Columbia Technology Ventures

Holographic 3D imaging and photostimulation for enhanced functional assays of neuroactive compounds

This technology is a microscope platform that combines holographic beam multiplexing with the power of two-photon optogenetics and uncaging of photosensitive compounds for high-resolution imaging and mapping of neural circuits and functional assaying of the effect of pharmacological compounds on individual neurons.

Unmet Need: High-resolution microscope for mapping neuronal circuitry in 3D

Detecting activity of individual neurons and the spatiotemporal pattern in which they fire is an important step to understanding brain function. The modern laser scanning microscope (LSM) is a powerful tool that can be used to effectively image and optically manipulate neurons. A prominent disadvantage for such a system, however, is the generally poor temporal resolution which is limited by serial scanning rate. There are several ways to improve temporal resolution in LSM systems. One possible approach is to split the excitation light into multiple beamlets and scan different locations simultaneously. Alternatively, spinning-disk confocal microscopy or semitransparent mirror beam-splitters and galvanometer scanners can be used. However, these strategies limit the energy that is effectively available in the laser and result in a lower signal to noise ratio for the acquired image.

The Technology: Enhanced two-photon microscopy for high-resolution imaging and mapping of neuronal circuits

This technology describes an imaging technique for increasing temporal resolution in microscope systems using two-photon holographic imaging and stimulation. Specifically, the microscope is adapted with a diffractive spatial light modulator (SLM), a holographic device capable of modifying the phase profile of a laser wavefront to build a 3D light pattern, placed in the beam path of a two-photon microscope to generate multiple beamlets that can be independently and dynamically controlled. As a result, the temporal resolution is only limited by the detection rate instead of the scanning rate, which greatly improves the function of the microscope system. This technology enables simultaneous 3D imaging of neurons, eliminates wasteful wall time (repeat scans of areas that are not actively changing), and saves on total illumination power, all while delivering images at greater speeds. Furthermore, this device can stimulate individual neurons in brain slices in any spatio-temporal pattern in 3D, using two-photon optogenetics or uncaging of photo-sensitive compounds, for real time manipulation and mapping of neuronal circuits.

This technology has been demonstrated in vivo on several layers of the primary visual cortex of a living mouse and can provide a selective assay to test the effect of neuroactive compounds in neural circuits of living animals.

Applications:

  • Laser microscopes
  • Cell imaging
  • Functional imaging of neural circuits in vivo
  • Functional assay for studying neuroactive compounds
  • Clinical tool for visualizing brain function in normal and altered mental or neurological states

Advantages:

  • Eliminates wasteful functioning
  • Adaptive optics
  • High spatiotemporal 3D imaging
  • Non-invasive imaging technique
  • Reduced exposure to harmful radiation
  • Faster imaging speeds than conventional two-photon microscopy methods

Lead Inventor:

Rafael Yuste, Ph.D.

Patent Information:

Patent Issued Patent Status

Related Publications:

Tech Ventures Reference: