Ultra-slim plastic endomicroscope objective for non-linear microscopy.

Non-linear microscopy has the potential to provide clinically useful information on the structure of biological tissue in vivo via an endomicroscope. The ability to use plastic as the optical material in a multiphoton objective was evaluated based on several criteria including autofluorescence, injection molding induced birefringence, and pulse broadening due to group velocity dispersion. An all-plastic, refractive ultra-slim endoscope objective was built with design specifications of NA=0.

Hyperspectral Shack-Hartmann test.

A hyperspectral Shack-Hartmann test bed has been developed to characterize the performance of miniature optics across a wide spectral range, a necessary first step in developing broadband achromatized all-polymer endomicroscopes. The Shack-Hartmann test bed was used to measure the chromatic focal shift (CFS) of a glass singlet lens and a glass achromatic lens, i.e.

Design and evaluation of an ultra-slim objective for in-vivo deep optical biopsy.

An estimated 1.6 million breast biopsies are performed in the US each year. In order to provide real-time, in-vivo imaging with sub-cellular resolution for optical biopsies, we have designed an ultra-slim objective to fit inside the 1-mm-diameter hypodermic needles currently used for breast biopsies to image tissue stained by the fluorescent probe proflavine.

Imaging performance of a miniature integrated microendoscope.

An integrated miniature multi-modal microscope (4M device) for microendoscopy was built and tested. Imaging performance is evaluated and imaging results are presented for both fluorescence and reflectance samples. Images of biological samples show successful imaging of both thin layers of fixed cells prepared on a slide as well as thick samples of excised fixed porcine epithelial tissue, thus demonstrating the potential for in vivo use.

In vivo imaging of oral neoplasia using a miniaturized fiber optic confocal reflectance microscope.

The purpose of this study was to determine whether in vivo images of oral mucosa obtained with a fiber optic confocal reflectance microscope could be used to differentiate normal and neoplastic tissues. We imaged 20 oral sites in eight patients undergoing surgery for squamous cell carcinoma. Normal and abnormal areas within the oral cavity were identified clinically, and real-time videos of each site were obtained in vivo using a fiber optic confocal reflectance microscope.

High numerical aperture microendoscope objective for a fiber confocal reflectance microscope.

A disposable high numerical aperture microendoscope objective has been designed, fabricated, and tested for use with a fiber confocal reflectance microscope. The objective uses high precision LIGA fabricated components to integrate imaging components and hydraulic suction lines into a housing that measures only 3.85 mm in outer diameter and 14.

Molecular optical imaging of therapeutic targets of cancer.

Recent progress in discerning the molecular events that accompany carcinogenesis has led to development of new cancer therapies directly targeted against the molecular changes of neoplasia. Molecular-targeted therapeutics have shown significant improvements in response rates and decreased toxicity as compared to conventional cytotoxic therapies which lack specificity for tumor cells. In order to fully explore the potential of molecular-targeted therapy, a new set of tools is required to dynamically and quantitatively image and monitor the heterogeneous molecular profiles of tumors in vivo.

Eye-movement study and human performance using telepathology virtual slides: implications for medical education and differences with experience.

A core skill in diagnostic pathology is light microscopy. Remarkably little is known about human factors that affect the proficiency of pathologists as light microscopists. The cognitive skills of pathologists have received relatively little attention in comparison with the large literature on human performance studies in radiology.

Design, assembly, and optical bench testing of a high-numerical-aperture miniature injection-molded objective for fiber-optic confocal reflectance microscopy.

The design, analysis, assembly methods, and optical-bench test results for a miniature injection-molded plastic objective lens used in a fiber-optic confocal reflectance microscope are presented. The five-lens plastic objective was tested as a stand-alone optical system before its integration into a confocal microscope for in vivo imaging of cells and tissue. Changing the spacing and rotation of the individual optical elements can compensate for fabrication inaccuracies and improve performance.

Removal of ghost images by using tilted element optical systems with polynomial surfaces for aberration compensation.

A novel solution to problematic ghost images is implemented by using tilted lens elements with polynomial surfaces. Tilting the lens surfaces sends reflections out of the imaging path. The nonrotationally symmetric polynomial surfaces correct aberrations caused by tilts.

The evolution of scanning microphotometry: a historical review.

This article traces the evolution of scanning microphotometry from its beginning, with an emphasis on the developing technologies that have made feasible the rapid and detailed capture of high-resolution image information. Consideration is given to future directions that may prove fruitful to clinical practice.

Optical imaging of cervical pre-cancers with structured illumination: an integrated approach.

Objective: Structured illumination microscopy is an inexpensive alternative to confocal microscopy that allows optical sectioning at a sub-cellular resolution. However, its application in imaging biological tissue has been limited by inadequate contrast present in them especially in reflectance imaging. Novel, optically active contrast agents like gold nanoparticles and quantum dots targeted against biomarkers of cancer can be integrated with structured illumination to image both the morphological and biochemical changes associated with epithelial pre-cancers.

Confocal microscopy: imaging cervical precancerous lesions.

Objectives: We explore the clinical potential of reflectance and fluorescence confocal microscopy to image the morphologic and biochemical changes associated with precancer, in order to aid in the detection and diagnosis of cervical dysplasia.

