Microscope with a viewing direction perpendicular to the illumination direction

• US 7,554,725 B2
• Filed: 06/06/2003
• Issued: 06/30/2009
• Est. Priority Date: 12/09/2002
• Status: Expired due to Term

• Current Assignee: Europaeisches Laboratorium Fuer Molekularbiologier
• Original Assignee: Europaeisches Laboratorium Fuer Molekularbiologier
• Inventors: Huisken, Jan, Stelzer, Ernst H. K., Swoger, James H., Lindek, Steffen, Enders, Sebastian
• Primary Examiner(s): FINEMAN, LEE A
• Application Number:
  US10/538,081
• Publication Number:
  US 20060033987A1
• Time in Patent Office: 2,216 Days
• Field of Search: 359/385, 359/398, 359/368
• US Class Current: 359/385
• CPC Class Codes: G01N 21/4795 spatially resolved investigating of object in scattering medium in vivo A61B
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G02B 21/0032 Optical details of illumination, e.g. light-sources, pinholes, beam splitters, slits, fibers G02B21/0036 - G02B21/008; means for illumination of specimens in general G02B21/06
• CPC Class Codes: G02B 21/0048 scanning mirrors, e.g. rotating or galvanomirrors, MEMS mirrors
• CPC Class Codes: G02B 21/0056 based on optical coherence, e.g. phase-contrast arrangements, interference arrangements
• CPC Class Codes: G02B 21/0076 arrangements using fluorescence or luminescence
• CPC Class Codes: G02B 21/06 Means for illuminating specimens
• CPC Class Codes: G02B 21/16 adapted for ultraviolet illumination ; Fluorescence microscopes G02B21/0076 takes precedence
• CPC Class Codes: G02B 21/26 Stages; Adjusting means therefor
• CPC Class Codes: G02B 21/367 providing an output produced by processing a plurality of individual source images, e.g. image tiling, montage, composite images, depth sectioning, image comparison

Abstract

The invention relates to a microscope, in which a layer of the sample is illuminated by a thin strip of light (11) and the sample is viewed (5) perpendicular to the plane of the strip of light. The depth of the strip of light (11) thus essentially determines the depth of focus of the system. To record the image, the object (4) is displaced through the strip of light (11), which remains fixed in relation to the detector (8), and fluorescent and/or diffused light is captured by a planar detector. Objects (4) that absorb or diffuse a large amount of light are viewed from several spatial directions. The three-dimensional images, which are captured from each direction can be combined retrospectively to form one image, in which the data is weighted according to its resolution. The resolution of the combined image is then dominated by the lateral resolution of the individual images.

Claims

1. A microscope having at least one illumination beam path and at least one detection beam path, characterized in that

each illumination beam path is provided with a focusing arrangement for producing a linear object illumination region which extends in a direction of an illumination axis of the illumination beam path,

a detection direction of the at least one detection beam path is approximately orthogonal to the linear object illumination region, and

at least one mobile arrangement is provided for producing a relative movement between the linear object illumination region and an object to be studied,

wherein the linear object illumination region is essentially limited to the direction of the illumination axis.

2. The microscope as claimed in claim 1, characterized in that

the at least one mobile arrangement is designed to produce a relative movement between the object and the linear object illumination region essentially orthogonally to the illumination axis and the detection direction.

3. The microscope as claimed in claim 2, characterized in that

the at least one mobile arrangement is designed to move the object in order to produce a relative movement.

4. The microscope as claimed in claim 2, characterized in that

the at least one mobile arrangement is designed to move the at least one illumination beam path at least in the linear object illumination region provided by it, in order to produce the relative movement.

5. The microscope as claimed in claim 4, characterized in that

the at least one mobile arrangement is designed to move the at least one detection beam path in accordance with the movement of the at least one illumination beam path, at least in its region near the object.

6. The microscope as claimed in claim 1, characterized in that

the at least one detection beam path has a detector with a multiplicity of detector pixels.

7. The microscope as claimed in claim 6, characterized in that

the number and positioning of the detector pixels of the detector are selected so that the at least one detection beam path projects a section of the object, illuminated by the at least one illumination beam path in the object illumination region, essentially fully onto the detector.

8. The microscope as claimed in claim 1, characterized in that

the at least one mobile arrangement is designed to move the object to be studied essentially in the direction of the detection direction of the at least one detection beam path.