US20070070497A1 - System and method for enhancing confocal reflectance images of tissue specimens - Google Patents

System and method for enhancing confocal reflectance images of tissue specimens Download PDF

Info

Publication number
US20070070497A1
US20070070497A1 US11/601,203 US60120306A US2007070497A1 US 20070070497 A1 US20070070497 A1 US 20070070497A1 US 60120306 A US60120306 A US 60120306A US 2007070497 A1 US2007070497 A1 US 2007070497A1
Authority
US
United States
Prior art keywords
tissue
confocal
method according
light
solution
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
US11/601,203
Inventor
Jay Eastman
Milind Rajadhyaksha
James Zavislan
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Zavislan James M
Eastman Jay M
Original Assignee
Eastman Jay M
Milind Rajadhyaksha
Zavislan James M
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority to US24114000P priority Critical
Priority to US09/978,957 priority patent/US7139122B1/en
Application filed by Eastman Jay M, Milind Rajadhyaksha, Zavislan James M filed Critical Eastman Jay M
Priority to US11/601,203 priority patent/US20070070497A1/en
Publication of US20070070497A1 publication Critical patent/US20070070497A1/en
Application status is Abandoned legal-status Critical

Links

Images

Classifications

    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS, OR APPARATUS
    • G02B21/00Microscopes
    • G02B21/0004Microscopes specially adapted for specific applications
    • G02B21/002Scanning microscopes
    • G02B21/0024Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0052Optical details of the image generation
    • G02B21/0068Optical details of the image generation arrangements using polarisation
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K49/00Preparations for testing in vivo
    • A61K49/001Preparation for luminescence or biological staining
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57407Specifically defined cancers
    • G01N33/5743Specifically defined cancers of skin, e.g. melanoma

Abstract

A system using cross polarization effects and an enhancement agent having citric or other similar alpha hydroxy acid to enhance confocal microscope reflectance images and particularly images of the nuclei of BCCs (basal cell carcinomas) and SCCs (squamous cell carcinomas) in the confocal reflectance images of excised tumor slices obtained during Mohs surgery by illuminating the tissue being imaged (a tumor slice) using polarized light. The reflected illumination is passed to a polarization analyzer, which passes the polarization component which is crossed with respect to the polarization of the illuminating light. The light from the analyzer is passed through the confocal aperture and detected. The section of the tissue either at the surface or within the tissue is scanned and the reflectance image is produced with enhanced visualization of the cellular or nuclear structure thereof thereby enabling determination of the extent of the tumor (cancerous cells) in the section. A method is also provided using the system for diagnosing cancerous cells in skin tissue.

