Automated microdissection instrument
View Patent ↗Systems and methods for automated laser microdissection are disclosed including automatic slide detection, position detection of cutting and capture lasers, focus optimization for cutting and capture lasers, energy and duration optimization for cutting and capture lasers, inspection and second phase capture and/or ablation in a quality control station and tracking information for linking substrate carrier or output microdissected regions with input sample or slide.
1. A method for laser microdissection, comprising:
applying a layer of biological material to a surface of a first substrate;
identifying a targeted portion of biological material located on the first substrate, wherein the targeted portion of biological material includes a desired sample for further analysis and an undesired portion of biological material;
bringing a polymer layer into juxtaposition with the first substrate on the side of the biological material in the location of the targeted portion of biological material;
activating a laser source to describe a closed or substantially closed path around the targeted portion of biological material or to deliver energy directly to the targeted portion of biological material;
transferring a portion of biological material of the targeted portion to the polymer layer by activating a capture laser directed at a region of the undesired portion included in the targeted portion, wherein the portion of biological material includes the desired sample and the undesired portion of biological material;
moving the polymer layer to a quality control station configured for viewing the portion of biological material that is present on the polymer layer, generating an image of the portion of biological material, further ablating or capturing portions of the portion of the biological material, or any combination thereof;
identifying the desired sample present on the polymer layer while the polymer layer is located in the quality control station; and
activating the laser source and directing it at the portion of biological material that is present on the polymer layer while the polymer layer is located in the quality control station.
2. The method of claim 1 wherein the step of activating the laser source while the polymer layer is located in the quality control station includes the step of ablating the undesired portion of biological material.
3. The method of claim 1 wherein said capture laser is activated by activating a second laser source.
4. The method of claim 3 wherein the second laser source is an IR laser source.
5. The method of claim 3 wherein the second laser source is a UV laser source.
6. The method of claim 1 , wherein
the polymer layer is attached to a surface of a second substrate.
7. The method of claim 6 further including separating the second substrate with its attached polymer layer and the targeted portion of biological material from the remaining layer of biological material.
8. The method of claim 6 wherein bringing the polymer layer into juxtaposition with the first substrate on the side of the biological material in the location of the targeted portion of biological material includes contacting the polymer layer, attached to the second substrate, to the biological material.
9. The method of claim 6 wherein bringing the polymer layer into juxtaposition with the first substrate on the side of the biological material in the location of the targeted portion of biological material includes spacing a portion of the polymer layer, attached to the second substrate, away from the biological material by a distance sufficient for promoting adhesion of the polymer layer to the biological material upon activation of the polymer layer by the laser source.
10. The method of claim 1 wherein the polymer layer is a transfer film having adhesive characteristics upon activation by electromagnetic energy.
11. The method of claim 1 wherein the first substrate is a polyethylene naphalate (PEN) membrane or a polyethylene terephthalate (PET) membrane.
12. The method of claim 1 :
wherein the capture laser is a second laser source and said activating a capture laser directed at a region of the undesired portion included in the targeted portion comprises activating the second laser source and directing the second laser source at an interior of the closed or substantially closed described path so as to activate at least one region of the polymer layer so that the at least one activated region of polymer layer adheres to the undesired portion of biological material.
13. The method of claim 1 wherein the laser source that describes the closed or substantially closed path around the targeted portion of biological material cuts the biological material along the described closed or substantially closed path and defines an interior and an exterior.
14. The method of claim 1 wherein the laser source that describes the closed or substantially closed path around the targeted portion of biological material is a UV laser source.
15. The method of claim 1 wherein the laser source directed at the portion of biological material that is present on the polymer layer while the polymer layer is located in the quality control station is an IR laser source.
16. The method of claim 1 wherein the step of activating the laser source that describes the closed or substantially closed path around the targeted portion of biological material 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.
17. The method of claim 16 wherein the step of transferring the portion of biological material from the layer of biological material to the polymer layer includes the step of breaking the at least one bridge.