Innovation and Partnerships Office

Axial and Arbitrary Plane Optical Microscope 2013-153

An optical microscope for fast, high contrast 2-D and 3-D imaging of structures beneath the surface of large biological samples, such as the cerebral cortex of live mammal brains. Compatible with conventional wide-field microscopes, the Berkeley Lab technology offers a ready way to improve the imaging capabilities of conventional optical microscopes.

APPLICATIONS OF TECHNOLOGY:

  • Commercial manufacturing of optical microscopes
  • Imaging beneath the surface of large, live biological samples for
    • Diagnosis of disease such as neurological disorders
    • Medical research
    • Mechanical studies of cells and cellular processes

ADVANTAGES:

  • Fast, high contrast 2-D imaging of an arbitrary desired plane does not require scanning or computation
  • High contrast, convenient 3-D imaging
  • Compatible with commercially available wide-field optical microscopes
  • Images biological samples of any size
  • High signal-to-noise ratio imaging with reduced photo bleaching

ABSTRACT:

Despite the popularity of conventional optical microscopes such as confocal microscopes and selective-plane illumination microscopes (SPIMs), there are no commercially available optical microscopes that can quickly produce high contrast 2-D images or 3-D images of structures beneath the surface of large, live samples such as the cerebral cortex.

Researchers at Berkeley Lab have developed an axial plane optical microscope (APOM) that directly images a sample’s cross-section parallel to the optical axis of an objective lens or an arbitrary cross-section of interest without scanning, making it inherently fast. In addition, it enables convenient, fast, and high contrast 3-D imaging of large biological tissues by scanning along one direction only rather than point-by-point.

The invention is compatible with conventional wide-field microscopes and can be used to extend the imaging capacity of conventional 2-D wide-field microscopes from a fixed focal plane to arbitrary cross-sections of interest as well as the 3-D imaging of large, live biological samples. Compared to techniques such as laser-scanning confocal microscopy, the Berkeley Lab technology offers faster imaging, higher signal-to-noise ratio, reduced overall photo bleaching and more convenient imaging, as it does not require special sample preparation.

Conventional wide field microscopy is not ideal for imaging large biological tissues or for single-molecule fluorescence imaging inside a living cell. It is limited to imaging a sample’s cross-sections within the objective’s thin focal (lateral) plane because it requires two bulky objective lenses aligned very close to each other, limiting the sample shape and/or the thickness of illumination light sheet.

In contrast, the Berkeley Lab APOM technology uses one objective lens near the sample for both illumination and image collection, and another objective lens far away from the sample to form an intermediate optical 3-D image. The axial plane image of the sample is formed at the image plane of the remote objective lens and collected by a camera or observed by the naked eye.

This versatile instrument can be turned into an arbitrary plane optical microscope simply by rotating the tilted mirror around the optical axis to image a general cross-section of a sample that is not parallel or perpendicular to the optical axis.

DEVELOPMENT STAGE:  Proven principle

STATUS:  Published U. S. Patent Application 14/591,760.  Available for licensing or collaborative research.

FOR MORE INFORMATION:

Jeongmin Kim, Tongcang Li, Yuan Wang, and Xiang Zhang, “Vectorial point spread function and optical transfer function in oblique plane imaging,” Optics Express, 22, 11140-11151 (2014).

SEE THESE OTHER BERKELEY LAB TECHNOLOGIES IN THIS FIELD:

Integrated Profiling of 3D Cell Culture Models and 3D Microscopy, 2013-177

The Campanile Tip for Optimized Nanoscale Imaging, IB-3314

Stable Imaging Arrangement and Fluorescent Microscope, JIB-2619

REFERENCE NUMBER: 2013-153