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 sample dish mounted on an imaging system for imaging a sample, the sample dish comprising:
a support member; and
a substantially planar transparent optical interface having a sample side and a mounting side opposite to the sample side, wherein the optical interface is attached to the support member and the support member is disposed at least partially around a periphery of the optical interface,
wherein (i) the imaging system comprises a transparent imaging window, (ii) the mounting side of the optical interface is disposed on the transparent imaging window, (iii) during the imaging the sample is disposed on the sample side of the optical interface such that the sample remains accessible to a user while light is transmitted through the transparent imaging window and the optical interface to and from the sample, and (iv) wherein a thickness of the optical interface is within 10 μm of a standardized thickness for the imaging system such that there is no need to adjust position of a focal plane of the imaging system prior to imaging,
wherein the sample dish is one of a set of sample dishes each comprising a respective support member and a respective substantially planar transparent optical interface having a respective sample side and a respective mounting side opposite to the sample side, wherein (i) the respective optical interface is attached to the respective support member and the respective support member is disposed at least partially around a periphery of the respective optical interface and (ii) a thickness of the respective optical interface is within 10 lam of the standardized thickness.
2. The sample dish of claim 1 , wherein the imaging system has a fixed focal plane and the standardized thickness corresponds to the fixed focal plane such that the fixed focal plane intersects the sample during the imaging.
3. The sample dish of claim 2 , wherein the thickness of the optical interface is such that a surface or interior of the sample falls within the fixed focal plane during the imaging.
4. The sample dish of claim 1 , wherein the thickness of the optical interface is within 5 μm of the standardized thickness.
5. The sample dish of claim 1 , wherein the thickness of the optical interface is between 5 μm and 500 μm.
6. The sample dish of claim 5 , wherein the thickness is between 10 μm and 250 μm.
7. The sample dish of claim 6 , wherein the thickness is between 25 μm and 75 μm.
8. The sample dish of claim 1 , wherein the optical interface has a dimension larger than a dimension of the transparent imaging window.
9. The sample dish of claim 1 , wherein only the optical interface contacts the microscope while the sample dish is mounted on the microscope.
10. The sample dish of claim 1 , wherein the optical interface is flexible.
11. The sample dish of claim 10 , wherein the optical interface has conformed to the transparent imaging window such that the optical interface is in contact with the transparent imaging window.
12. The sample dish of claim 1 , wherein the optical interface has a diameter of at least 50 mm.
13. The sample dish of claim 1 , wherein the optical interface has a diameter of at least 100 mm.
14. The sample dish of claim 1 , wherein a variability in thickness of the respective optical interface between ones of the set of sample dishes is less than 5 μm.
15. The sample dish of claim 1 , wherein the sample dish is a one-part component or the support member is a one-part component that is attached to the optical interface.
16. A sample characterization system for imaging the sample, the system comprising the imaging system and the sample dish of claim 1 mounted on the imaging system.
17. The sample characterization system of claim 16 , further comprising the sample disposed on the sample side of the optical interface.
18. A sample dish mounted on an imaging system for imaging a sample, the sample dish comprising:
a support member; and
a substantially planar transparent optical interface having a sample side and a mounting side opposite to the sample side, wherein (i) the optical interface is attached to the support member and the support member is disposed at least partially around a periphery of the optical interface, (ii) the optical interface is flexible, and (iii) a surface of the sample side of the optical interface varies in height by less than 10 μm across an area corresponding to a field of view of the imaging system,
wherein (a) the imaging system comprises a transparent imaging window, (b) the mounting side of the optical interface is disposed on the transparent imaging window and wherein the optical interface has conformed to the transparent imaging window such that the optical interface is in contact with the transparent imaging window, (c) during the imaging the sample is disposed on the sample side of the optical interface such that the sample remains accessible to a user while light is transmitted through the transparent imaging window and the optical interface to and from the sample, and (d) wherein a thickness of the optical interface is within 30 μm of a standardized thickness for the imaging system such that there is no need to adjust position of a focal plane of the imaging system prior to imaging.