Automated fluorescence imaging and single cell segmentation
AssigneeCanopy Biosciences, LLC
InventorChristian Henning
Life of this patent
Estimating the term…
Abstract
Systems and methods for automated, non-supervised, parameter-free segmentation of single cells and other objects in images generated by fluorescence microscopy. The systems and methods relate to both improving is image quality and to improved automatic segmentation on images. The methods will typically be performed on a digital image by a computer or processor running appropriate software stored in a memory.
Claims19
A method for calibrating an imaging system for cellular imaging, the method comprising:
- providing an imaging system for imaging cells which have tagged with a fluorophore having a range of fluorescing wavelengths;
- performing a calibration for autofluorescence comprising:
- providing said imaging system with a sample of unstained cells having a range of autofluorescing wavelengths;
- illuminating said sample with a source of illumination;
- imaging said sample across a dynamic range including all of said range of fluorescing wavelengths and all of said range of autofluorescing wavelengths;
- performing a calibration for chromatic aberration comprising:
- providing said imaging system with a sample of cells;
- illuminating said sample with a source of illumination;
- obtaining a first image of said sample of cells;
- altering a position of said imaging system relative to said sample of cells;
- and
- obtaining a new image of said sample of cells.
The method of claim herein said sample of cells is said sample of unstained cells.
A method for analyzing a cellular image, the method comprising:
- providing a sample of cells which have been tagged with a fluorophore having a range of fluorescing wavelengths;
- illuminating said sample with a source of illumination;
- imaging said illuminated sample saver said range of fluorescing wavelengths to produce a sample image;
- subtracting a calibration image from said sample image to produce a calibrated image;
- representing said image as a topological curve, a height of said curve at each curve pixel representing ate intensity of fluorescence at an image pixel in said calibrated image;
- searching said topological curve for a pixel grouping having height above a selected height;
- choosing a new height, lower than said selected height;
- repeating said searching using said new height as said selected height;
- and
- for each identified pixel grouping:
- following a convex border of said topological curve away from said pixel group to an inflection point where inflexion is decreasing at a convex side;
- and
- identifying said convex border including said pixel group as a cell in said calibrated image.
Dependencies are inferred from the claim text.