28. MEASURING CELLULAR CALCIUM SIGNALS USING OPTICAL FLOW TECHNIQUES

Department: Bioengineering
Faculty Advisor(s): Gabriel Silva

Primary Student
Name: Marius Octavian Buibas
Email: mbuibas@ucsd.edu
Phone: 858-822-2835
Grad Year: 2010

Abstract
Calcium signaling is a valuable indicator of function in the nervous system because readily measurable transient changes in intracellular calcium concentrations in both neurons and glia are intermediary steps in most signaling pathways. An optical flow gradient algorithm was applied to fields of neurons and glia imaged using fluorescence optical microscopy in order to map functional calcium signaling with single pixel resolution. Optical flow estimates the direction and speed of motion of objects in an image between subsequent frames in a recorded digital sequence of images. Vector fields were produced for recorded image sequences (i.e. movies) that tracked the spatio-temporal magnitude of measured calcium signaling patterns. We first describe the conversion between algorithm-derived vector fields and the underlying cellular changes in calcium signaling that produce them, which allows the interpretation of the method’s outputs and results within an appropriate and meaningful neurobiological context. We then demonstrate the use of the algorithm to three example movies of in vitro network signaling using three different cell types, rMC-1 cells (a cell line established from neural retinal Muller glial cells), primary spinal cord astrocytes, and primary hippocampal neurons. Finally, we illustrate two example applications of optical flow vectors, region classification of spatiotemporal intercellular signaling patterns and pathline tracing of intracellular flows.

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