Wide-field imaging combined with confocal microscopy using a miniature f/5 camera integrated within a high NA objective lens

Dickensheets, David L.; Kreitinger, Seth; Peterson, Gary; Heger, Michael; Rajadhyaksha, Milind

doi:10.1364/ol.42.001241

Cited by 11 publications

(6 citation statements)

References 9 publications

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“…In the future, we will integrate an additional miniature camera that images through the same objective lens as CPM but with a significantly smaller NA and larger field to acquire wide-field dermatoscopy images spatially registered with CPM images, similarly to previous work on integrating dermatoscopy as part of an RCM device. 24 Additional microscopic objects were observable other than melanocytic cells and capillaries. For example, a white object was noticed at the centre of Figure 7C, D and F.…”

Section: Discussionmentioning

confidence: 99%

Handheld cross‐polarised microscope for imaging individual pigmented cells in human skin in vivo

Romero,

Zhao,

Stratton

et al. 2023

Journal of Microscopy

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We present the development of a simple, handheld cross‐polarised microscope (CPM) and demonstration of imaging individual pigmented cells in human skin in vivo. In the CPM device, the cross‐polarised detection approach is used to reduce the specular reflection from the skin surface and preferentially detect multiply‐scattered light. The multiply‐scattered light works as back illumination from within the tissue towards the skin surface, and superficial pigment such as intraepidermal melanin absorbs some spectral bands of the multiply‐scattered light and cast coloured shadows. Since the light that interacted with the superficial pigment only needs to travel a short distance before it exits the skin surface, microscopic details of the pigment can be preserved. The CPM device uses a water‐immersion objective lens with a high numerical aperture to image the microscopic details with minimal spherical aberrations and a small depth of focus. Preliminary results from a pilot study of imaging skin lesions in vivo showed that the CPM device could reveal three‐dimensional distribution of pigmented cells and intracellular distribution of pigment. Co‐registered CPM and reflectance confocal microscopy images showed good correspondence between dark, brown cells in CPM images and bright, melanin‐containing cells in reflectance confocal microscopy images.

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Section: Discussionmentioning

confidence: 99%

Handheld cross‐polarised microscope for imaging individual pigmented cells in human skin in vivo

Romero,

Zhao,

Stratton

et al. 2023

Journal of Microscopy

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“…Acquiring both multiple video‐mosaics and depth‐stacks will facilitate probing of a large volume of tissue in vivo . RCM imaging may also benefit from integration with widefield approaches such as with an embedded microcamera , or with fluorescence lifetime imaging microscopy or autofluorescence . An initial widefield‐of‐view examination may quickly guide subsequent RCM imaging in focal areas of suspicion.…”

Section: Discussionmentioning

confidence: 99%

Feasibility of a Video‐Mosaicking Approach to Extend the Field‐of‐View For Reflectance Confocal Microscopy in the Oral Cavity In Vivo

et al. 2019

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Background Reflectance confocal microscopy (RCM) is a developing approach for noninvasive detection of oral lesions with label‐free contrast and cellular‐level resolution. For access into the oral cavity, confocal microscopes are being configured with small‐diameter telescopic probes and small objective lenses. However, a small probe and objective lens allows for a rather small field‐of‐view relative to the large areas of tissue that must be examined for diagnosis. To extend the field‐of‐view for intraoral RCM imaging, we are investigating a video‐mosaicking approach. Methods A relay telescope and objective lens were adapted to an existing confocal microscope for access into the oral cavity. Imaging was performed using metal three‐dimensional‐printed objective lens front‐end caps with coverslip windows to contact and stabilize the tissue and set depth. Four healthy volunteers (normal oral mucosa), one patient (with an amalgam tattoo) in a clinical setting, and 20 anesthetized patients (with oral squamous cell carcinoma [OSCC]) in a surgical setting were imaged. Instead of the usual still RCM images, videos were recorded and then processed into video‐mosaics. Thirty video‐mosaics were read and qualitatively assessed by an expert reader of RCM images of the oral mucosa. Results Whereas the objective lens’ native field‐of‐view is 0.75 mm × 0.75 mm, the video‐mosaics display larger areas, ranging from 2 mm × 2 mm to 4 mm × 2 mm, with resolution, morphologic detail, and image quality that is preserved relative to that observed in the original videos (individual images). Video‐mosaics in healthy volunteers’ and the patients’ images showed cellular morphologic patterns in the lower epithelium and at the epithelial junction, and connective tissue along with capillary loops and blood flow in the deeper lamina propria. In OSCC, tumor nests could be observed along with normal looking mucosa in margin areas. Conclusions Video‐mosaicking is a reasonably quick and efficient approach for extending the field‐of‐view of RCM imaging, which can, to some extent, overcome the inherent limitation of an intraoral probe's small field‐of‐view. Reading video‐mosaics can mimic the procedure for examining pathology: initial visualization of the spatial cellular and morphologic patterns of the tumor and the spread of tumor margins over larger areas of the lesion, followed by digitally zooming (magnifying) for closer inspection of suspicious areas. However, faster processing of videos into video‐mosaics will be necessary, to allow examination of video‐mosaics in real‐time at the bedside. Lasers Surg. Med. 51:439–451, 2019. © 2019 Wiley Periodicals, Inc.

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“… 24 In another example, for skin-cancer, a micro-camera for wide-field imaging has been integrated into the objective lens of an RCM device, providing a

field of view to navigate the positioning of a confocal microscope with a field of view of

. 103 …”

Section: Opportunities and Best Practicesmentioning

confidence: 99%

In vivo microscopy as an adjunctive tool to guide detection, diagnosis, and treatment

et al. 2022

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Wide-field imaging combined with confocal microscopy using a miniature f/5 camera integrated within a high NA objective lens

Cited by 11 publications

References 9 publications

Handheld cross‐polarised microscope for imaging individual pigmented cells in human skin in vivo

Handheld cross‐polarised microscope for imaging individual pigmented cells in human skin in vivo

Feasibility of a Video‐Mosaicking Approach to Extend the Field‐of‐View For Reflectance Confocal Microscopy in the Oral Cavity In Vivo

In vivo microscopy as an adjunctive tool to guide detection, diagnosis, and treatment

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