Multi-color Three Dimensional Imaging Using Multi-focus Microscopy

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. 

Advantages

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.

Application

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

Related Materials

• Abrahamsson, S., et al, Fast multicolor 3D imaging using aberration-corrected multifocus microscopy. Nature Methods, 10: 60-63 (2013)

Patent Status