Preparation of tissue sections using fluorescence-based detection
The present disclosure is directed to an improved method for distinguishing tissue from an embedding medium, such as paraffin in a formalin-fixed paraffin-embedded sample. The method involves the use of fluorescence of naturally-occurring species in tissue to determine the location of the tissue in the embedded sample. An embedded sample is generally excited by light of a selected wavelength, and the fluorescence emission at an emitted wavelength is used to locate the boundary or location of the tissue in the embedded sample.
1. An apparatus for slicing a tissue section from an embedded sample, the apparatus comprising:
a microtome comprising a sample holder;
at least one light source directed at the sample holder, wherein the at least one light source is selected from the group consisting of an LED light source, a mercury arc lamp, a Xenon arc lamp, and a laser; and
an optical system positioned to capture endogenous emitted light from a tissue located in an embedded sample held by the sample holder.
2. The apparatus of claim 1 , further comprising a processor in communication with the optical system and configured to provide a signal based on an endogenous fluorescence emission from the sample and to determine a location of the tissue in the embedded sample based on the endogenous fluorescence emission.
3. The apparatus of claim 1 , wherein the at least one light source is an LED light source.
4. The apparatus of claim 1 , wherein the at least one light source is a mercury arc lamp.
5. The apparatus of claim 1 , wherein the at least one light source is a Xenon arc lamp.
6. The apparatus of claim 1 , wherein the at least one light source is a laser.
7. The apparatus of claim 1 , further comprising a dichroic beam splitter.
8. The apparatus of claim 7 , wherein the dichroic beam splitter is part of a filter cube.
9. The apparatus of claim 8 , wherein the filter cube comprises an emission filter or an excitation filter.
10. The apparatus of claim 1 , further comprising one or more excitation filters.
11. The apparatus of claim 1 , further comprising one or more emission filters.
12. The apparatus of claim 1 , further comprising two or more emission filters, and at least one mechanism for switching between said two or more emission filters.
13. The apparatus of claim 1 , wherein the apparatus comprises at least two light sources.
14. The apparatus of claim 1 , wherein the apparatus comprises at least three light sources.
15. The apparatus of claim 1 , wherein the apparatus comprises at least four light sources.
16. The apparatus of claim 1 , wherein the optical system comprises a fluorescence microscope or a camera.
17. The apparatus of claim 1 , wherein the optical system comprises a digital camera.
18. The apparatus of claim 1 , wherein the apparatus comprises at least two different light sources.