Preparation of tissue sections using fluorescence-based detection

Inventors

Schleifer, Kyle • Bernick, Kristin Briana • Mccampbell, Adrienne • Sampas, Nicholas M. • Lim, Victor

Assignees

Agilent Technologies Inc

Abstract

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.

Core Innovation

The invention relates to distinguishing tissue from an embedding medium in an embedded tissue sample by using fluorescence. The approach irradiates an embedded tissue sample with light having a wavelength of from about 200 nm to about 600 nm, where the tissue is optically indistinguishable from the embedding medium under normal lighting conditions. After irradiation, the method analyzes endogenous fluorescence emission of the embedded tissue sample to differentiate the tissue from the embedding medium.

A further aspect of the invention distinguishes tissue from the embedding medium by analyzing endogenous fluorescence emission attributed to tissue autofluorescence. The endogenous fluorescence emission is produced by tissue autofluorescence of a tissue component, including elastin or collagen, enabling optical differentiation in situations where tissue and embedding medium appear the same under normal or visible lighting. The method also contemplates an embedded tissue sample comprising paraffin or a formalin-fixed paraffin-embedded tissue block.

Another aspect provides that the irradiation and fluorescence analysis support identifying where the tissue is located in the embedded sample. The invention uses image-based analysis, including determining the location of tissue by using pixel intensity from a digital image, and can further determine the location of a tissue surface. After determining the tissue surface location, the invention includes slicing or trimming tissue sections, as part of the described workflow.

Claims Coverage

The document provides two independent methods that distinguish tissue from an embedding medium using fluorescence. Across these independent claims, the main inventive features are two: analyzing endogenous fluorescence emission of the embedded sample to differentiate tissue from embedding medium when they are optically indistinguishable under normal lighting, and distinguishing by analyzing endogenous fluorescence emission of the embedding medium specifically while irradiating with 200–600 nm light.

Both independent claims define a 200–600 nm irradiation step followed by analyzing endogenous fluorescence emission to distinguish tissue from embedding medium despite optical indistinguishability under normal or visible lighting. The first claim analyzes endogenous fluorescence emission of the embedded sample broadly, while the second claim analyzes endogenous fluorescence emission of the embedding medium.

Stated Advantages

Documented Applications

No documented applications found