Automated microdissection instrument with controlled focusing during movement of a laser beam across a tissue 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 optimizing the focus of a laser beam for processing a biological sample comprising the steps of:
mounting a tissue sample on a first side of a worksurface of a laser capture microdissection (LCM) instrument, wherein the worksurface intersects a primary optical axis of an optical system of the LCM instrument;
focusing a first objective lens of the optical system of the LCM instrument on the tissue sample mounted on the first side worksurface to form a clear image of the tissue sample acquired by an image acquisition system, wherein the first objective lens is in the primary optical axis below a second side of the worksurface;
moving a laser beam focused through a laser focusing lens from a laser source for processing the tissue sample to an area of the tissue sample including a non-targeted biological material, wherein the laser source and the laser focusing lens are in the primary optical axis above the first side of the worksurface;
focusing the laser beam directed at non-targeted biological material to a discrete point by adjusting the laser focusing lens to a baseline setting defining a distance of the laser focusing lens to a focal plane;
re-focusing the first objective lens on the tissue sample by moving the first objective lens a distance in response to an a user navigating across the tissue sample; and
keeping the laser source at a first desired focus by moving the laser focusing lens from the baseline setting by substantially the same distance that the first objective lens was moved when re-focused.
2. The method of claim 1 , wherein if the first objective lens is moved toward the worksurface on re-focus, the step of keeping the laser source at the first desired focus includes moving the laser focusing lens away from the worksurface and if the first objective lens is moved away from the worksurface on re-focusing, the step of automatically keeping the laser source at the first desired focus includes moving the laser focusing lens toward the worksurface.
3. The method of claim 1 , further including changing the first objective lens to a second objective lens.
4. The method of claim 3 , further including:
focusing the second objective lens after the changing the first objective lens; and
maintaining the focusing lens at the first desired focus of the first objective lens using an automatic focus lock.
5. The method of claim 4 , further including the steps of:
automatically re-focusing the second objective lens on the tissue sample by moving the second objective lens a distance;
automatically keeping the laser source at a second desired focus by moving the focusing lens by the same distance that the second objective lens was moved when re-focused.
6. The method of claim 1 , wherein the laser source is an IR laser.
7. The method of claim 6 , further comprising activating the IR laser to capture a sample of targeted biological material.
8. The method of claim 1 , wherein the laser source is a UV laser.
9. The method of claim 8 , further comprising activating the UV laser to cut a sample of targeted biological material.
10. The method of claim 1 , wherein the image acquisition system is a camera.