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 relates to imaging and analyzing cell samples in a cell culture vessel by capturing a movie and generating a static range image from the movie. The static range image is composed of pixels representing the minimum fluorescence intensity subtracted from the maximum fluorescence intensity at each pixel location over a complete scan period.

Objects are defined by segmenting the static range image. The disclosed approach computes the minimum fluorescence intensity and maximum fluorescence intensity at each pixel location across a complete scan period, and the resulting per-pixel difference is used to generate the static range image, including in a microplate having a plurality of sample wells.

The disclosure further includes refining imaging and analysis over a complete scan period, including when the complete scan occurs and how the movie is formed. Additional analyses are derived from the segmented objects and from the fluorescence activity over the complete scan period, and the segmented objects are limited to one or more of a neuron, a dendrite, an axon, and a synapse.

Claims Coverage

The document includes two independent claims. Both independently cover generating a static range image using a per-pixel minimum fluorescence subtracted from a per-pixel maximum fluorescence over a complete scan period, with object definition by segmentation in one claim and a microplate context in the other.

Across both independent claims, the core inventive coverage is the per-pixel static range image computed as minimum fluorescence subtracted from maximum fluorescence across a complete scan period, derived from a captured movie, and used for segmentation-based object definition, with one claim additionally requiring a microplate with multiple sample wells.

Stated Advantages

Documented Applications