Abstract: A confocal scanning microscope system (10) using cross polarization effects and an enhancement agent (acetic acid) to enhance confocal microscope reflectance images of the nuclei of BCCs (basal cell carcinomas) and SCCs (squamous call carcinomas) in the confocal reflectance images of excised tumor slices. The confocal scanning microscope system having a laser (11) for generating an illumination beam (12), a polygon mirror (18) for scanning the beam to a tissue sample (22) and for receiving a return beam from the tissue sample and detector (28) for detecting the returned beam to form an image. The system further includes a half-waveplate (13) having a rotatable stage (14) and a quarter-wave plate (21) having a rotatable stage (20) disposed in the optical path of the illumination beam and at least a linear polarizer (24) having a rotatable stage (25) disposed in the optical pat of the returned beam from the tissue sample.

Abstract: An imaging apparatus is provided for imaging tissue samples substantially beneath the surface of the tissue sample. The apparatus includes an objective lens and a window defining a tissue contacting surface in pressure contacting relationship with the surface of the tissue sample when the tissue sample is imaged by the objective lens to view tissue structures for pathological applications. The objective lens focuses an illumination beam through the window to the tissue sample and receives returned reflected light of the beam representative of one or more sections of the tissue sample. The apparatus enables a method for in vivo observation of tissue for diagnosis of conditions substantially beneath the surface of the tissue sample. Both two and three-dimensional imaging may be provided for diagnosis and location of basal cell carcinomas and melanomas, and so as to enable visualization of tumor borders prior to excision.

Abstract: A system and method for enhancing images of ex-vivo or in-vivo tissue produced by confocal microscopy, optical coherence tomography, two-photon microscopy, or ultrasound, is provided by applying to the tissue a solution or gel having an effective concentration of citric or other alpha-hydroxy acid which enhances tissue structures, such as cellular nuclei, in such images. Such concentration may be 3–20% acid, and preferably 5% acid.

Abstract: An improved system for confocal imaging within dermal tissue of a patient is provided which minimizes instability in confocal images by reducing the relative motion of the tissue with respect to the confocal imaging optics of the system. The system includes a mechanism for maintaining an area of skin tissue under stress by application of force at the edges of the area, and an imaging head coupled to this mechanism for imaging the stressed skin. The mechanism includes a mechanical structure, such as a platen, brace, or attachment, which both supports the imaging head of the system and applies stress to a limited surface area of the tissue to minimize skin motion during confocal imaging.

Abstract: A tray or holder for tissue specimens, especially of excised tissue, such as biopsied specimens, is used with a confocal imaging system. The tray may be disposable after imaging of the specimen carried therein or may archive the specimen. A window supports the specimen. Clamps mounted inside the tray restrain the tissue. A compliant bag is mounted outside the tray on one side of a window of the tray on which the specimen is disposed. During imaging the specimen is immersed in a liquid contained in the tray having an index of refraction which closely matches the index of refraction of the tissue. The bag also contains an index matching liquid preferably having the same index as the liquid in the tray. Selecting an immersion liquid, which equals the refractive index of the near surface tissue, minimizing wavefront distortion which may result from an effectively corrugated surface of the specimen.

Abstract: A system is provided for marking on a recording medium (36), such as a label, the location of imaged tissue with respect to an exposed surface of the tissue (34). Tissue is imaged by a microscope (11) capable of imaging sections of the tissue below the exposed tissue surface through optics. A ring (32) applied to the surface of the tissue stabilizes the tissue to the optics and localizes a portion of the tissue surface through an aperture in the ring. An actuator (38) is connected to both the ring and the microscope for moving the ring to adjust the position of the tissue with respect to the optics, thereby allowing an operator of the system to survey different images of tissue sections with the microscope.

Abstract: A confocal scanning microscope system (10) using cross polarization effects and an enhancement agent (acetic acid) to enhance confocal microscope reflectance images of the nuclei of BCCs (basal cell carcinomas) and SCCs (squamous call carcinomas) in the confocal reflectance images of excised tumor slices. The confocal scanning microscope system having a laser (11) for generating an illumination beam (12), a polygon mirror (18) for scanning the beam to a tissue sample (22) and for receiving a return beam from the tissue sample and detector (28) for detecting the returned beam to form an image. The system further includes a half-waveplate (13) having a rotatable stage (14) and a quarter-wave plate (21) having a rotatable stage (20) disposed in the optical path of the illumination beam and at least a linear polarizer (24) having a rotatable stage (25) disposed in the optical path of the returned beam from the tissue sample.

Abstract: The quality of images produced by confocal microscopy, and especially scanning laser confocal microscopy, is enhanced especially for images obtained in turbid mediums such as many biological tissue specimens, by reducing speckle from scatterers that exist outside (above and below) the focal plane region which is being imaged by utilizing sheared beams, both of which are focused to spots in the focal or image plane (region of interest) and polarizing the beams to have opposite senses of circular polarization (right and left handed circular polarization). The return light from the image plane of certain polarization is detected after passing through the confocal aperture of the confocal microscope. Light from scatterers outside the region of interest, which are illuminated by both of the sheared beams, interfere thereby reducing speckle due to such scatterers, and particularly scatters which are adjacent to the image plane.

Abstract: In order to facilitate pathological examination of a lesion in in-vivo tissue, a system and method are provided having a computer system in which both a camera for producing a digital macroscopic picture of the lesion and an imager are coupled to the computer system. The imager is responsive to the computer system and has optics for scanning the lesion to generate images representing microscopic sections of the lesion which provide sufficient information for pathological examination of the lesion. The computer system generates location information, referencing the location in the macroscopic picture of the lesion where the lesion was scanned to the images, and stores data in an electronic file structure which contains at least a representation of the images, a representation of the macroscopic picture, and the location information.

