Sample dishes for use in microscopy and methods of their use
Disclosed herein are sample dishes for use with microscopes that are simple to mount on a microscope and facilitate easy manipulation of tissue samples disposed thereon during imaging as well as methods of their use. A sample dish comprises an optical interface and, optionally, a support member that holds the optical interface. The optical interface of a sample dish is suitably transparent and planar such that a focal plane of a microscope can reside uniformly at or within a surface of a sample during imaging. In certain embodiments, a support member comprises a dish for holding excess fluid. In certain embodiments, a sample dish comprises separation ribs. In certain embodiments, a sample dish comprises one or more manipulation members (e.g., tabs). In certain embodiments, a sample dish is used with an imaging artifact reducing fluid.
1. A method of imaging samples using an imaging system, comprising:
providing a first sample dish comprising a substantially planar optical interface attached to a support member that is disposed at least partially around a periphery of the optical interface;
disposing the first sample dish on the imaging system such that a mounting side of the optical interface conforms to a transparent imaging window of the imaging system;
placing a sample on a sample side of the optical interface of the first sample dish, wherein the sample side of the optical interface of the first sample dish is opposite to the mounting side of the optical interface of the first sample dish;
imaging the sample using light provided through the transparent imaging window and the optical interface while the sample remains accessible to a user;
providing a second sample dish comprising a substantially planar optical interface attached to a support member that is disposed at least partially around a periphery of the optical interface;
removing the first sample dish from the imaging system;
disposing the second sample dish on the imaging system such that a mounting side of the optical interface of the second sample dish conforms to a transparent imaging window of the imaging system;
placing a second sample on a sample side of the optical interface of the second sample dish, wherein the sample side of the optical interface of the second sample dish is opposite to the mounting side of the optical interface of the second sample dish; and
imaging the second sample using light provided through the transparent imaging window and the optical interface while the sample remains accessible to a user and without adjusting a focal plane of the imaging system.
2. The method of claim 1 , wherein the support member comprises a ditch protruding from the mounting side of the optical interface that at least partially surrounds the optical interface.
3. The method of claim 2 , wherein disposing the first sample dish comprises initially disposing the first sample dish in an off-center position, wherein the ditch causes the sample dish to naturally move relative to the imaging system such that the optical interface of the sample dish comes into contact with the transparent imaging window.
4. The method of claim 1 , wherein the optical interface contacts the transparent imaging window via an imaging artifact reducing fluid disposed between, and in mutual direct contact with, the optical interface and the transparent imaging window.
5. The method of claim 1 , wherein the optical interface of the first sample dish, the optical interface of the second sample dish, or both the optical interface of the first sample dish and the optical interface of the second sample dish has a thickness between 5 μm and 500 μm.
6. The method of claim 5 , wherein the thickness is between 10 μm and 250 μm.
7. The method of claim 5 , wherein the thickness is between 25 μm and 75 μm.
8. The method of claim 5 , wherein the optical interface of the first sample dish and the optical interface of the second sample dish each have a thickness within 10 μm of a standardized thickness such that there is no need to adjust position of a focal plane of the imaging system after imaging the first sample in order to image the second sample.
9. The method of claim 8 , wherein the optical interface of the first sample dish and the optical interface of the second sample dish each have a thickness within 5 μm of the standardized thickness.
10. The method of claim 9 , wherein the optical interface of the first sample dish and the optical interface of the second sample dish each have a thickness within 2 μm of the standardized thickness.
11. The method of claim 9 , wherein the optical interface of the first sample dish and the optical interface of the second sample dish each have a diameter of at least 50 mm.
12. The method of claim 1 , wherein the optical interface of the first sample dish and the optical interface of the second sample dish each have a thickness sufficiently similar to each other such that there is no need to adjust position of a focal plane of the imaging system after imaging the first sample in order to image the second sample.
13. The method of claim 1 , wherein the optical interface of the first sample dish and the optical interface of the second sample dish have a thickness within 5 μm of each other.
14. The method of claim 1 , wherein the support member of the first sample dish and the support member of the second sample dish each comprise a ditch that surrounds a periphery of the optical interface.
15. The method of claim 14 , wherein the ditch protrudes from the mounting side of the optical interface.
16. The method of claim 15 , wherein the ditch surrounds an imaging window support of the imaging system that the transparent imaging window is attached to, when mounted.
17. The method of claim 1 , wherein the optical interface of the first sample dish and the optical interface of the second sample dish each have a dimension larger than a dimension of the transparent imaging window.
18. The method of claim 1 , wherein the first sample is laterally accessible to a user during the imaging of the first sample and the second sample is laterally accessible to the user during the imaging of the second sample.
19. The method of claim 1 , wherein the optical interface of the first sample dish and the optical interface of the second sample dish each consist essentially of imaging grade plastic.
20. The method of claim 1 , wherein (i) the first sample dish and the second sample dish are each a one-part component or (ii), for each of the first sample dish and the second sample dish, the support member is a one-part component that is attached to the optical interface.