NU 2018-070

Inventors

Oliver Cossairt*

Kuan He

Aggelos K. Katsaggelos

Norbert Sherer

Mark Hereld

Short Description

3D fluorescent imaging with high temporal and spatial resolution

Background

A key hurdle in modern 3D fluorescent imaging is achieving both high temporal and spatial resolution across large volumes. Light microscopes use focal scanning to acquire 3D images, which is temporally slow. Spatial resolution, and in particular axial resolution, is limited due to the mechanical constraints of objective lenses. To overcome focal scanning and achieve high temporal resolution, multifocus microscopy (MFM) captures images at many planes at a time. However, MFM suffers from poor axial resolution. Image interference microscopy (I2M) achieves high axial resolution, but still scans the Z plane, making it temporally slow. This new technology, interferometric multifocus microscopy (iMFM), overcomes both temporal and axial limitations.

Abstract

Northwestern researchers have developed a method of microscopy that provides snapshot super-resolution 3D imaging in multiple planes, called interferometric multifocus microscopy (iMFM). In optical microscopy for live imaging applications, there are trade-offs between temporal and spatial resolution, especially in regard to axial resolution. This trade-off stems from the limitations of light and the inherently slow nature of mechanical scanning. Using an interferometric approach combined with multifocus microscopy (MFM), iMFM achieves isotropic 3D with localization accuracy on the nanoscale, with superior axial resolution to MFM. The new approach used by iMFM therefore overcomes the tradeoff between temporal and spatial resolution. This technology has the potential to benefit 3D dynamic event imaging by simultaneously providing high temporal and spatial resolution.

Applications

• Single molecule/cell tracking
• High temporal and spatial resolution 3D microscopy imaging of extended objects, such as bacteria

Advantages

• Simultaneous solution to microscopy’s temporal and spatial resolution issues
• Isotropic 3D nanoscale localization accuracy
• Easier image acquisition than other comparable systems

Publications

He K, Wang  Z, Huang X, Wang X, Yoo S, Ruiz P, Gdor I, Selewa A, Ferrier NJ, Scherer N, Hereld M, Katsaggelos AK, and Cossairt O (2018) Computational multifocal microscopy. Biomedical Optics Express 9:6477-6496.

IP Status

A provisional application has been filed.

The iMFM system is comprised of of two opposing objectives, a beam splitter (BS), and a detector. The two multi-focus gratings (MFGs) placed in the Fourier planes of each objective are capable of producing multifocal interferometry detection on the BS, which is imaged via lenses 3 & 4 (L3 & L4) onto the detector in a single shot.