Method for defining a laser microdissection region, and associated laser microdissection system

Patent metadata

Field	Value
Publication number	US-9804144-B2
Application number	US-201314049241-A
Country	US
Kind code	B2
Filing date	Oct 9, 2013
Priority date	Oct 9, 2012
Publication date	Oct 31, 2017
Grant date	Oct 31, 2017

Abstract

A method for laser microdissection of a laser microdissection region of a prepared specimen includes driving a holder for the specimen into a holding position using a control device. First and second digital images are captured that depict a same portion of the prepared specimen, with the first image depicting the portion under at least one first microscopic examination method and the second image depicting the portion under at least a second microscopic examination method. A live overlay image is generated of the portion of the prepared image in a live mode. The live overlay is presented on a display area with the images overlaid onto one another. A marking is generated and captured on the live overlay image so as to define the laser microdissection region.

First claim

What is claimed is:

1. A method for defining a laser microdissection region of a prepared specimen that is held by a holder of a laser microdissection system, the method comprising:
driving the holder into a holding position using a control device of the laser microdissection system;
capturing a first digital image that depicts a portion of the prepared specimen, the first digital image depicting the portion of the prepared specimen under at least one first microscopic examination method;
capturing a second digital image of the same portion of the prepared specimen, the second digital image depicting the portion of the prepared specimen under at least a second microscopic examination method;
generating a live overlay image of the same portion of the prepared specimen by overlaying the first digital image and the second digital image onto one another in a live mode;
presenting the live overlay image on a display area; and
generating and capturing a marking on the live overlay image so as to define the laser microdissection region.

2. The method according to claim 1 , wherein the holder is immobilized in the holding position so as to retain the prepared specimen before the capturing the first digital image and kept immobilized in the holding position during the capturing the first digital image and the second digital image and until after the capturing the marking on the live overlay image.

3. The method according to claim 1 , further comprising cutting out, by laser microdissection, the laser microdissection region in accordance with the marking on the live overlay image.

4. The method according to claim 3 , wherein the holder is immobilized in the holding position so as to retain the prepared specimen before the capturing the first digital image and kept in the holding position during the capturing the first digital image and the second digital image and until after the cutting out, by laser microdissection, the laser microdissection region.

5. The method according to claim 1 , wherein the first digital image and the second digital image are overlaid in pixel-precise fashion.

6. The method according to claim 1 , wherein the first digital image and the second digital image capture the same portion of the prepared specimen at the same magnification.

7. The method according to claim 1 , where the first digital image and the second digital image capture the same portion of the prepared specimen at different magnifications, and where for generation of the live overlay image, the one of the first digital image and the second digital image having, the lower magnification is electronically zoomed to the higher magnification.

8. The method according to claim 1 , further comprising
importing the first digital image into a calculation unit of the control device, the importing the first digital image including importing local coordinates of the prepared specimen relative to the laser microdissection system upon acquisition of the first digital image, and
wherein the holding position is driven to on the basis of the local coordinates.

9. The method according to one claim 1 , further comprising at least one of:
storing the first digital image,
storing the at least one second digital image,
storing the overlay image, or
storing local coordinates of the holding position.

10. The method according to claim 1 , further comprising capturing a further digital image,
wherein the further digital image depicts the portion of the prepared specimen under a further type of illumination, and
wherein the first digital image, the second digital image, and the further digital image are overlaid onto one another in the overlay image.

11. The method according to claim 1 , wherein the first digital image is a second overlay image in which a third digital image and a fourth digital image are overlaid onto one another, where the third and the fourth digital image depict the portion of the prepared specimen under different types of illumination.

12. A tangible non-transient computer-readable medium having computer-executable instructions stored thereon for defining a laser microdissection region of a prepared specimen that is held by a holder of a laser microdissection system, the computer-executable instructions including instructions for:
driving the holder into a holding position using a control device of the laser microdissection system;
capturing a first digital image that depicts a portion of the prepared specimen, the first digital image depicting the portion of the prepared specimen under at least one first microscopic examination method;
capturing a second digital image of the same portion of the prepared specimen, the second digital image depicting the portion of the prepared specimen under at least a second microscopic examination method;
generating a live overlay image of the same portion of the prepared specimen by overlaying the first digital image and the second digital image onto one another in a live mode;
presenting the live overlay image on a display area; and
generating and capturing a marking on the live image so as to define the laser microdissection region.

13. A laser microdissection system comprising:
a holder for a prepared specimen;
a control device for driving the holder into a holding position;
a microscope for capturing a portion of the prepared specimen in the holding position under a first and at least one second microscopic examination method;
a digital image capture device for capturing a first and a second digital image of the prepared specimen portion, the first digital image being generated under the first microscopic examination method, and the second digital image being generated under the at least one second microscopic examination method, the digital image capture device including a calculation unit configured so as to overlay the first and the second digital images in a live mode and present the overlaid first and second digital images on a display area as a live overlay image;
a marking capture unit configured to capture a marking, generated on the live overlay image, for defining the laser microdissection region within the prepared specimen portion; and
a laser device configured to dissect a laser microdissection region by laser-cutting the region out of the prepared specimen portion along the marking.

14. The laser microdissection system according to claim 13 , wherein the digital image capture device includes a temporary memory for storing the first and/or the second digital image, as well as the overlay image.

15. The laser microdissection system according to claim 14 , wherein the temporary memory and the calculation unit of the image capture device and/or the marking capture unit is each a constituent of the control device and/or of an associated computer.

16. The method according to claim 1 , wherein the portion of the prepared specimen is located at the same position for the capturing of the first digital image as for the capturing of the second digital image.

17. The tangible non-transient computer-readable medium of claim 12 , wherein the holder is immobilized in the holding position before the first digital image and/or the second digital image is captured, and the holder is kept immobilized until after capture of t:he marking.

18. The tangible non-transient computer-readable medium of claim 12 , wherein the portion of the prepared specimen is located at the same position for the capturing of the first digital image as for the capturing of the second digital image.

19. The laser microdissectio

Assignees

• Leica Microsystems

Inventors

• Schlaudraff Falk
• Hoffmann Florian

Classifications

• G01N2001/2886

  Laser cutting, e.g. tissue catapult · CPC title

• G01N2001/282

  with mapping; Identification of areas; Spatial correlated pattern · CPC title

• G01N1/2813 Primary

  Producing thin layers of samples on a substrate, e.g. smearing, spinning-on (G01N1/30 takes precedence) · CPC title

• G01N33/483 Primary

  Physical analysis of biological material · CPC title