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      Handheld tunable focus confocal microscope utilizing a double-clad fiber coupler for in vivo imaging of oral epithelium

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          Abstract.

          A reflectance confocal endomicroscope with double-clad fiber coupler and electrically tunable focus lens is applied to imaging of the oral mucosa. The instrument is designed to be lightweight and robust for clinical use. The tunable lens allows axial scanning through > 250    μ m in the epithelium when the probe tip is placed in contact with tissue. Images are acquired at 6.6 frames per second with a field of view diameter up to 850    μ m . In vivo imaging of a wide range of normal sites in the oral cavity demonstrates the accessibility of the handheld probe. In vivo imaging of clinical lesions diagnosed as inflammation and dysplasia illustrates the ability of reflectance confocal endomicroscopy to image cellular changes associated with pathology.

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          Most cited references40

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          In vivo confocal microscopy for diagnosis of melanoma and basal cell carcinoma using a two-step method: analysis of 710 consecutive clinically equivocal cases.

          We describe two algorithms to diagnose basal cell carcinomas (BCCs) and melanomas (MMs) using in vivo reflectance confocal microscopy (RCM). A total of 710 consecutive cutaneous lesions excised to exclude malignancy (216 MMs, 266 nevi, 119 BCCs, 67 pigmented facial macules, and 42 other skin tumors) were imaged by RCM. RCM features were correlated with pathology diagnosis to develop diagnostic algorithms. The diagnostic accuracy of the BCC algorithm defined on multivariate analysis of the training set (50%) and tested on the remaining cases was 100% sensitivity, 88.5% specificity. Positive features were polarized elongated features, telangiectasia and convoluted vessels, basaloid nodules, and epidermal shadowing corresponding to horizontal clefting. Negative features were non-visible papillae, disarrangement of the epidermal layer, and cerebriform nests. Multivariate discriminant analysis on the training set (excluding the BCCs) identified seven independently significant features for MM diagnosis. The diagnostic accuracy of the MM algorithm on the test set was 87.6% sensitivity, 70.8% specificity. The four invasive MMs that were misdiagnosed by RCM were all of nevoid subtype. RCM is a highly accurate non-invasive technique for BCC diagnosis. Good diagnostic accuracy was achieved also for MM diagnosis, although rare variants of melanocytic tumors may limit the strict application of the algorithm.
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            Confocal endomicroscopy: instrumentation and medical applications.

            Advances in fiber optic technology and miniaturized optics and mechanics have propelled confocal endomicroscopy into the clinical realm. This high resolution, non-invasive imaging technology provides the ability to microscopically evaluate cellular and sub-cellular features in tissue in vivo by optical sectioning. Because many cancers originate in epithelial tissues accessible by endoscopes, confocal endomicroscopy has been explored to detect regions of possible neoplasia at an earlier stage by imaging morphological features in vivo that are significant in histopathologic evaluation. This technique allows real-time assessment of tissue which may improve diagnostic yield by guiding biopsy. Research and development continues to reduce the overall size of the imaging probe, increase the image acquisition speed, and improve resolution and field of view of confocal endomicroscopes. Technical advances will continue to enable application to less accessible organs and more complex systems in the body. Lateral and axial resolutions down to 0.5 and 3 μm, respectively, field of view as large as 800 × 450 μm, and objective lens and total probe outer diameters down to 0.35 and 1.25 mm, respectively, have been achieved. We provide a review of the historical developments of confocal imaging in vivo, the evolution of endomicroscope instrumentation, and the medical applications of confocal endomicroscopy.
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              Light scattering from cervical cells throughout neoplastic progression: influence of nuclear morphology, DNA content, and chromatin texture.

              A number of noninvasive fiber optic optical technologies are under development for real-time diagnosis of neoplasia. We investigate how the light scattering properties of cervical cells are affected by changes in nuclear morphology, DNA content, and chromatin texture, which occur during neoplastic progression. We used a Cyto-Savant computer-assisted image analysis system to acquire quantitative nuclear features measurements from 122 Feulgen-thionin-stained histopathologic sections of cervical tissue. A subset of the measured nuclear features was incorporated into a finite-difference time-domain (FDTD) model of cellular light scattering. The magnitude and angular distribution of scattered light was calculated for cervical cells as a function of pathologic grade. The nuclear atypia strongly affected light scattering properties. The increased size and elevated DNA content of nuclei in high-grade lesions caused the most significant changes in scattering intensity. The spatial dimensions of chromatin texture features and the amplitude of refractive index fluctuations within the nucleus impacted both the angular distribution of scattering angles and the total amount of scattered light. Cellular scattering is sensitive to changes in nuclear morphology that accompany neoplastic progression. Understanding the quantitative relationships between nuclear features and scattering properties will aid in the development of noninvasive optical technologies for detection of precancerous conditions. Copyright 2003 Society of Photo-Optical Instrumentation Engineers
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                Author and article information

                Journal
                J Biomed Opt
                J Biomed Opt
                JBOPFO
                JBO
                Journal of Biomedical Optics
                Society of Photo-Optical Instrumentation Engineers
                1083-3668
                1560-2281
                25 May 2017
                May 2017
                : 22
                : 5
                : 056008
                Affiliations
                [a ]Texas A&M University , Biomedical Engineering Department, College Station, Texas, United States
                [b ]Texas A&M University College of Dentistry , Department of Diagnostic Sciences, Dallas, Texas, United States
                [c ]Texas A&M University College of Dentistry , Department of Periodontics, Dallas, Texas, United States
                Author notes
                [* ]Address all correspondence to: Kristen C. Maitland, E-mail: kmaitland@ 123456tamu.edu
                Article
                JBO-160863R 160863R
                10.1117/1.JBO.22.5.056008
                5444308
                28541447
                52e00177-a9e7-4638-b50c-0746add71306
                © The Authors.

                Published by SPIE under a Creative Commons Attribution 3.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

                History
                : 16 December 2016
                : 8 May 2017
                Page count
                Figures: 5, Tables: 0, References: 40, Pages: 8
                Funding
                Funded by: U.S. Department of Health and Human Services (HHS) http://dx.doi.org/10.13039/100000016
                Funded by: National Institutes of Health (NIH) http://dx.doi.org/10.13039/100000002
                Funded by: National Cancer Institute (NCI) http://dx.doi.org/10.13039/100000054
                Award ID: R01 CA138653
                Funded by: National Science Foundation (NSF) http://dx.doi.org/10.13039/100000001
                Award ID: 0750732
                Categories
                Research Papers: Imaging
                Paper
                Custom metadata
                Olsovsky et al.: Handheld tunable focus confocal microscope utilizing a double-clad fiber…

                Biomedical engineering
                confocal microscopy,microendoscopy,endomicroscopy,in vivo imaging,cancer
                Biomedical engineering
                confocal microscopy, microendoscopy, endomicroscopy, in vivo imaging, cancer

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