Multi-color Three Dimensional Imaging Using Multi-focus Microscopy

Tech ID: 23832 / UC Case 2012-220-0

Invention Novelty

This technology identifies an advanced imaging technique of biomicroscopy using an innovative type of a wide-field multi-focus microscope to enable fast, high-resolution 3D imaging. 


This technology provides the following advantages: 


  • Powerful tool for fast and sensitive 3D fluorescence microscopy of live samples
  • Simultaneous acquisition of all focal planes on a single camera
  • Light efficiency of the imaging path is better than 50 percent
  • Current design allows imaging of a volume of 33x33x18 microns at 31Hz 


Technology Description

Today, many three-dimensional (3D) biological specimens and processes are studied in two-dimensions (2D) due to the slow speed of the light microscope in recording a 3D image. This technology advances imaging techniques of biomicroscopy using an innovative type of a wide-field Multi-Focus microscope to enable fast, high-resolution 3D imaging. The system includes a novel grating design and chromatic correction scheme that is appended to the camera port of a high-resolution epifluorescence microscope to produce an instant focal stack of high-resolution 2D images simultaneously displayed on a single camera. The 3D microscope is designed to minimize photobleaching and phototoxicity while enabling high-speed imaging of weak fluorescent samples such as single fluorophores and to acquire multiple focal planes without aberrations to avoid loss in resolution and contrast.  

The concept of Multi-Focus imaging uses optical manipulation to produce an entire focal series instantaneously. The Multi-Focus system developed is based on a phase-only diffractive grating placed in the Fourier plane in the emission path. The grating pattern is specially designed to split up the emission light from the sample into a set of simultaneous images and to focus shift diffraction orders. Each diffraction order forms a separate image of the sample, and the resulting series of images is recorded as a 2D array on a single, large-chip electron multiplying charge couple device camera.


  • Basic life science research 
  • Fluorescence biomicroscopy 
  • Single particle tracking in live cells 
  • Neuronal imaging 
  • Development imaging of small organisms

Related Materials

Patent Status

Country Type Number Dated Case
United States Of America Issued Patent 9,477,091 10/25/2016 2012-220
European Patent Office Published Application 2802861 11/19/2014 2012-220

Additional Patent Pending


Other Information


Multi-color, Multi-Focus, Microscopy, 3D, biomicroscopy, Fluorescence, Single Particle

Categorized As


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