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High Speed Scanning Image

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Imaging Instrumentation
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High Speed Scanning Image

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Volume imaging of thick samples at an ultra high-speed

Laser scanning microscopy (LSM) is an essential tool for imaging biological specimens, and two-photon LSM is the standard for imaging in vivo or in other highly scattering media. However, imaging speeds have traditionally been limited by the rate at which the laser beam can be rastered across a sample. Recently, random access scanning has been used to vastly improve the speed of volumetric imaging by scanning only predefined areas of interest within the 3D space; data acquisition rates are roughly the same. Nevertheless, there remains a need to image continuous volumetric spaces at fast scanning rates so that researchers can adequately study dynamic and unpredictable events.

The use of phase-locked ultrasound lenses has recently been incorporated by inventors at Howard Hughes Medical Institute’s Janelia Research Campus to overcome such scanning speed limitations. This innovation utilizes an ultrasound lens to modulate the axial position of the focal volume, allowing a sampling rate of 80 MHz. This rate matches the repetition rate of the laser source; axial line scans can be acquired in ~1 µs and large 3D scans at 1-10 Hz or higher depending on the volume size. Unlike systems that rely on acousto-optical deflectors to rapidly move the excitation beam, the ultrasound lens introduces almost no optical losses. Moreover, the ultrasound lens can be incorporated at the front of the optical train and readily coupled to existing LSM instruments. The end result is volumetric imaging rates that cannot be approached by current methods. The system’s capabilities have been demonstrated by acquiring images of transient neuronal activity as well as changes in cell morphology that could not be adequately measured using any other described technique.

Advantages:

  • Volumetric two-photon imaging in thick tissues at unprecedented speeds
  • Almost zero attenuation of the excitation source
  • 3D images over continuous space ensure that events are not missed
  • Add-on to existing laser scanning microscopes

Applications:

  • Research studies in neuroscience, immunology, and stem cell research that demands observation of 3-dimensional transient events
  • Fast 3D calcium imaging
  • Recording in vivo dynamics such as cell movements and morphology changes
  • Blood flow imaging

Patent Status:

U.S. patent ​9,720,218

Opportunity:

Free for Non-Profit Research and available for Commercial License. Please contact Innovation Management at innovation@janelia.hhmi.org.

For inquiries, please reference:

Janelia 2013-034

 

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Contact

Michael Perham
Head of Innovation Management
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