Live cell visualization and analysis

Inventors

APPLEDORN, Daniel • Endsley, Eric • Holtz, Nevine • Neagle, Brad • Rock, David • Schroeder, Kirk

Assignees

Sartorius Bioanalytical Instruments Inc

Abstract

Systems and methods are provided for automatically imaging and analyzing cell samples in an incubator. An actuated microscope operates to generate images of samples within wells of a sample container across days, weeks, or months. A plurality of images is generated for each scan of a particular well, and the images within such a scan are used to image and analysis metabolically active cells in the well. Tins analysis includes generating a “range image” by subtracting the minimum intensity value, across the scan, for each pixel from the maximum intensity value. This range image thus emphasizes cells or portions of cells that exhibit changes in activity over a scan period (e.g., neurons, myocytes, cardiomyocytes) while de-emphasizing regions that exhibit consistently high intensities when images (e.g., regions exhibiting a great deal of autofluorescence unrelated to cell activity).

Core Innovation

The invention identifies biologically active cells during one or more periods of time by generating a plurality of fluorescence activity images of a sample contained within a sample container. Each image includes pixel values corresponding to pixel locations within an image frame, and the invention generates a fluorescence range image by determining, for each pixel location, a range of a set of pixel values across the plurality of fluorescence activity images.

Based on the fluorescence range image, the invention determines, relative to the image frame, a location of one or more active objects within the sample container during a corresponding period of time. Each active object is a portion of at least one respective biologically active cell present within the sample during the corresponding period of time, and for each active object the invention determines a respective time-varying activity level across the corresponding period of time.

In one implementation, the fluorescence range image pixel locations corresponding to the location of a given active object are used so that the time-varying activity level is determined based on a set of pixel values that have pixel locations proximate to the determined location of the active object. Based on the time-varying activity levels determined for the one or more active objects, the invention selects a subset of active objects, where selecting a particular active object includes determining that the time-varying activity level exhibits at least one burst.

Claims Coverage

The provided material includes three independent claims. Each independent claim centers on identifying biologically active cells from fluorescence activity images across one or more periods of time, deriving active object locations, and determining time-varying activity levels; one adds selecting a subset of active objects based on burst behavior, and one specifies how activity levels are determined using pixel values proximate to each active object location.

Across the independent claims, the inventive coverage focuses on deriving a fluorescence range image to locate active objects in an image frame and determining time-varying activity levels for those active objects from fluorescence image pixel values, including proximate pixel sets. One independent claim further covers selecting a subset of active objects by determining that a time-varying activity level exhibits at least one burst.

Stated Advantages

Documented Applications

No documented applications found