coherence microscopy using a 1310 nm VCSEL light source

et al. 2013

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Abstract:We demonstrate high speed, swept source optical coherence microscopy (OCM) using a MEMS tunable vertical cavity surface-emitting laser (VCSEL) light source. The light source had a sweep rate of 280 kHz, providing a bidirectional axial scan rate of 560 kHz. The sweep bandwidth was 117 nm centered at 1310 nm, corresponding to an axial resolution of 13.1 µm in air, corresponding to 8.1 µm (9.6 µm spectrally shaped) in tissue. Dispersion mismatch from different objectives was compensated numerically, enabling magnification and field of view to be easily changed. OCM images were acquired with transverse resolutions between 0.86 µm -3.42 µm using interchangeable 40X, 20X and 10X objectives with ~600 µm x 600 µm, ~1 mm x 1 mm and ~2 mm x 2 mm field-of-view (FOV), respectively. Parasitic variations in path length with beam scanning were corrected numerically. These features enable swept source OCM to be integrated with a wide range of existing scanning microscopes. Large FOV mosaics were generated by serially acquiring adjacent overlapping microscopic fields and combining them in post-processing. Fresh human colon, thyroid and kidney specimens were imaged ex vivo and compared to matching histology sections, demonstrating the ability of OCM to image tissue specimens.

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Rapid confocal imaging of large areas of excised tissue with strip mosaicing

Cited by 47 publications

References 11 publications

Evaluation of breast tissue with confocal strip-mosaicking microscopy: a test approach emulating pathology-like examination

Evaluation of breast tissue with confocal strip-mosaicking microscopy: a test approach emulating pathology-like examination

Tri-modal confocal mosaics detect residual invasive squamous cell carcinoma in Mohs surgical excisions

Swept source optical coherence microscopy using a 1310 nm VCSEL light source

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