Two-photon calcium imaging from head-fixed Drosophila during optomotor walking behavior.

Drosophila melanogaster is a model organism rich in genetic tools to manipulate and identify neural circuits involved in specific behaviors. Here we present a technique for two-photon calcium imaging in the central brain of head-fixed Drosophila walking on an air-supported ball. The ball's motion is tracked at high resolution and can be treated as a proxy for the fly's own movements. We used the genetically encoded calcium sensor, GCaMP3.0, to record from important elements of the motion-processing pathway, the horizontal-system lobula plate tangential cells (LPTCs) in the fly optic lobe. We presented motion stimuli to the tethered fly and found that calcium transients in horizontal-system neurons correlated with robust optomotor behavior during walking. Our technique allows both behavior and physiology in identified neurons to be monitored in a genetic model organism with an extensive repertoire of walking behaviors.

g-PRIME: A Free, Windows Based Data Acquisition and Event Analysis Software Package for Physiology in Classrooms and Research Labs.

We present g-PRIME, a software based tool for physiology data acquisition, analysis, and stimulus generation in education and research. This software was developed in an undergraduate neurophysiology course and strongly influenced by instructor and student feedback. g-PRIME is a free, stand-alone, windows application coded and "compiled" in Matlab (does not require a Matlab license). g-PRIME supports many data acquisition interfaces from the PC sound card to expensive high throughput calibrated equipment. The program is designed as a software oscilloscope with standard trigger modes, multi-channel visualization controls, and data logging features. Extensive analysis options allow real time and offline filtering of signals, multi-parameter threshold-and-window based event detection, and two-dimensional display of a variety of parameters including event time, energy density, maximum FFT frequency component, max/min amplitudes, and inter-event rate and intervals. The software also correlates detected events with another simultaneously acquired source (event triggered average) in real time or offline. g-PRIME supports parameter histogram production and a variety of elegant publication quality graphics outputs. A major goal of this software is to merge powerful engineering acquisition and analysis tools with a biological approach to studies of nervous system function.

A polyimide pressure-contact multielectrode array for implantation along a submillimeter neural process in small animals.

We describe a microelectrode system for recordings from nerve bundles with diameters ranging from 20-200 microm. A novel polyimide structure allows for a planar microfabricated device to constrain a free neural process against several recording sites. This polyimide array contains multiple zigzag vias through which a small nerve process may be woven while remaining functionally intact in a live preparation. Our electrode construct features the benefits of nerve cuffs (evenly spaced electrodes in a polymer) and the functionality of a nerve hook (ability to connect to submillimeter processes). The device records extracellular action potentials in the red-swamp crayfish, Procambarus clarkii. Action potential propagation is monitored at several sites along a constrained nerve in this model organism's peripheral nervous system. Details of temporal accuracy and error sources in absolute conduction velocity measurements from microelectrode arrays are discussed.

An inexpensive sub-millisecond system for walking measurements of small animals based on optical computer mouse technology.

Stimuli from a broad spectrum of sensory modalities, including visual, auditory, thermal, and chemical, can elicit walking responses in animals, reflecting neural activity in sensorimotor pathways. We have developed an integrated walking measurement system with sub-millisecond temporal accuracy capable of detecting position changes on the order of 100 microm. This tracking system provides the experimenter with a means by which to map out the response spectrum of a tethered animal to any number of sensory inputs on time scales relevant to propagation in the nervous system. The data acquisition system consists of a modified optical computer mouse, a microcontroller with peripheral support circuitry, a binary stimulus sync line, and a serial (RS-232) data transfer interface. The entire system is constructed of relatively inexpensive components mostly converted from commercially available peripheral devices. We have acquired walking data synchronized with auditory stimuli at rates in excess of 2100 samples per second while applying this system to the walking phonotactic response of the parasitic fly Ormia ochracea.

Direct Orthogonal Distance to Quadratic Surfaces in 3D.

Discovering the orthogonal distance to a quadratic surface is a classic geometric task in vision, modeling, and robotics. I describe a simple, efficient, and stable direct solution for the orthogonal distance (foot-point) to an arbitrary quadratic surface from a general finite 3D point. The problem is expressed as the intersection of three quadratic surfaces, two of which are derived from the requirement of orthogonality of two non-coincident planes with the tangent plane to the quadric. A sixth order single-variable polynomial is directly generated in one coordinate of the surface point. The method detects intersection points at infinity and operates smoothly across all real quadratic surface classes. The method also geometrically detects continuums of orthogonal points (i.e., from the exact center of a sphere). I discuss algorithm performance, compare it to a state-of-the-art estimator, demonstrate the algorithm on synthetic data, and describe extension to arbitrary dimension.

Real-time development of data acquisition and analysis software for hands-on physiology education in neuroscience: G-PRIME.

We report on the real-time creation of an application for hands-on neurophysiology in an advanced undergraduate teaching laboratory. Enabled by the rapid software development tools included in the Matlab technical computing environment (The Mathworks, Natick, MA), a team, consisting of a neurophysiology educator and a biophysicist trained as an electrical engineer, interfaced to a course of approximately 15 students from engineering and biology backgrounds. The result is the powerful freeware data acquisition and analysis environment, "g-PRIME." The software was developed from week to week in response to curriculum demands, and student feedback. The program evolved from a simple software oscilloscope, enabling RC circuit analysis, to a suite of tools supporting analysis of neuronal excitability and synaptic transmission analysis in invertebrate model systems. The program has subsequently expanded in application to university courses, research, and high school projects in the US and abroad as free courseware.