Description

  • This application claims the benefit of priority to U.S. Provisional Patent Application No. 60/241,140, filed Oct. 17, 2000, which is herein incorporated by reference, and describes an improvement over International Application No. PCT/US00/07008, which was filed on Sep. 14, 2001 in the United States Patent and Trademark Office as U.S. patent application Ser. No. 09/936,535.
  • FIELD OF THE INVENTION
  • The present invention relates to confocal microscopy and particularly to a system (method and apparatus) for enhancing images of tissue at the surface or internally of a tissue sample so as to enable rapid and accurate screening of tissue for the determination of the nuclear and cellular structure thereof. The present invention also relates to a method for diagnosing cancerous cells in skin tissue using confocal microscopy. The invention is especially suitable in providing enhanced images of the nuclei of BCC/SCC (basal cell carcinoma or squamous cell carcinoma) in confocal reflectance images of tumor slices obtained during Mohs micrographic surgery. Tissue may be either naturally exposed, or surgically excised tissue.
  • BACKGROUND OF THE INVENTION
  • Mohs micrographic surgery for BCCs and SCCs involves precise excision of the cancer with minimal damage to the surrounding normal skin. Conventionally, precise excision is guided by histopathologic examination for cancer margins in the excised tissue slices during Mohs surgery. Typically, 2-4 slices are excised, and there is a waiting time of 10-30 minutes for the surgeon and patient while each slice is being processed.
  • Confocal reflectance microscopes can noninvasively image nuclear and cellular detail in living skin to provide images of tissue sections, such a microscope is described in U.S. Pat. No. 5,880,880. The contrast in the images is believed to be due to the detected variations in the singly back-scattered light caused by variations in refractive indices of tissue microstructure. Within the epidermal (basal and squamous) cells, the cytoplasm appears bright and the nuclei as dark ovals. The underlying dermis consists of collagen bundles and that, too, appears bright with dark spaces in-between. Thus, when neoplastic epidermal cells invade the dermis as in BCCs and SCCs, confocal detection of the cancers is very difficult because the cells and nuclei lack contrast relative to the surrounding normal dermis.
  • Acetic acid (vinegar) has been used as an image enhancement agent which enhances the back-scatter of light from certain cells. This is described, for example, in U.S. Pat. No. 5,733,739. When a tissue is illuminated with light of one polarization and a reflected image is detected via light of cross polarization the contrast of certain cells in the detected image is enhanced, when the tissue is treated by acetic acid, especially when the image is viewed via cross-polarized light in a confocal microscope. Such a system is described in the above referenced International Application No. PCT/US00/07008.
  • SUMMARY OF THE INVENTION
  • It is the feature of the present invention to provide an improved system and method for confocal microscopy by cross polarizing the light illuminating a tissue sample and the light returned from the tissue sample representing a section of the tissue and using an image enhancement agent other than acetic acid.
  • It is another feature of the present invention to use such cross polarizing of the light illuminating a tissue sample and the light returned from the tissue sample in combination with imaging the sample when immersed in an image enhancement agent containing citric acid or another alpha hydroxy acid.
  • It is a further feature of the present invention to provide a method for diagnosing cancerous cells in skin tissue using confocal microscopy and a citric or alpha hydroxy acid image enhancing agent to treat the tissue.
  • Briefly described, a system for providing enhanced images in confocal microscopy is provided by utilizing cross polarized light in the illumination of tissue and in the detection of light from which the images are formed, respectively, and where an image enhancing agent, which contains an effective amount of citric or similar alpha hydroxy acid, is used in a bath in which the tissue is immersed while being imaged. It will be understood that the agent may also be applied to tissue by coating, dipping or encapsulating in a gel of citric or other alpha hydroxy acid solution.
  • It has been found in accordance with the invention that a confocal laser scanning microscope using cross polarized components of light in illumination and in the detection of the reflected light from tissue specimens immersed in such an enhancement agent solution images of the cellular structure are enhanced, enabling cells and voids in the structure and the cell condition to be readily observed. By virtue of the use of such cross polarized light in imaging of tumor slices obtained in the course of Mohs surgery, epidermis sections which may have holes in the collagen are imaged more accurately so that holes are unlikely to be confused with cells or cell structure.
  • A method is also provided for detecting cancerous basal cell and squamous cell in dermal tissue with confocal reflected light imaging having the steps of: washing the tissue to be imaged with a solution of citric or other alpha hydroxy acid to whiten epithelial cells and believe to compact the chromatin of the tissue; imaging the tissue with a confocal microscope to provide confocal images of basal and squamous cells in which the confocal microscope directs light into the tissue and collects reflected light representing confocal images of the tissue; changing the polarization state of the light used by the confocal microscope to increase the contrast of the nuclei of basal and squamous cells in the confocal images; and analyzing the nuclei of the basal and squamous cells in the confocal images to diagnose which of such cells are cancerous.
