Laser microdissection apparatus and method

• US 8,715,955 B2
• Filed: 09/08/2005
• Issued: 05/06/2014
• Est. Priority Date: 09/09/2004
• Status: Active Grant

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Donovan, Brian W., Baer, Thomas M.
• Primary Examiner(s): Ford, Allison
• Assistant Examiner(s): FERNANDEZ, SUSAN EMILY
• Application Number:
  US11/222,281
• Publication Number:
  US 20060087643A1
• Time in Patent Office: 3,162 Days
• Field of Search: 356/36
• US Class Current: 435/40.5
• CPC Class Codes: G01N 1/28 Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Qmounting specimens on microscopic slides G02B21/34; means for supporting the objects or the materials to be analysed in electron microscopes H01J37/20 ; laboratory gas handling apparatus B01L5/00
• CPC Class Codes: G01N 1/2813 Producing thin layers of samples on a substrate, e.g. smearing, spinning-on G01N1/30 takes precedence
• CPC Class Codes: G01N 1/286 involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising microtomes G01N1/06; pulverising in general B02C; mixing in general B01F
• CPC Class Codes: G01N 1/30 Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
• CPC Class Codes: G01N 1/44 Sample treatment involving radiation, e.g. heat
• CPC Class Codes: G01N 2001/2826 Collecting by adsorption or absorption
• CPC Class Codes: G01N 2001/2833 Collecting samples on a sticky, tacky, adhesive surface
• CPC Class Codes: G01N 2001/284 using local activation of adhesive, i.e. Laser Capture Microdissection
• CPC Class Codes: G01N 2001/2886 Laser cutting, e.g. tissue catapult
• CPC Class Codes: Y10T 156/10 Methods of surface bonding and/or assembly therefor
• CPC Class Codes: Y10T 156/1054 and simultaneously bonding [e.g., cut-seaming]

Abstract

Systems and methods for automated laser microdissection are disclosed. In one variation, targeted biological material is manually or automatically selected and a transfer film is placed in juxtaposition to the location of an interior of a cut path. In another variation, a sample of biological material is mounted onto a polymer membrane which is then placed onto a substrate. Targeted biological material is manually or automatically selected and a transfer film is placed in juxtaposition with the targeted biological material on the side of the biological material. In yet another variation, a sample of biological material is mounted onto a polymer membrane which is then inverted onto a substrate. Targeted biological material is manually or automatically selected and a transfer film is placed in juxtaposition with the targeted biological material on the side of the polymer membrane. Then, an UV laser cuts along a cut path around the targeted portions of biological material in a closed cut path or a substantially closed cut path defining an interior and an exterior portion of the tissue sample. In a substantially closed cut path, bridges are left spanning the interior of the cut path and the exterior of the cut path. An IR laser activates at least a portion of the transfer film such that the transfer film in the vicinity of targeted portion adheres to the biological material interior to the cut path. The transfer film is then removed separating the targeted portions of biological material which are adhered to the transfer film from the remaining portion of the tissue sample.

Claims

1. A method for laser microdissection comprising:

providing a first substrate having a surface;

applying a layer of biological material to the surface of the first substrate;

providing a second substrate having a surface with a transfer film disposed thereon, wherein the transfer film has adhesive characteristics upon activation by electromagnetic energy;

identifying at least one targeted portion of biological material to be microdissected, wherein the at least one targeted portion includes desired and undesired biological material;

bringing the transfer film into juxtaposition with the first substrate on the side of the biological material in the location of the at least one targeted portion of biological material;

activating a first laser source so as to describe at least one closed or substantially closed path around the at least one targeted portion of biological material to be microdissected, wherein the first laser source is configured to erode the biological material along the described path and defining an interior and exterior;

activating a second laser source and directing the second laser source at the interior of the at least one described path so as to activate at least one region of the transfer film so that the at least one activated region of transfer film adheres to the at least one interior portion of biological material, wherein the at least one activated region of transfer film adheres to at least one portion of the undesired biological material; and

separating the second substrate with its attached transfer film and the at least one adhered targeted portion of biological material from the remaining layer of biological material.

2. The method of claim 1 wherein the first laser source is a UV laser source.

3. The method of claim 1 wherein the second laser source is an IR laser source.

4. The method of claim 1 wherein activating the first laser source includes describing a substantially closed path such that there remains at least one bridge between the interior to the exterior wherein the interior is joined to the surrounding exterior biological material at the at least one bridge.

5. The method of claim 4 wherein separating the second substrate with its attached transfer film and the at least one adhered targeted portion of biological material from the remaining layer of biological material includes breaking the at least one bridge.

6. The method of claim 4 wherein activating the second laser source includes activating the second laser source in between the location of the bridges.

7. The method of claim 1 wherein activating the second laser source includes activating the second laser source so as to describe at least one closed or partially closed curve at the interior of the at least one described path.

8. The method of claim 1 wherein activating the second laser source includes activating the second laser source more than one time in the interior of the at least one described path.

9. The method of claim 1 wherein bringing the second substrate with the transfer film into juxtaposition with the first substrate on the side of the biological material in the location of the at least one targeted portion of biological material includes contacting the transfer film to the biological material.

10. The method of claim 1 wherein bringing the second substrate with the transfer film into juxtaposition with the first substrate on the side of the biological material in the location of the at least one targeted portion of biological material includes spacing a substantial portion of the transfer film away from the biological material by a distance sufficient for promoting adhesion of the transfer film to the biological material upon activation of the transfer film by the second laser source.