Abstract: An improved system for cellular surgery which includes a laser for producing a laser beam, and confocal optics for scanning and focusing the laser beam in tissue and generating confocal images of the tissue in accordance with returned light from the tissue. The confocal images are visualized on a display. The system includes a controller for enabling the operator to select one or more cells of the tissue in the displayed confocal images for surgical treatment. The controller operates the laser and confocal optics in a first mode to treat the tissue when the confocal optics focus the laser beam at least one region associated with the selected cells in the tissue, but at all other times operates the laser and confocal optics in a second mode which does not damage the tissue.

Abstract: The quality of images produced by confocal microscopy, and especially scanning laser confocal microscopy, is enhanced especially for images obtained in turbid mediums such as many biological tissue specimens, by reducing speckle from scatterers that exist outside (above and below) the section which is being imaged by utilizing sheared beams, both of which are focused to laterally or vertically offset spots and polarizing the beams to have opposite senses of circular polarization (right and left handed circular polarization). The return light from the section of certain polarization is detected after passing through the confocal aperture of the confocal microscope. Images can be formed using optical coherence detection of the return light. Light from scatterers outside the section of interest, which are illuminated by both of the sheared beams, interfere thereby reducing speckle due to such scatterers, and particularly scatters which are adjacent to the section being imaged.

Abstract: An improved system for confocal imaging within dermal tissue of a patient is provided which minimizes instability in confocal images by reducing the relative motion of the tissue with respect to the confocal imaging optics of the system. The system includes a mechanism for maintaining an area of skin tissue under stress by application of force at the edges of the area, and an imaging head coupled to this mechanism for imaging the stressed skin. The mechanism includes a mechanical structure, such as a platen, brace, or attachment, which both supports the imaging head of the system and applies stress to a limited surface area of the tissue to minimize skin motion during confocal imaging.

Abstract: A confocal microscope is provided including a confocal imaging system having an objective lens, and a device which can be an attachment to the confocal imaging system, or a part thereof. The device has a housing with an interior cavity and two opposite ends, where one end of the housing receives the objective lens, and at the other end of the housing which is attached to, or is a part of, the housing which forms that other end. The chamber has first and second plates presenting two opposing sides of the chamber, where the first plate lies adjacent the tissue to be treated, and the second plate is attached to the housing. The interior cavity of the housing has a window defining a first volume between the window and the objective lens, and a second volume between the window and the chamber. The first volume has an immersion liquid optically matched to the objective lens, and the second volume is approximately evacuated.

Abstract: A cassette for retaining a specimen of surgically exposed tissue from a patient in an orientation that facilitates optical sectioning of the tissue by a confocal microscopic or other optical imaging microscope. The cassette includes a base member having a rigid optically transparent window upon which a tissue specimen is situated, a pliable membrane locatable over a substantial portion of the base member including the window, and an upper member, having an aperture therethrough, which can cover the base member to provide an enclosed cavity between the membrane and the window sealing the tissue specimen therein. With the tissue specimen in the enclosed cavity of the cassette, the edges of the tissue specimen may be positioned planar against the window and retained in that position by bonds formed between the membrane and window at multiple points or locations around the tissue specimen.

Abstract: A system is provided having a clamping apparatus with first and second members, which are spaced apart from each other. An attachment mechanism is connected to the first member for attaching the apparatus to a confocal imaging system. This attachment mechanism has a surface with a window facing the tissue to be examined. Opposing this surface is a third member connected to the second member. A clamping mechanism includes the first and second members for clamping the tissue between the surface of the attachment mechanism and the surface of the third member, thereby stabilizing the tissue to the confocal imaging system. The window of the attachment mechanism presents the clamped tissue to the confocal imaging system.

Abstract: A tray or holder for tissue specimens, especially of excised tissue, such as biopsied specimens, is used with a confocal imaging system, especially a laser scanning confocal microscope system. The tray may be disposable after imaging of the specimen carried therein or may archive the specimen. A window supports the specimen. Clamps mounted inside the tray restrain the tissue. A compliant bag is mounted outside the tray on one side of a window of the tray on which the specimen is disposed. During imaging the specimen is immersed in a liquid contained in the tray having an index of refraction which closely matches the index of refraction of the tissue. The bag also contains an index matching liquid preferably having the same index as the liquid in the tray. A stabilizing plate is attached to a surface of the bag which faces the window.

Abstract: The quality of image produced by confocal microscopy, and especially scanning laser confocal microscopy, is enhanced especially for images obtained in turbid mediums such as many biological tissues specimens, by reducing speckle from scatterers that exist outside (above and below) the section which is being imaged by utilizing reduced coherence illumination, such as provided by a multi-mode laser. The laser beam is focused to provide its intensity in lobes forming offset spots in opposite (180°) amplitude phase relationship. The lobes are combined in the return light from the section and detected after passing through the confocal aperture of the confocal microscope. Images can be formed from the detected return light. Light from scatterers outside the section of interest, which are illuminated by both of the lobes beams overlap outside the section and interfere, thereby reducing speckle due to such scatterers, and particularly scatters which are adjacent to the section being imaged.

Abstract: A handheld imaging microscope for imaging tissue samples substantially beneath the surface of the tissue sample. The microscope includes an objective lens and a window defining a tissue contacting surface in pressure contacting relationship with the surface of the tissue sample when the tissue sample is imaged by the objective lens to view tissue structures for pathological applications. The objective lens focuses an illumination beam through the window to the tissue sample and receives returned reflected light of the beam representative of one or more sections of the tissue sample.