Automated laser capture microdissection
View Patent ↗Systems and methods for automated laser capture microdissection are disclosed. High throughput microdissection is provided by using cell procurement and multi-imaging tools for pre-selecting cells of interest. Novel methods of computer-controlled cap transfer along with automated multi-slide and multi-cap placements, and automated slide and cap detection are provided. The systems and methods provide the advantages of increased speed and much lower rates of contamination.
1. A method for laser capture microdissection, comprising:
providing a laser capture instrument comprising:
a microscope having a first optical axis;
a sample-receiving stage connected to the microscope for positioning a tissue sample;
a laser selectively activable to emit a light beam along the first optical axis;
an acquisition system for capturing images along the first optical axis; and
a camera for capturing images along a second optical axis;
providing a tissue sample on the sample-receiving stage;
aligning the tissue sample and second optical axis;
capturing, with the camera, a first image of the tissue sample while positioned in the second optical axis;
identifying a region of interest of the tissue sample from the first image;
aligning the region of interest and the first optical axis;
capturing, with the acquisition system, a second image of the tissue sample while positioned in the first optical axis;
selecting at least one portion of the region of interest for microdissection;
providing a transfer film;
juxtaposing the transfer film with the at least one portion of the region of interest in the first optical axis;
activating at least one area of the transfer film adjacent to the at least one portion of the region of interest with the laser;
adhering the transfer film to the at least one portion of the region of interest that is adjacent to the at least one activated area of the transfer film; and
transferring the at least one portion of the region of interest to the transfer film.
2. The method of claim 1 wherein the step of capturing a first image of the tissue sample includes capturing a first image of the entire tissue sample.
3. The method of claim 1 wherein the step of capturing a first image of the tissue sample includes capturing within the first image, identity markings associated with the tissue sample.
4. The method of claim 1 wherein the second image is a live image.
5. The method of claim 1 wherein the first image is a static image.
6. The method of claim 1 further including the step of determining the x,y coordinates of the at least one portion of the region of interest from an analysis of the second image.
7. The method of claim 6 wherein the step of juxtaposing the transfer film with the at least one portion of the region of interest in the first optical axis includes the step of juxtaposing the transfer film with the at least one portion of the region of interest in the location of the x,y coordinates determined from the analysis of the second image.
8. The method of claim 6 further including the steps of:
providing a fluorescent excitation source; an excitation beam emitted by the fluorescent excitation source along the first optical axis having an intensity and wavelength to excite a fluorescent label associated with the tissue sample that is fluorescently labeled; and
exciting the tissue sample with the excitation beam.
9. The method of claim 8 wherein the step of determining the x,y coordinates of the at least one portion of the region of interest from an analysis of the second image includes determining the x,y coordinates of the at least one portion of the region of interest from an analysis of the second image of the tissue sample excited with the fluorescent excitation beam.
10. The method of claim 1 wherein the step of juxtaposing the transfer film includes contacting the transfer film to the tissue sample.