Z-microscopy

Patent metadata

Field	Value
Publication number	US-9547163-B2
Application number	US-201314093188-A
Country	US
Kind code	B2
Filing date	Nov 29, 2013
Priority date	Nov 28, 2012
Publication date	Jan 17, 2017
Grant date	Jan 17, 2017

Abstract

We present a method for parallel axial imaging, or z-microscopy, utilizing an array of tilted micro mirrors arranged along the axial direction. Image signals emitted from different axial positions can be orthogonally reflected by the corresponding micro mirrors and spatially separated for parallel detection, essentially converting the more challenging axial imaging to a lateral imaging problem. Each micro mirror also provides optical sectioning capability due to its finite dimension.

First claim

We claim:

1. A method for parallel axial imaging along a depth/axial (z) direction of a microscopy sample, comprising:
directing a test signal to a microscopy sample having an axial dimension along said depth/axial (z) direction;
generating a plurality of imaging signals from said test signal, said plurality of imaging signals emitted from different axial positions along said depth/axial (z) direction;
focusing said imaging signals on a micro mirror array to be reflected;
demultiplexing said plurality of imaging signals through said micro mirror array;
directing said demultiplexed plurality of imaging signals through a plurality of lenses;
collecting said demultiplexed plurality of imaging signals;
measuring said demultiplexed plurality of imaging signals; and
obtaining an axial image or an axial slice image of said microscopy sample from the data;
wherein said test signal is selected from at least one of fluorescence microscopy, confocal microscopy, multiphoton excitation fluorescence microscopy, photoluminescence, light emission, light scattering, harmonic generation microscopy, and Raman microscopy.

2. The method of claim 1 , wherein said micro-mirror array includes a plurality of angled mirrors.

3. The method of claim 2 , wherein said micro-mirror array includes a plurality of mirrors angled at 45 degrees with respect to the depth/axial (z) direction.

4. A device for axial imaging of a sample, comprising:
a means for generation of at least one detecting signal, including at least one excitation lens, wherein said means for generation of said at least one detecting signal is able to be directed to a sample having an axial dimension along a depth/axial (z) direction, generating said at least one detecting signal from different axial positions along said depth/axial (z) direction;
at least one collection lens disposed to focus said at least one detecting signal on a micromirror array to be orthogonally reflected therefrom;
at least one detection lens able to focus said at least one detecting signal from said micromirror array; and
at least one detector array disposed to detect said at least one detecting signal from said detection lens;
wherein said means for generation of a detection signal is selected from at least one of fluorescence microscopy, confocal microscopy, multiphoton excitation fluorescence microscopy, photoluminescence, light emission, light scattering, harmonic generation microscopy, and Raman microscopy.

5. The device of claim 4 , wherein said micromirror array comprises a plurality of mirrors angled at 45 degrees relative to the axial direction of a sample to be tested by the device.

6. The device of claim 4 , wherein a Fresnel lens is used to achieve an excitation beam with chromatically extended depth of focus so that a plurality of image signals are generated along the axial direction.

7. The device of claim 4 , wherein said micromirrors have an aspect ratio of height to width of at least 10:1.

8. A method for axial slice light sheet imaging of a specimen, comprising:
subjecting a specimen to a line of excitation beam to generate image signals, said specimen having an axial dimension along a depth/axial (z) direction, said image signals being generated from different axial positions along said depth/axial (z) direction;
imaging the signals onto a plurality of micro mirrors arrayed at an angle relative to the axial direction;
de-multiplexing said signals;
detecting the de-multiplexed signals; and
outputting the detected de-multiplexed signals as a slice plane image of the specimen;
wherein the line of excitation beam is prepared by at least one of scanning a beam with a chromatically extended depth of focus, scanning a Bessel beam, focusing a beam with a cylindrical lens, scanning a beam with a chromatically extended depth of focus produced with a Fresnel lens, and scanning an Airy beam.

9. The method of claim 8 , wherein said micro mirrors are arrayed at an angle of 45 degrees relative to said depth/axial (z) direction.

10. The method of claim 2 , wherein said mirrors are angled at about 45 degrees.

11. The method of claim 1 , further comprising excitation over an extended axial range through chromatic aberration.

12. The method of claim 8 , further comprising introducing chromatic aberration via excitation over a chromatically extended axial range with a Fresnel lens.

13. The method of claim 8 , wherein said image signals are fluorescent signals and said fluorescent signals are excited and demultiplexed before being detected.

14. The method of claim 8 , wherein said at least one detecting signal is excited and demultiplexed within a chromatically extended axial range.

15. The method of claim 13 , wherein said fluorescent signal is excited and demultiplexed within a chromatically extended axial range.

16. The method of claim 8 , wherein said slice plane image of said specimen is a whole two dimensional axial slice performed by taking a single shot with a 2D imager.

Assignees

• Liu Zhiwen
• Yang Chuan
• Shi Kebin
• Zhou Mingda
• Zheng Siyang
• Yin Shizhuo
• Penn State Res Found

Inventors

• Liu Zhiwen
• Yang Chuan
• Shi Kebin
• Zhou Mingda
• Zheng Siyang
• Yin Shizhuo

Classifications

• G02B21/008 Primary

  Details of detection or image processing, including general computer control · CPC title

• G02B21/006

  focusing arrangements; selection of the plane to be imaged · CPC title