  • BRIEF DESCRIPTION OF THE DRAWINGS
  • The foregoing features and advantages of the invention will become more apparent from a reading of the following description in connection with the accompanying drawings wherein,
  • FIG. 1 is a schematic diagram of a Vivascope (TM) confocal microscope which is available from Lucid Inc. of Rochester, N.Y. and is described in the above referenced U.S. Pat. No. 5,880,880; and
  • FIGS. 2A, B, C, and D are schematic depictions of various parts of the confocal microscope system and the cross-polarized illumination which is used therein.
  • Detailed Description of Invention
  • Referring to the drawings, in the confocal microscope 10 of FIG. 1, a linearly polarized (p-state) laser beam 12 is passed through a half wave plate (HWP) 13 on a rotation stage 14. A confocal microscope especially suitable in practicing the invention is described in U.S. Pat. No. 5,880,880, issued Mar. 9, 1999, which is herein incorporated by reference. Other confocal microscopes may also be used. The illumination through the non-polarizing or partially polarizing beam splitter 16 is scanned, as by a polygon mirror 18 and galvanometric mirror 19 across the specimen or sample 22 having a surface 22 a. As shown in FIGS. 2B and 2C, sample 22 may be a BCC/SCC sample in a sample holder or container 22 b contained in an enhancement solution bath 26 having water 28 under a tissue ring 33 which places the sample 22 under tension. As shown in FIG. 1, the microscope 10, via an objective lens 23, images the tissue sample 23 through an opening 33 a in the tissue ring 33. For example, the opening 33 a may include a window having a material transparent to the beam.
  • The target surface is the surface of the sample 22 (such as a tissue tumor specimen), which may be at the surface 22 a or within the body of the sample, utilizing the techniques described in the above referenced U.S. patent. The polarization of the incident light and the reflected light also can be modified using a quarter wavelength plate (QWP) 21 which is also removably mounted on a rotation stage 20.
  • The detected light is cross-polarized that is in the s-state as shown by the bulls-eye indication 12 a in FIG. 1 and labeled “detection s-state” in FIG. 2D. It is crossed or perpendicular or orthogonal to the p-state. Although preferably cross-polarized light is in s and p states, because the beam splitter may be non-polarizing or partially polarizing, other states are possible. The detected illumination of desired polarization is obtained with an analyzer 24 also mounted on a rotation stage 25. For example, analyzer 24 may be a linear polarizer. The light from the analyzer 24 is passed through the confocal aperture 28 a, such as a pinhole, and a photo-detector 28, such as an avalanche photodiode (APD) in FIG. 1. While p polarized light from a linearly polarized laser 11 is shown in FIG. 1, the linearly polarized laser 11 and the half wave plate 13 can be replaced with a laser providing an unpolarized laser beam and a linear polarizer, respectively. Further, the linear polarizer and the analyzer 24 can then be replaced with a polarized beam splitter. Also, instead of rotating the half wave plate 13 and the analyzer 24, they can be kept fixed in cross polarization states and the sample 22 can be rotated.
  • As shown in FIG. 1, optical components are provided in confocal microscope 10 to direct the beam from laser 11 along a path to sample 22, and include, beam expander-spatial filter 42 (which, for example, may be provided by two lens 42 a and 42 b and aperture 42 c), HWP 13, mirror 43, ND filter 44 (which, for example, may be a neutral density filter, such as provided by a circular variable attenuator manufactured by Newport Research Corporation), through beam splitter 16 to polygon mirror 18. The beam is then deflected by polygon mirror 18 through a lens 45 (which for example, may be a f/2 lens), a lens 46 (which for example, may be a f/5.3 lens), and deflected by galvanometric mirror 19 through a lens 47 (which for example, may be a f/3 lens), QWP 21 and objective lens 23 to sample 22. The optical components along the path of the reflected light returned from the sample 22 to detector 28 include, objective lens 23, QWP 21, lens 47, and deflected by mirrors 19 and 18 via lenses 46 and 45 to beam splitter 16. The beam splitter 16 directs the returned light through lens 48, analyzer 24, and pinhole 28 a to detector 28. The raster line 17 a and raster plane 17 b in FIG. 1 are illustrated by dashed lines to denote the angular scan of the beam along a raster line 17 a generated by the rotation of polygon mirror 18, while the angular movement of galvanometric mirror 19 scans that raster line to form a raster plane 17 b. In this manner, a confocal image of a tissue section can be captured by the control electronics 38 through detector 28. To provide a start of scan beam 12 c to synchronize the control electronics 38 with the start of each raster line, the beam splitter 16 directs part of the beam incident the beam splitter 16 to rotating polygon mirror 18, via mirror 48, to split diode 50 (e.g., photo-diode) which is connected to the control electronics 38 to provide a start of scan pulse at the beginning of each raster line. Motors, not shown, can provide the desired rotation and angular movement of respective mirrors 18 and 19.
  • The system which is shown in FIG. 1 operates as follows:
  • 1. Remove QWP 21. Rotate the HWP 13 so that its fast axis is at 90 degrees with the illumination p-state (see FIG. 2A). Thus, there is no change (rotation) of the direction of the p-state. Rotate the analyzer 24 so that it acts as a crossed polarizer and transmits the detection s-state (which is orthogonal to the illumination p-state).
  • 2. The surgically excised tissue sample 22 is placed in a water bath 26 with a tissue-ring 33 placed on top (see FIG. 2B).
