Confocal microscopy apparatus and method

• US 6,687,052 B1
• Filed: 09/13/2001
• Issued: 02/03/2004
• Est. Priority Date: 01/22/1999
• Status: Expired due to Term

• Current Assignee: Oxford University Innovation Limited (University of Oxford)
• Original Assignee: Isis Innovation Limited
• Inventors: Neil, Mark Andrew Aquilla, Wilson, Tony, Juskaitis, Rimvydas
• Primary Examiner(s): Robinson, Mark A.
• Assistant Examiner(s): FINEMAN, LEE A
• Application Number:
  US09/889,464
• Time in Patent Office: 873 Days
• Field of Search: 359/368, 359/388, 359/389, 359/232, 359/234, 359/235, 359/385
• US Class Current: 359/385
• CPC Class Codes: G02B 21/0024 Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
• CPC Class Codes: G02B 21/0052 Optical details of the image generation
• CPC Class Codes: G02B 21/0056 based on optical coherence, e.g. phase-contrast arrangements, interference arrangements
• CPC Class Codes: G02B 21/0068 arrangements using polarisation
• CPC Class Codes: G02B 21/0076 arrangements using fluorescence or luminescence
• CPC Class Codes: G02B 21/0084 time-scale detection, e.g. strobed, ultra-fast, heterodyne detection

Abstract

The confocal microscope has two matched light sources: a first light source (1) and a second light source (8). The light sources (1,8) are arranged to illuminate opposite sides of a modulating mask (6). The light either reflecting from or passing through the modulating mask (6) is then used to illuminate an object O supported on a mount (5). The microscope is arranged so that the object O is mounted on the opposite side of the mask (6) to a camera (7) such that light reflected from the object O passes through the modulating mask (6) before being captured by the camera (7). Subtraction of the image produced using the second light source (8) from the image produced using the first light source (1) generates a confocal image that contains substantially less noise than is possible with available confocal microscopy apparatus.

Claims

1. Confocal microscopy apparatus comprising means for directing light to a specimen;

a mask for encoding light incident on the specimen and for decoding light from the specimen, the mask having an opaque patterning on a first surface; and

one or more image detectors for detecting images of the specimen characterised in that the mask has a reflective patterning on a second surface, the reflective patterning being substantially identical to the opaque patterning on the first surface of the mask whereby one of the first and second surfaces of the mask generates a first image of the specimen consisting of a confocal image superimposed on a non-confocal image and the other of the first and second surfaces of the mask generates a second image of the specimen consisting of a non-confocal image from which a confocal image has been removed.

2. Confocal microscopy apparatus as claimed in claim 1, wherein there is further provided analysing means for generating a confocal image of the specimen using the first and second images.

3. Confocal microscopy apparatus as claimed in claim 1, further including one or more light sources.

4. Confocal microscopy apparatus as claimed in claim 3, wherein two light sources are provided to illuminate each side of the mask respectively.

5. Confocal microscopy apparatus as claimed in claim 4, wherein a second image detector detected the first and second images.

6. Confocal microscopy apparatus as claimed in claim 4, further including a second image detector for detecting the second image, the first and second image detectors being located on opposite sides of the mask.

7. Confocal microscopy apparatus as claimed in claim 1, wherein the one or more image detectors are CCD cameras.

8. Confocal microscopy apparatus as claimed in claim 1, wherein the first surface of the mask is patterned with equidistant light transmissive stripes and light barring stripes.

9. Confocal microscopy apparatus as claimed in claim 1, wherein the first and second surfaces of the mask have a random patterning.

10. Confocal microscopy apparatus as claimed in claim 1, wherein the mask is mounted for rotation about the optical axis of the apparatus.

11. A confocal microscopy method comprising providing illumination from a first light source;

encoding light incident on a specimen and decoding light from the specimen by means of a mask having an opaque patterning on a first surface; and

detecting Images of the specimen characterised by the mask has a reflective patterning on a second surface that is substantially identical to the opaque patterning on the first surface of the mask and generating a first image of the specimen consisting of a confocal image superimposed on a non-confocal image by means of one of the first or second surfaces of the mask and generating a second image of the specimen consisting of a non-confocal image from which a confocal Image has been removed by means of the other of the first or second surfaces of the mask.

12. A confocal microscopy method as claimed in claim 11, further comprising extracting a confocal image from said first and second images.

13. A confocal microscopy method as claimed in claim 12, wherein the second Image is subtracted from the first Image to extract the confocal image.

14. A confocal microscopy method as claimed in claim 11, wherein the mask is rotated.