Color translating UV microscope

Abstract

A color translating UV microscope for research and clinical applications involving imaging of living or dynamic samples in real time and providing several novel techniques for image creation, optical sectioning, dynamic motion tracking and contrast enhancement comprises a light source emitting UV light, and visible and IR light if desired. This light is directed to the condenser via a means of selecting monochromatic, bandpass, shortpass, longpass or notch limited light. The condenser can be a brightfield, darkfield, phase contrast or DIC. The slide is mounted in a stage capable of high speed movements in the X, Y and Z dimensions. The microscope uses broadband, narrowband or monochromat optimized objectives to direct the image of the sample to an image intensifier or UV sensitive video system. When an image intensifier is used it is either followed by a video camera, or in the simple version, by a synchronized set of filters which translate the image to a color image and deliver it to an eyepiece for viewing by the microscopist. Between the objective and the image intensifier there can be a selection of static or dynamic switchable filters. The video camera, if used, produces an image which is digitized by an image capture board in a computer. The image is then reassembled by an overlay process called color translation and the computer uses a combination of feedback from the information in the image and operator control to perform various tasks such as optical sectioning and three dimensional reconstruction, coordination of the monochromater while collecting multiple images sets called image planes, tracking dynamic sample elements in three space, control of the environment of the slide including electric, magnetic, acoustic, temperature, pressure and light levels, color filters and optics, control for microscope mode switching between transmitted, reflected, fluorescent, Raman, scanning, confocal, area limited, autofluorescent, acousto-optical and other modes.

1 . A microscope for translating spectral information to a visible color image in which light from a source is separated into components by either a set of two or more filters or a device for providing wavelength limited light and then passed through or reflected off an sample then imaged by an objective lens onto a video camera where it is converted to visible light by a fluorescent coating on the photosensitive surface of video camera which provides the image as an electronic signal which is then converted into electronic data by a video to computer interface system and then recombined into a multicolor image by computer processing finally creating a color visible image on a display monitor where the computer is supplied with information on the position of the filters or wavelength limited light in order to synchronize acquisition of the images and the color translation and recombination process.  
   
   
       2 . A microscope as claimed in  claim 1  which utilizes optical components which are capable of producing images in the UV band, including the range from 10 nanometers to 220 nanometers known as the vacuum ultraviolet.  
   
   
       3 . A microscope as claimed in  claim 1  which utilizes optical components which are capable of producing images in the UV including the range from 220 nanometers to 315 nanometers known as the ultraviolet C and ultraviolet B regions of the ultraviolet spectrum.  
   
   
       4 . A microscope as claimed in  claim 3  which employs an image intensifier to reduce the exposure of the sample being imaged in order to reduce effects on or damage to the sample.  
   
   
       5 . A microscope for translating spectral information to a visible color image in which light from one or more sources is separated into components by either a set of two or more filters or a device for providing wavelength limited light and then passed through or reflected off an sample then imaged onto the input of an image intensifier by an objective lens then converted to visible light by the image intensifier or other wavelength translating device the output of which is then imaged on the input of a video camera which provides the image as an electronic signal which is then converted into electronic data by a video to computer interface system and then recombined into a multicolor image by computer processing finally creating a color visible image on a display monitor where the computer is supplied with information on the position of the filters or wavelength limited light in order to synchronize acquisition of the images and the color translation and recombination process.  
   
   
       6 . A microscope as claimed in  claim 5  which employs an image intensifier to reduce the exposure of the sample being imaged in order to reduce effects on or damage to the sample.  
   
   
       7 . A microscope for translating spectral information to a visible color image in which light from a source which emits narrow spectral lines, as opposed to a continuum of spectra, is separated into components after passing through an sample and is then converted to visible polychromatic light by a converter such as an image intensifier and is then recombined into a multicolor image by a combining images captured by a video camera, video interface and computer where such images are synchronized with the filter system.  
   
   
       8 . A microscope as claimed in  claim 7  which includes an intermediate set of at least one filter which is fixed in place and only passes the illuminating light wavelength(s) from the sample by using short pass, long pass or narrow band filters.  
   
   
       9 . A microscope as claimed in  claim 8  which includes an intermediate synchronized set of at least one filter arranged to only pass the illuminating light wavelength(s) from the sample by using short pass, long pass or narrow band filters.  
   
   
       10 . A microscope as claimed in  claim 9  which includes an intermediate set of at least one filter which is fixed in place and only rejects the illuminating light wavelength(s) from the sample by using short pass, long pass or narrow band filters. The purpose of this filter set is to aid in fluorescent or autofluorescent imaging.  
   
   
       11 . A microscope as claimed in  claim 10  which includes an intermediate synchronized set of at least one filter arranged to only rejects the illuminating light wavelength(s) from the sample by using very narrow band notch filters. The purpose of such filters is to allow any Raman re-emission spectra to be viewed in the microscope.  
   
   
       12 . A microscope as claimed in  claim 11  which includes polarizing means capable of transmitting the wavelengths being used for illumination where such polarizing means are located in either or both of: between the source and the sample, or between the sample and the image intensifier and where one or both of such polarizing means are adjustable in a rotary fashion under manual or computer control.  
   
   
       13 . A microscope as claimed in  claim 12  which includes polarizing means capable of transmitting the wavelengths being used for illumination where such polarizing means are located in either or both of: between the source and the sample, or between the sample and the image intensifier, and where such polarizing means are located in synchronized filter wheels mounted on the same shaft as the sample, image or intermediate filter wheels.  
   
   
       14 . A microscope as claimed in  claim 13  which includes filters which have engineered spectral responses which compensate for differences in the intensity of illuminating spectra from the source, or differences in the spectral transmission efficiency of the optical components, or differences in the spectral photon to electron conversion efficiency of the photocathode of the image intensifier.  
   
   
       15 . A microscope as claimed in  claim 14  which includes a single or dual coaxial tubular support means, as opposed to the traditional C frame, microscope supporting structure for the purpose of vibration reduction, and the resulting image improvement, at high resolutions.  
   
   
       16 . An optical microscope system where an image intensifier and CCD camera combined with a computerized image capture and image processing system is used to convert images collected in wavelengths outside the normal range of human vision, such as soft x-ray, UV or IR, to visible images and where, while at least one of the images collected is in the range 200 nanometers to 300 nanometers, some of the other images used to produce the final color image can be collected in the range from 300 to 3300 nanometers.  
   
   
       17 . A microscope as claimed in  claim 16  where the image intensifier is a microchannel plate type image intensifier.  
   
   
       18 . A microscope as claimed in  claim 16  where the image intensifier is a diode type image intensifier.  
   
   
       19 . A microscope as claimed in  claim 18  where the camera is a tube type camera such as a vidicon, saticon or image orthicon.  
   
   
       20 . A microscope as claimed in  claim 18  where the camera is a photodiode array or photodiode array/CCD hybrid with direct UV sensitivity not requiring a secondary coating of a phosphor such as lumigen.