  • 3. The water bath 26 containing the sample 22 is placed under the objective lens 23, such that the tissue-ring 33 fits into the objective lens housing 31 (see FIG. 2C). The water bath 26 is on an XY translation stage 34 to move the sample 22. The XY stage 34 is on a lab-jack 35 with which can move the entire assembly 36 upwards, such that the sample 22 is gently pressed between the tissue-ring 33 and the water bath 26 to keep the sample 22 still during the imaging. Arrow 37 denotes the direction of such light pressure.
  • 4. Rotate the HWP 13 in small angular increments of 10 degrees and, correspondingly, the analyzer 24 in angular increments of 20 degrees, on their respective stages 14 and 25, such that the analyzer 24 is always cross-polarized with respect to the illumination polarization state. The confocal images of the sample 22 change from bright to dark to bright as the HWP 13 and analyzer 24 is rotated.
  • 5. Set the HWP 13 and analyzer 24 such that the sample 22 appears dark (i.e., minimum brightness). Survey the sample 22 by moving it with the XY stage 34, to check that the sample appears dark everywhere in the confocal images.
  • 6. Lower the water bath 26 using the lab-jack 35. Remove the water from within the tissue ring 33, and add an enhancement agent, namely citric or other alpha hydroxy acid (e.g., to provide a 5% by volume—ph 2.5—solution in the water). Raise the lab-jack 35 and place the sample 22, as before, under the objective lens 23. It will be understood that the concentration of the solution to be effective may be in the range of from 3-20%.
  • 7. Survey the sample 22 by moving it with the XY stage 34, and focusing on the surface and at varying depths of the sample with the objective lens 23 (which may be mounted on a Z-translation stage to move the objective lens towards and away from the sample). Confocal images are either videotaped or grabbed in this “crossed polarization” mode at a frame grabber 39, video monitor 40, or videotape recorder 41 via control electronics 38.
  • 8. Whenever or wherever necessary, confocal images are obtained in “brightfield” mode, to either determine lateral or depth location, or identify structures (examples: hair follicles, sweat ducts, epithelial margins) within the sample. (This is analogous to using reflectance imaging in conjunction with fluorescence imaging.) The QWP 21 is inserted and rotated so that its optic axis is at 45 degrees to both the illumination and detection linear polarization states (see FIG. 2D).
  • With the confocal reflectance light microscope 10 described herein, BCCs, SCCs in human skin are described herein without the processing (fixing, sectioning, staining) that is required for conventional histopathology of Mohs surgery. Rapid confocal detection is provided after strongly enhancing the contrast of nuclei in the cancer cells relative to the surrounding normal tissue using citric or other alpha hydroxy acid and crossed polarization.
  • To improve the detection of BCCs and SCCs in confocal images in tissue, such as dermal tissue, which may be either naturally exposed, or surgically excised, the contrast of the nuclei of such cells is increased by the following method. The area of the tissue to be imaged is washed with 5% citric or alpha hydroxy acid, as described earlier. Citric or alpha hydroxy acid causes whitening of epithelial tissue and believe to cause the compaction of the chromatin. The chromatin-compaction is believed to increase its refractive index, which then increases light back-scatter from the nuclei and makes them appear bright. Next, the tissue area is imaged with confocal microscope 10 in which the polarization state of the light directed to the tissue and collected by the confocal microscope is controlled by rotating the linear polarizer of analyzer 24. When illuminated with linearly polarized light and confocally imaged through the analyzer 24, the brightness of the citric or alpha hydroxy acid-stained nuclei does not vary much, whereas the brightness of the collagen varies from maximum to minimum. The back-scattered light from the inter-nuclear structure is significantly depolarized (probably due to multiple scattering), whereas that from the dermis preserves the illumination polarization (due to single back-scatter). With the light in a crossed polarized state, bright nuclei in the BCCs and SCCs are shown in the confocal images produced by the microscope in strong contrast against a dark background of surrounding normal dermis. BCCs and SCCs can be distinguished from normal tissue by the cellular organization, cell size, cell shape, nuclear morphology, and cellular differentiation. One example of cellular organization is anaplasia. One example of cell size and shape and nuclear morphology is dysplasia. One example of cellular differentiation is pleomorphism.
  • Thus, the bright clusters of nuclei in the cancer cells are detectable at low resolution, as in conventional histopathology. Mosaics of low-resolution confocal images can be assembled to produce confocal maps of the BCCs or SCCs within the entire excised tissue. Detection of the cancers is made within minutes; thus, the total savings in time for a Mohs surgery can be hours.
  • Others cancers and tissue abnormalities may also be detected by using this approach any time a cellular tissue needs to be distinguished from acellular background. For example, dermal melanocytes, mucosal tissue in stromal tissue, breast epithelium in a stromal matrix.
  • From the foregoing description, it will be apparent that an improved system for enhancing images in confocal microscopy and method for diagnosing skin cancer cells is provided using citric or other alpha hydroxy acid. Variations and modifications in the herein described system and method will undoubtedly become apparent to those skilled in the art. Accordingly, the foregoing description should be taken as illustrative and not in a limiting sense.

