Method for laser microdissection, laser microdissection system and computer program

Patent metadata

Field	Value
Publication number	US-11874207-B2
Application number	US-202017424521-A
Country	US
Kind code	B2
Filing date	Jan 23, 2020
Priority date	Feb 5, 2019
Publication date	Jan 16, 2024
Grant date	Jan 16, 2024

Abstract

A method for laser microdissection includes: processing a microscopic examination object by a laser beam using tuples of coordinate values which respectively indicate positions of target points on the examination object at least in a first spatial direction and a second spatial direction orthogonal to the first spatial direction, positions of at least three reference points being ascertained beforehand in each case in the first and second spatial directions and also in a third spatial direction orthogonal to the first and second spatial directions; defining a reference plane based on the positions of the reference points; and determining, for the target points, further coordinate values indicating an expected position of the target points on the examination object in the third spatial direction in each case, as determined further coordinate values, the determining of the further coordinate values being performed depending on the defined reference plane.

First claim

The invention claimed is:

1. A method for laser microdissection, comprising:
processing a microscopic examination object by a laser beam using tuples of coordinate values which respectively indicate positions of target points on the examination object at least in a first spatial direction and a second spatial direction orthogonal to the first spatial direction, positions of at least three reference points being ascertained beforehand in each case in the first and second spatial directions and also in a third spatial direction orthogonal to the first and second spatial directions
defining a reference plane based on the positions of the reference points;
determining, for the target points, further coordinate values indicating an expected position of the target points on the examination object in the third spatial direction in each case, as determined further coordinate values, the determining of the further coordinate values being performed depending on the defined reference plane; and
setting a focal position of the laser beam for processing the examination object depending on the determined further coordinate values.

2. The method of claim 1 , wherein the reference plane corresponds at least partly to a plane in which a surface of the examination object is arranged.

3. The method of claim 1 , wherein the determining of the further coordinate values depending on the defined reference plane comprises in each case determining a position of the defined reference plane in the third spatial direction at a position which is indicated by an associated tuple of coordinate values in the first and second spatial directions for the respective target points.

4. The method of claim 1 , wherein an autofocus system is used, which autofocus system first is preset to a preset value and which subsequently carries out an autofocus routine based on features of the examination object, and
wherein the preset value is predefined depending on a position of the reference plane in the third spatial direction.

5. The method of claim 1 , wherein at least one of the reference points lies within or outside a region in which the examination object is situated.

6. The method of claim 5 , wherein the reference points correspond to markings on an object carrier on which the examination object is situated, or correspond to object features.

7. The method of claim 1 , wherein the tuples of coordinate values are coordinate pairs which respectively indicate positions of the target points on the examination object in the first and second spatial directions, and
wherein the determined further coordinate values are respectively assigned to the coordinate pairs.

8. The method of claim 1 , wherein the tuples of coordinate values comprise predefined coordinate values which respectively indicate a predefined position of the target points on the examination object in the third spatial direction, and
wherein the predefined coordinate values are corrected in each case based on the determined further coordinate values.

9. The method of claim 1 , wherein the coordinate values define Cartesian coordinates or polar coordinates in a referential coordinate system.

10. The method of claim 1 , wherein the target points are part of a processing line, along which the examination object is processed by the laser beam.

11. The method of claim 1 , wherein the processing of the examination object by the laser beam comprises cutting the examination object.

12. The method of claim 1 , wherein the tuples of coordinate values are ascertained in an examination device and are imported into a laser microdissection system used for processing the examination object by the laser beam.

13. A non-transitory computer-readable storage medium comprising instructions that, when executed, cause one or more processors to carry out a method according to claim 1 .

14. A laser microdissection system, comprising:
a laser having a laser beam configured to process a microscopic examination object using tuples of coordinate values which respectively indicate positions of target points on the examination object at least in a first spatial direction and a second spatial direction orthogonal to the first spatial direction,
wherein the laser microdissection system is configured to:
ascertain beforehand positions of at least three reference points at least in each case in the first and second spatial directions and in a third spatial direction orthogonal to the first and second spatial directions, to
define a reference plane based on the positions of the reference points,
determine in each case for the target points further coordinate values indicating an expected position of the target points on the examination object in the third spatial direction as determined further coordinate values,

perform the determination of the further coordinate values depending on the reference plane defined by the position of the reference point(s), and
set a focal position of the laser beam for processing the examination object depending on the determined further coordinate values.

15. The laser microdissection system of claim 14 , further comprising:
an autofocus system, which is settable to a preset value and is configured to carry out an autofocus routine based on features of the examination object,
wherein the preset value is predefinable depending on a position of the reference plane in the third spatial direction.

Assignees

• Leica Microsystems

Inventors

• Hoffmann Florian
• Schlaudraff Falk

Classifications

• G01N1/2813 Primary

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

• G01N1/286

  involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising (microtomes G01N1/06; pulverising in general B02C; mixing in general B01F) · CPC title

• G02B19/0047

  for use with a light source (G02B19/009, G02B19/0095 take precedence; details of lighting devices in general F21V; non-semiconductor lasers having optical devices external to the laser cavity H01S3/005) · CPC title

• G01N2001/284

  using local activation of adhesive, i.e. Laser Capture Microdissection · CPC title

• G01N2001/2886

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