Analysis of embedded tissue samples using fluorescence-based detection
AssigneeAgilent Technologies, Inc.
InventorsVictor Lim, Adrienne McCampbell, Qingyu Wang
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Abstract
The present disclosure is directed to an improved methods and systems using autofluorescence of naturally-occurring components in a sample embedded in an embedding medium. Methods and systems are provided for determining an amount of tissue or cell preparation exposed at a surface of an embedded sample, and for preparing a tissue specimen comprising a region of interest (ROI) from an embedded sample. Methods and systems are also provided for imaging a sample of a biological tissue, and for identifying different cell types in an embedded tissue sample.
Claims11
A method of determining an amount of tissue or cell preparation exposed at a surface of a sample embedded in an embedding medium comprising:
- irradiating the embedded tissue or cell preparation sample at a wavelength which causes endogenous components of the tissue to autofluoresce;
- obtaining an image of the autofluorescence emitted by the embedded tissue or cell preparation sample;
- and
- determining a percentage of the image at the surface of the embedding medium which is occupied by tissue or cell preparation;
- wherein the percentage of the image at the surface of the embedding medium which is occupied by tissue is determined by:
- slicing a tissue section from the embedded tissue sample, wherein the embedded tissue sample comprises the tissue and the embedding medium;
- irradiating the embedded tissue sample with electromagnetic radiation having an excitation wavelength;
- generating a fluorescence image from an autofluorescence emission of the embedded sample;
- determining a local focus measure for pixels of the fluorescence image;
- constructing a depth map of the tissue based on evaluating image blur of the fluorescence;
- and
- determining a sectioning plane for the embedded sample, based on the depth map.
Dependencies are inferred from the claim text.