Claims (15)

1-7. (canceled)
8. A method for confocal imaging tissue comprising the steps of:
generating an illumination beam;
scanning the beam to the tissue;
receiving returned light from the tissue representing of a section of the tissue;
cross polarizing the illumination beam and the returned light with respect to each other;
detecting the returned light to form an image of the section of the tissue, and the step of treating said sample with a solution of alpha hydroxy acid which enhances the brightness of one or more tissue structures in the image of the section of the tissue.
9. The method according to claim 8 wherein said polarization controlling step further comprises the step of changing polarization state of at least one o the illumination beam and the returned light to effect characteristics of tissue structures in the image of the tissue section to enable determination of which of the tissue structures are cancerous.
10. The method according to claim 9 wherein said tissue is skin tissue.
11. The method according to claim 8 wherein the tissue is one of naturally exposed tissue and surgically excised tissue.
12. The method according to claim 8 further comprising the step of applying said solution to said tissue.
13. The method according to claim 12 wherein said solution is applied to said tissue by one of coating, washing, dipping, or rinsing with said image enhancing agent.
14. The method according to claim 12 wherein said solution is provided by a gel encapsulating said tissue.
15. The method according to claim 12 wherein said solution is applied by placing said tissue in a bath of said solution.
16. The method according to claim 8 wherein said solution is of 3-20% concentration of said acid.
17. The method according to claim 8 wherein said solution is of 5% concentration of said acid.
18. The method according to claim 8 further comprising the step of analyzing cells in said images to diagnose which of said cells are cancerous.
19. A method for detecting cancerous basal cell and squamous cell in dermal tissue with confocal reflected light imaging, said method comprising the steps of:
washing the tissue to be imaged with a solution of citric or other alpha hydroxy acid which whitens epithelial cells and compacts cromatin of the tissue;
imaging the tissue with a confocal microscope to provide confocal images of basal and squamous cells in which the confocal microscope directs light into the tissue and collects reflected light representing confocal images of the tissue;
changing the polarization state of the light used by the confocal microscope to increase the contrast of the nuclei of basal and squamous cells in the confocal images; and
analyzing the nuclei of the basal and squamous cells in the confocal images to diagnose which of such cells are cancerous.
20-36. (canceled)
37. The method according to claim 8 wherein said alpha hydroxy acid represents citric acid.
US11/601,203 2000-10-17 2006-11-17 System and method for enhancing confocal reflectance images of tissue specimens Abandoned US20070070497A1 (en)

