Automated microdissection instrument and method for processing a biological sample
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 processing a biological sample using a laser microdissection instrument, comprising:
calibrating a capture laser to determine an optimal setting for wetting a transfer film mounted on a cap, said calibration comprising:
firing the capture laser at a selected test fire location on the cap;
recording intensity data of a test image at the selected test fire location; and
determining whether wetting of the transfer film at the selected location is optimal based on detecting a maximum intensity difference between adjacent pixels;
bringing the transfer film into juxtaposition with a targeted portion of biological material on a substrate;
directing the capture laser to fire at the targeted portion of biological material to form an area of adhesion between a wetted portion of the transfer film and the targeted portion of biological material; and
moving the cap away from the substrate, thereby removing the targeted portion of biological material with the cap.
2. The method of claim 1 , wherein calibrating the capture laser further comprises:
adjusting a capture laser firing parameter in response to determining that wetting of the transfer film at the selected test fire location is not optimal.
3. The method of claim 2 , wherein adjusting the capture laser firing parameter comprises adjusting a power setting parameter of the capture laser.
4. The method of claim 2 , wherein adjusting the capture laser firing parameter comprises adjusting a duration of firing parameter of the capture laser.
5. The method of claim 2 , wherein the method further comprises repeating the steps of firing, recording, determining and adjusting until determining wetting of a selected test fire location is optimal.
6. The method of claim 1 , wherein calibrating the capture laser further comprises:
creating a calibration matrix for a test firing pattern by entering by an end user:
a number of steps for power increments, a power increment per steps, and a distance between each of a selected test fire location to set up a test firing pattern for a first axis;
a number of steps for duration increments, a duration increment per step, and a distance between each of a selected test fire location to set up a second test firing pattern on a second axis; and
firing the capture laser according to the calibration matrix to provide an array of test fire locations; and
selecting an optimal power setting and an optimal duration for wetting a transfer film from a test fire location in the array of test fire locations.
7. The method of claim 1 , the method further comprises:
calibrating a cutting laser to determine an optimal setting for ablation of biological material on a substrate;
imaging the cap in a quality control station; and
ablating additional biological material from the targeted portion of biological material in response to determining that removal of additional biological material is indicated.
8. The method of claim 7 , wherein calibrating the capture laser further comprises:
firing the cutting laser at a selected test fire location on an area of biological material;
determining whether ablation of the biological material at the selected test fire location is optimal; and
adjusting a cutting laser firing parameter in response to determining that ablation of the biological material at the selected test fire location is not optimal.
9. The method of claim 8 , wherein the method further comprises repeating the steps of firing, determining and adjusting until determining ablation of biological material at a selected test fire location is optimal.
10. The method of claim 8 , wherein adjusting the cutting laser firing parameter comprises adjusting a power setting parameter of the cutting laser.
11. The method of claim 8 , wherein adjusting the cutting laser firing parameter comprises adjusting a duration of firing parameter of the cutting laser.
12. A method for calibrating a laser microdissection instrument, comprising:
firing a capture laser at a selected test fire location a transfer film mounted on a cap;
recording intensity data of a test image at the selected test fire location; and
determining whether wetting of the transfer film at the selected location is optimal based on detecting a maximum intensity difference between adjacent pixels.
13. The method of claim 12 further comprising:
adjusting a capture laser firing parameter in response to determining that wetting of the transfer film at the selected test fire location is not optimal.
14. The method of claim 13 , wherein adjusting the capture laser firing parameter comprises adjusting a power setting parameter of the capture laser.
15. The method of claim 13 , wherein adjusting the capture laser firing parameter comprises adjusting a duration of firing parameter of the capture laser.
16. The method of claim 13 , wherein the method further comprises repeating the steps of firing, recording, determining and adjusting until determining wetting of a selected test fire location is optimal.