40155-2A-RFP - 3D Acoustic Optical Deflector Based Random Access Multi-Photon Microscope System

Statement of Requirements:
The proposed instrument is a 3D Acoustic Optical Deflector (AOD) Based Random Access Multi-Photon (RAMP) Microscope System that can be used to collect data from custom defined positions at high temporal precision. The primary use of the system will be to monitor synaptic signals and action potentials from subcellular neuronal compartments and large number of neuronal somas. In addition to monitoring physiological signals, the system will also be used for stimulating neuronal elements through optogenetic methods and uncaging. The system must be able to perform two-photon RAMP imaging and laser photo-activation system to execute neuroscience experiments compatible with live cell imaging, voltage imaging and synchronized electrophysiology in brain slices in vitro and in in vivo experimental designs. The system should be optimized for fast imaging from small custom defined Region of Interest (ROIs) while simultaneously allowing experimental manipulations (i.e. uncaging). In order to achieve the requirements stated in this Request for Proposal, the proposed 3D Acoustic Optical Deflector (AOD) Based Random Access Multi-Photon (RAMP) Microscope System will be combined with lasers that will be purchased separately in coordination with the successful proponent. The successful proponent will be responsible for integrating the lasers into the microscope system

General Description:
The objective of the research is to understand the physiological processes by which the brain translates incoming information and code new memories. In order to achieve this we need to image neuronal activity at a very high speed. The microscope system will be used to monitor neuronal voltage signals at high temporal and spatial resolution. Data obtained with the new imaging system will expand research aiming to unveil new knowledge about memory formation and spatial navigation using novel imaging and computational technologies that can be identified in future experiments as therapeutic targets for the treatment of dementia.

The primary use of this system will be to study a) voltage signals in dendritic compartments and axonal terminals and b) physiological responses from large number of neurons simultaneously. Imaging will be done in 3D custom defined locations which could include various locations along the same dendritic tree or identified somas in from a larger volume. The system will be used for high frequency recording of neural signals from spatially separated and distinct subcellular compartments and for the synchronous stimulation of multiple synaptic inputs terminating on various dendritic regions at a kilohertz rate. The intended use of the system will be to detect single action potentials in trains and subthreshold voltage changes in single trials, with sub millisecond temporal resolution.
 

• This integrated microscope system is suitable for simultaneous fluorescence imaging and photoactivation over the wavelength range of 840 to 1150 nm.
• The imaging system will allow 3D scanning: At a single cell level, recordings from brightly labeled isolated subcompartmental elements such as dendrites and axon terminals opens the way for the optical study of synaptic integration in dendritic compartments and single terminals previously inaccessible to electrodes, which represents a large missing component in our understanding of neuronal function.
• Volumetric scanning: The system will allow scans of local volumes surrounding a target point within tens of microseconds. This is ideal for continuous high-speed high-resolution imaging of electrical activity in deeply located genetically defined neurons during awake behavior. This technology excites reporter molecules over an extended 3D area of membrane, and hence dramatically increases the available signal level and improves signal-to-noise ratios.
• Simultaneous imaging and stimulation can be carried out at 50KHz rate.
• Motorized xyz translation system with control for microscope/stage.
• Ability to visualize neurons for patch-clamp recordings.
• Epifluorescence to increase the collected fluorescence signal in order to increase signal strength and maximum penetration.
• Objectives with high Numerical Aperture (N.A) that allow placement of electrodes (i.e. slim design), one with 10X and the second with 20-25X magnification.
• Integrated software system to operate the microscope system.