Priority Applications (3)

Application Number Priority Date Filing Date Title
US24114000P true 2000-10-17 2000-10-17
US09/978,957 US7139122B1 (en) 2000-10-17 2001-10-16 System and method for enhancing confocal reflectance images of tissue specimens
US11/601,203 US20070070497A1 (en) 2000-10-17 2006-11-17 System and method for enhancing confocal reflectance images of tissue specimens

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
US11/601,203 US20070070497A1 (en) 2000-10-17 2006-11-17 System and method for enhancing confocal reflectance images of tissue specimens

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
US09/978,957 Division US7139122B1 (en) 2000-10-17 2001-10-16 System and method for enhancing confocal reflectance images of tissue specimens

Publications (1)

Publication Number Publication Date
US20070070497A1 true US20070070497A1 (en) 2007-03-29

Family

ID=37423250

Family Applications (2)

Application Number Title Priority Date Filing Date
US09/978,957 Active 2023-01-30 US7139122B1 (en) 2000-10-17 2001-10-16 System and method for enhancing confocal reflectance images of tissue specimens
US11/601,203 Abandoned US20070070497A1 (en) 2000-10-17 2006-11-17 System and method for enhancing confocal reflectance images of tissue specimens

Family Applications Before (1)

Application Number Title Priority Date Filing Date
US09/978,957 Active 2023-01-30 US7139122B1 (en) 2000-10-17 2001-10-16 System and method for enhancing confocal reflectance images of tissue specimens

Country Status (1)

Country Link
US (2) US7139122B1 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101915994A (en) * 2010-07-15 2010-12-15 苏州巨像科技有限公司 Method for enhancing brightness of laser scanning projected image

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7003345B1 (en) * 2000-10-17 2006-02-21 Lucid, Inc. System and method for enhancing microscope images of tissue using citric acid and agents of the like
US20040133112A1 (en) * 2002-03-08 2004-07-08 Milind Rajadhyaksha System and method for macroscopic and confocal imaging of tissue
US8260401B2 (en) 2006-07-26 2012-09-04 University Of Rochester Non-invasive in-vivo imaging of mechanoreceptors in skin using confocal microscopy
US9677869B2 (en) 2012-12-05 2017-06-13 Perimeter Medical Imaging, Inc. System and method for generating a wide-field OCT image of a portion of a sample

