Dataset Information

Real-time video mosaicking to guide handheld in vivo microscopy.

ABSTRACT: Handheld and endoscopic optical-sectioning microscopes are being developed for noninvasive screening and intraoperative consultation. Imaging a large extent of tissue is often desired, but miniature in vivo microscopes tend to suffer from limited fields of view. To extend the imaging field during clinical use, we have developed a real-time video mosaicking method, which allows users to efficiently survey larger areas of tissue. Here, we modified a previous post-processing mosaicking method so that real-time mosaicking is possible at >30 frames/second when using a device that outputs images that are 400 × 400 pixels in size. Unlike other real-time mosaicking methods, our strategy can accommodate image rotations and deformations that often occur during clinical use of a handheld microscope. We perform a feasibility study to demonstrate that the use of real-time mosaicking is necessary to enable efficient sampling of a desired imaging field when using a handheld dual-axis confocal microscope.

SUBMITTER: Yin C

PROVIDER: S-EPMC7969124 | biostudies-literature | 2020 Jun

REPOSITORIES: biostudies-literature

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Real-time video mosaicking to guide handheld in vivo microscopy.

Yin Chengbo C Wei Linpeng L Kose Kivanc K Glaser Adam K AK Peterson Gary G Rajadhyaksha Milind M Liu Jonathan T C JTC

Journal of biophotonics 20200414 6

Handheld and endoscopic optical-sectioning microscopes are being developed for noninvasive screening and intraoperative consultation. Imaging a large extent of tissue is often desired, but miniature in vivo microscopes tend to suffer from limited fields of view. To extend the imaging field during clinical use, we have developed a real-time video mosaicking method, which allows users to efficiently survey larger areas of tissue. Here, we modified a previous post-processing mosaicking method so th ...[more]

PMID: 32246558

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Project description:BACKGROUND:The surgical removal of non-melanoma skin cancers (NMSCs) is guided by the pathologic examination of margins. However, the preparation of histopathology is time consuming, labour-intensive and requires separate laboratory infrastructure. Furthermore, when histopathology indicates positive margins, patients must return for re-excisions. Reflectance confocal microscopy (RCM) with a new video-mosaicking approach can noninvasively delineate margins directly on patients and potentially guide surgery in real-time, augmenting the traditional approaches of histopathology. OBJECTIVE:To assess a new peri-operative RCM video-mosaicking approach for comprehensive delineation of NMSC margins on patients in vivo. METHODS:Thirty-five patients undergoing Mohs micrographic surgery (MMS) in the Mohs surgery unit at Memorial Sloan Kettering Cancer Center, New York, NY were included in the study. RCM imaging was performed before and after the first staged excision by acquiring videos along the surgical margins (epidermal, peripheral and deep dermal) of each wound, which were subsequently processed into video-mosaics. Two RCM evaluators read and assessed video-mosaics, and subsequently compared to the corresponding Mohs frozen histopathology. RESULTS:Reflectance confocal microscopy videos and video-mosaics displayed acceptable imaging quality (resolution and contrast), pre-operatively in 32/35 (91%) NMSC lesions and intra-operatively in 29/35 lesions (83%). Pre-operative delineation of margins correlated with the histopathology in 32/35 (91%) lesions. Intra-operative delineation correlated in 10/14 (71%) lesions for the presence of residual tumour and in 18/21 (86%) lesions for absence. Sensitivity/specificity were 71%/86% and 86%/81% for two RCM video-mosaic evaluators, and overall agreement was 80% and 83% with histopathology, with moderate inter-evaluator agreement (k = 0.59, P ≤ 0.0002). CONCLUSIONS:Peri-operative RCM video-mosaicking of NMSC margins directly on patients may potentially guide surgery in real-time, serve as an adjunct to histopathology, reduce time spent in clinic and reduce the need for re-excisions. Further testing in larger studies is needed.

Project description:Live-subject microscopies, including microendoscopy and other related technologies, offer promise for basic biology research as well as the optical biopsy of disease in the clinic. However, cellular resolution generally comes with the trade-off of a microscopic field-of-view. Microimage mosaicking enables stitching many small scenes together to aid visualization, quantitative interpretation, and mapping of microscale features, for example, to guide surgical intervention. The development of hyperspectral and multispectral systems for biomedical applications provides motivation for adapting mosaicking algorithms to process a number of simultaneous spectral channels. We present an algorithm that mosaics multichannel video by correlating channels of consecutive frames as a basis for efficiently calculating image alignments. We characterize the noise tolerance of the algorithm by using simulated video with known ground-truth alignments to quantify mosaicking accuracy and speed, showing that multiplexed molecular imaging enhances mosaic accuracy by leveraging observations of distinct molecular constituents to inform frame alignment. A simple mathematical model is introduced to characterize the noise suppression provided by a given group of spectral channels, thus predicting the performance of selected subsets of data channels in order to balance mosaic computation accuracy and speed. The characteristic noise tolerance of a given number of channels is shown to improve through selection of an optimal subset of channels that maximizes this model. We also demonstrate that the multichannel algorithm produces higher quality mosaics than the analogous single-channel methods in an empirical test case. To compensate for the increased data rate of hyperspectral video compared to single-channel systems, we employ parallel processing via GPUs to alleviate computational bottlenecks and to achieve real-time mosaicking even for video-rate multichannel systems anticipated in the future. This implementation paves the way for real-time multichannel mosaicking to accompany next-generation hyperspectral and multispectral video microscopy.

Dataset Information

Real-time video mosaicking to guide handheld in vivo microscopy.

Publications

Real-time video mosaicking to guide handheld in vivo microscopy.

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