Methods: Cervical epithelial tissue samples imaged ex vivo or in vivo were obtained from M. D.

Sorting method to extend the dynamic range of the Shack-Hartmann wave-front sensor.

We propose a simple and powerful algorithm to extend the dynamic range of a Shack-Hartmann wave-front sensor. In a conventional Shack-Hartmann wave-front sensor the dynamic range is limited by the f-number of a lenslet, because the focal spot is required to remain in the area confined by the single lenslet. The sorting method proposed here eliminates such a limitation and extends the dynamic range by tagging each spot in a special sequence.

In vivo fiber-optic confocal reflectance microscope with an injection-molded plastic miniature objective lens.

For in vivo optical diagnostic technologies to be distributed to the developed and developing worlds, optical imaging systems must be constructed of inexpensive components. We present a fiber-optic confocal reflectance microscope with a cost-effective injection-molded plastic miniature objective lens for in vivo imaging of human tissues in near real time. The measured lateral resolution is less than 2.

An array microscope for ultrarapid virtual slide processing and telepathology. Design, fabrication, and validation study.

This paper describes the design and fabrication of a novel array microscope for the first ultrarapid virtual slide processor (DMetrix DX-40 digital slide scanner). The array microscope optics consists of a stack of three 80-element 10 x 8-lenslet arrays, constituting a "lenslet array ensemble." The lenslet array ensemble is positioned over a glass slide.

High resolution, molecular-specific, reflectance imaging in optically dense tissue phantoms with structured-illumination.

Structured-illumination microscopy delivers confocal-imaging capabilities and may be used for optical sectioning in bio-imaging applications. However, previous structured-illumination implementations are not capable of imaging molecular changes within highly scattering, biological samples in reflectance mode. Here, we present two advances which enable successful structured illumination reflectance microscopy to image molecular changes in epithelial tissue phantoms.

Application of the Alvarez-Humphrey concept to the design of a miniaturized scanning microscope.

This paper contains two optical designs that utilize the Alvarez- Humphrey surfaces to provide the miniature multi-modal microscope (4M device) with the additional capability of imaging different object depths onto the same image plane. The Alvarez-Humphrey surfaces are a pair of conjugate, rotationally asymmetrical, aspheric surfaces such that, the lateral movement of these surfaces across the optical axis, results in an element of variable optical power. The first design is a direct application of the Alvarez-Humphrey concept to the 4M device.

Realization of refractive microoptics through grayscale lithographic patterning of photosensitive hybrid glass.

Refractive microlenses with more than 50 microm sag are fabricated using grayscale lithography. Mechanical assembly features are made simultaneously alongside the microlenses to facilitate high precision assembly of miniature optical systems. The microlens elements are formed using lithographic patterning of photosensitive hybrid sol-gel glass requiring no etch transfer to the substrate material.

Endoscopic microscopy.

In vivo endoscopic optical microscopy provides a tool to assess tissue architecture and morphology with contrast and resolution similar to that provided by standard histopathology--without need for physical tissue removal. In this article, we focus on optical imaging technologies that have the potential to dramatically improve the detection, prevention, and therapy of epithelial cancers. Epithelial pre-cancers and cancers are associated with a variety of morphologic, architectural, and molecular changes, which currently can be assessed only through invasive, painful biopsy.

Fiber optic confocal reflectance microscopy: a new real-time technique to view nuclear morphology in cervical squamous epithelium in vivo.

We present a fiber optic confocal reflectance microscope (FCRM) which can be used to image epithelial tissue with sub-cellular resolution in vivo. Confocal images of normal and abnormal appearing cervical tissue were obtained in vivo from eighteen patients undergoing colposcopic examination of the cervix; biopsy specimens were taken from imaged sites. The measured lateral and axial resolutions of the system were 1.

Imaging quality assessment of multi-modal miniature microscope.

We are developing a multi-modal miniature microscope (4M device) to image morphology and cytochemistry in vivo and provide better delineation of tumors. The 4M device is designed to be a complete microscope on a chip, including optical, micro-mechanical, and electronic components. It has advantages such as compact size and capability for microscopic-scale imaging.

Phase grating design for a dual-band snapshot imaging spectrometer.

Infrared spectral features have proved useful in the identification of threat objects. Dual-band focal-plane arrays (FPAs) have been developed in which each pixel consists of superimposed midwave and long-wave photodetectors [Dyer and Tidrow, Conference on Infrared Detectors and Focal Plane Arrays (SPIE, Bellingham, Wash., 1999), pp.

Fiber-optic confocal reflectance microscope with miniature objective for in vivo imaging of human tissues.

We have built a fiber-optic confocal reflectance microscope capable of imaging human tissues in near real time. Miniaturization of the objective lens and the mechanical components for positioning and axially scanning the objective enables the device to be used in inner organs of the human body. The lateral resolution is 2 micrometers and axial resolution is 10 micrometers.

Design of a high-numerical-aperture miniature microscope objective for an endoscopic fiber confocal reflectance microscope.

An endoscopic confocal microscope requires a high-performance, miniaturized microscope objective. We present the design of a miniature water-immersion microscope objective that is approximately 10 times smaller in length than a typical commercial objective. The miniature objective is 7 mm in outer diameter and 21 mm in length (from object to image).