Citations (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4298590A (en) * 1975-02-25 1981-11-03 Samuel Bogoch Detection of malignant tumor cells
US4348679A (en) * 1980-10-06 1982-09-07 United Technologies Corporation Multi-mode dual-feed array radar antenna
US4647447A (en) * 1981-07-24 1987-03-03 Schering Aktiengesellschaft Diagnostic media
US4863226A (en) * 1987-03-13 1989-09-05 Nederlandse Organisatie Voor Toegepas - Natuurwetenschappelijk Onderzoek Tno Confocal laser scanning microscope
US5260569A (en) * 1991-07-25 1993-11-09 Fuji Photo Film Co., Ltd. Scanning microscope and scanning mechanism
US5496535A (en) * 1991-04-12 1996-03-05 Alliance Pharmaceutical Corp. Fluorocarbon contrast media for use with MRI and radiographic imaging
US5733739A (en) * 1995-06-07 1998-03-31 Inphocyte, Inc. System and method for diagnosis of disease by infrared analysis of human tissues and cells
US5880880A (en) * 1995-01-13 1999-03-09 The General Hospital Corp. Three-dimensional scanning confocal laser microscope
US5995867A (en) * 1997-03-19 1999-11-30 Lucid Inc Cellular surgery utilizing confocal microscopy
US6134009A (en) * 1997-11-07 2000-10-17 Lucid, Inc. Imaging system using polarization effects to enhance image quality
US6134010A (en) * 1997-11-07 2000-10-17 Lucid, Inc. Imaging system using polarization effects to enhance image quality
US6187289B1 (en) * 1997-10-20 2001-02-13 Board Of Regents, The University Of Texas System Acetic acid as a contrast in reflectance confocal imaging of tissue
US6264914B1 (en) * 1996-10-28 2001-07-24 Nycomed Imaging As Contrast agents
US6319488B1 (en) * 1995-10-11 2001-11-20 Institut für Diagnostikforschung GmbH an der Freien Universität Berlin Contrast medium for near infrared diagnosis
US6348325B1 (en) * 1999-10-29 2002-02-19 Cytyc Corporation Cytological stain composition
US6498945B1 (en) * 1997-05-19 2002-12-24 Amersham Health As Sonodynamic therapy using an ultrasound sensitizer compound
US6720547B1 (en) * 1999-03-18 2004-04-13 Lucid, Inc. System and method for enhancing confocal reflectance images of tissue specimens
US7003345B1 (en) * 2000-10-17 2006-02-21 Lucid, Inc. System and method for enhancing microscope images of tissue using citric acid and agents of the like

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3422144A1 (en) 1984-06-14 1985-12-19 Bille Josef Geraet for presentation flaechenhafter areas of the human eye
US5788639A (en) 1995-07-13 1998-08-04 Lucid Technologies, Inc. Confocal imaging through thick dermal tissue
CN1072019C (en) 1997-03-11 2001-10-03 丁炜 Contrast medium capable of eliminating in-stomach fluid level false shadow in CT scanning
AU4012500A (en) 1999-03-18 2000-10-04 Lucid, Inc. System and method for enhancing confocal reflectance images of tissue specimens

Patent Citations (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4298590A (en) * 1975-02-25 1981-11-03 Samuel Bogoch Detection of malignant tumor cells
US4348679A (en) * 1980-10-06 1982-09-07 United Technologies Corporation Multi-mode dual-feed array radar antenna
US4647447A (en) * 1981-07-24 1987-03-03 Schering Aktiengesellschaft Diagnostic media
US4863226A (en) * 1987-03-13 1989-09-05 Nederlandse Organisatie Voor Toegepas - Natuurwetenschappelijk Onderzoek Tno Confocal laser scanning microscope
US5496535A (en) * 1991-04-12 1996-03-05 Alliance Pharmaceutical Corp. Fluorocarbon contrast media for use with MRI and radiographic imaging
US5260569A (en) * 1991-07-25 1993-11-09 Fuji Photo Film Co., Ltd. Scanning microscope and scanning mechanism
US5880880A (en) * 1995-01-13 1999-03-09 The General Hospital Corp. Three-dimensional scanning confocal laser microscope
US5733739A (en) * 1995-06-07 1998-03-31 Inphocyte, Inc. System and method for diagnosis of disease by infrared analysis of human tissues and cells
US6319488B1 (en) * 1995-10-11 2001-11-20 Institut für Diagnostikforschung GmbH an der Freien Universität Berlin Contrast medium for near infrared diagnosis
US6264914B1 (en) * 1996-10-28 2001-07-24 Nycomed Imaging As Contrast agents
US5995867A (en) * 1997-03-19 1999-11-30 Lucid Inc Cellular surgery utilizing confocal microscopy
US6498945B1 (en) * 1997-05-19 2002-12-24 Amersham Health As Sonodynamic therapy using an ultrasound sensitizer compound
US6187289B1 (en) * 1997-10-20 2001-02-13 Board Of Regents, The University Of Texas System Acetic acid as a contrast in reflectance confocal imaging of tissue
US6241662B1 (en) * 1997-10-20 2001-06-05 Lifespex, Inc. Acetic acid as a signal enhancing contrast agent in fluorescence spectroscopy
US6134009A (en) * 1997-11-07 2000-10-17 Lucid, Inc. Imaging system using polarization effects to enhance image quality
US6134010A (en) * 1997-11-07 2000-10-17 Lucid, Inc. Imaging system using polarization effects to enhance image quality
US6720547B1 (en) * 1999-03-18 2004-04-13 Lucid, Inc. System and method for enhancing confocal reflectance images of tissue specimens
US6348325B1 (en) * 1999-10-29 2002-02-19 Cytyc Corporation Cytological stain composition
US7003345B1 (en) * 2000-10-17 2006-02-21 Lucid, Inc. System and method for enhancing microscope images of tissue using citric acid and agents of the like

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101915994A (en) * 2010-07-15 2010-12-15 苏州巨像科技有限公司 Method for enhancing brightness of laser scanning projected image

Also Published As

Publication number Publication date
US7139122B1 (en) 2006-11-21

Similar Documents

Publication Publication Date Title
US3517980A (en) Method and arrangement for improving the resolving power and contrast
Salomatina et al. Optical properties of normal and cancerous human skin in the visible and near-infrared spectral range
US5014709A (en) Method and apparatus for high resolution holographic imaging of biological tissue
AU732108B2 (en) Video imaging of superficial biological tissue layers using polarized light
Cicchi et al. Contrast and depth enhancement in two-photon microscopy of human skin ex vivo by use of optical clearing agents
US7725169B2 (en) Contrast enhanced spectroscopic optical coherence tomography
Lichtman Confocal microscopy
Pan et al. Non-invasive imaging of living human skin with dual-wavelength optical coherence tomography in two and three dimensions
US20110115895A1 (en) Multidirectional selective plane illumination microscopy
US6609015B2 (en) Analysis of a composition
US8454512B2 (en) Confocal photoacoustic microscopy with optical lateral resolution
Strupler et al. Second harmonic imaging and scoring of collagen in fibrotic tissues
US5305759A (en) Examined body interior information observing apparatus by using photo-pulses controlling gains for depths
JP6046325B2 (en) Apparatus for the method and method for progressively in one or more biological samples by increasing the resolution observation and analysis
US6215587B1 (en) Microscope imaging inside highly scattering media
US7264794B2 (en) Methods of in vivo cytometry
US20050240107A1 (en) Detecting human cancer through spectral optical imaging using key water absorption wavelengths
US7218446B2 (en) Imaging system having a fine focus
Liang et al. Fiber confocal reflectance microscope (FCRM) for in-vivo imaging
Amos et al. Use of confocal imaging in the study of biological structures
US7460248B2 (en) Tissue imaging system
AU752872B2 (en) Confocal imaging through thick dermal tissue
Corcuff et al. In vivo vision of the human skin with the tandem scanning microscope
González et al. Real-time, in vivo confocal reflectance microscopy of basal cell carcinoma
König Clinical multiphoton tomography

Legal Events

Date Code Title Description
STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION