JPWO2020205952A5

JPWO2020205952A5 -

Info

Publication number: JPWO2020205952A5
Application number: JP2021559237A
Authority: JP
Publication date: 2023-03-03

Claims

A method of imaging a sample, comprising:
illuminating the plurality of points of the sample with a first converging fringe pattern produced at each of the plurality of points by interference of light beams emitted through a microscope objective;
For said first converging fringe pattern, a first discrete illumination area of said microscope objective lens has a first axis of rotation and said light beams are phased with respect to each other by a first phase. shifted and
The method further comprises illuminating the plurality of points with at least one different converging fringe pattern, wherein the light beams are phase shifted with respect to each other by different phases relative to the first phase, and ( i) the same separate illumination areas of the microscope objective have the same axis of rotation that is not angled with respect to the first axis of rotation; or (ii) different illumination areas of the microscope objective. separate illumination areas having different axes of rotation angled with respect to the first axis of rotation;
The method, wherein one or both of the discrete illumination areas are non-circular.

further comprising acquiring illumination data for each of a plurality of raw images corresponding to each of said first convergent fringe pattern and said at least one different convergent fringe pattern from a single element detector or spectrograph. Item 1. The method according to item 1.

further comprising reconstructing a super-resolution composite image from the illumination data for each of the plurality of raw images according to the formula:

wherein I _syn is the composite image, n _images is the number of each of the plurality of raw images,
for each of the first converging fringe pattern and the at least one different converging fringe pattern;
i is an index corresponding to the rotation axis angle,
j is an index corresponding to the phase shift of the light beams with respect to each other;
_Irawi,j is the illumination data for each of the plurality of raw images, with index i corresponding to the axis of rotation angle and index j corresponding to the phase shift;
A is a coefficient representing the contrast of the converging fringe pattern,
3. The method of claim 2, wherein phase is the phase shift.

A method according to any one of claims 1 to 3, wherein said discrete illumination area comprises a maximum inscribed circle having a center substantially collinear with the center of the rear aperture of said microscope objective. .

5. The method of claim 4, wherein each of said separate illumination areas further comprises an illumination enhancement area outside said largest inscribed circle.

6. The method of claim 5, wherein said illumination enhancement area represents at least about 50% of said discrete illumination area.

6. The method of claim 5, wherein the illumination area is shaped as a slit coinciding with the opposite portion of the rear circular aperture of the microscope objective.

A method according to any preceding claim, wherein for said at least one different converging fringe pattern said light beams are modulated in phase using a programmable spatial light modulator.

9. A method according to any one of the preceding claims, wherein for neither the first nor the at least one different converging fringe pattern the distinct illumination areas are phase shifted with a cosine of zero. the method of.

Claims 1 to 9, wherein for neither said first nor said at least one different converging fringe pattern said separate illumination regions are phase shifted by a cosine with an absolute value of less than 0.5. The method according to any one of .

The at least one different convergent fringe pattern is a second convergent fringe pattern in which the light beam is shifted by a second phase different from the first phase, and the same distinct illumination area is the A method according to any one of claims 1 to 10, wherein the second separate illumination area has a second axis of rotation that is not angled with respect to the first axis of rotation.

The at least one different converging fringe pattern is
wherein the light beam is shifted by a third phase that is substantially the same as the first phase, and a third separate illumination area is aligned with the first axis of rotation and the second axis of rotation; 12. The method of claim 11, further comprising a third converging fringe pattern having a third axis of rotation that is angled relative to.

The at least one different converging fringe pattern is
the light beam is shifted by a fourth phase that is substantially the same as the second phase, and a fourth discrete illumination area is not angled with respect to the third axis of rotation 13. The method of claim 12, further comprising a fourth axis of rotation.

A method according to any one of claims 11 to 13, wherein said second phase differs from said first phase by π.

A method according to any one of claims 11 to 14, wherein said first phase is 0 and said second phase is π.

The at least one different converging fringe pattern is
wherein the light beam is shifted by a fifth phase substantially identical to the first phase and the third phase, and a fifth separate illumination area is shifted from the first, second, third, and a fifth converging fringe pattern having a fifth axis of rotation angled with respect to the fourth axis of rotation;
the light beam is shifted by a sixth phase substantially identical to the second phase and the fourth phase, and a sixth separate illumination area is angled with respect to the fifth; 14. The method of claim 13, comprising a sixth converging fringe pattern having a non-zero sixth axis of rotation.

17. The method of claim 16, wherein said first, third and fifth phases are 0 and said second, fourth and sixth phases are [pi].

The third axis of rotation has an angle of about π/6 to about π/2 with respect to the first axis of rotation, and the fifth axis of rotation has an angle of about π/6 with respect to the first axis of rotation. 17. The method of claim 16, having an angle of [pi]/2 to about 5[pi]/6.

A method of imaging a sample, comprising:
acquiring illumination data for each of a plurality of raw images, each of said raw images corresponding to a given one of a set of converging fringe patterns, each of said converging fringe patterns corresponding to a microscope objective; produced by the interference of a pair of light beams emitted through a lens to illuminate a plurality of points on the sample;
The set of converging fringe patterns includes an X and Y component that is a combination of (i) the X phase at which the light beam is shifted and (ii) the Y angle at which the axis of rotation of the light beam is oriented. , the set is less than or equal to two different values of X;
The method of claim 1, wherein the light beams of the pair of light beams illuminate a non-circular illumination area of the microscope objective lens for at least one of the converging fringe patterns.

Apparatus for performing Raman spectroscopy or fluorescence microscopy, comprising a programmable spatial light modulator (SLM) for modulating the phase of the light beam, and comprising a programmable spatial light modulator (SLM) for modulating the phase of the light beam. Apparatus configured to perform the method of claim 1.

21. The apparatus of claim 20, wherein the SLM displays a phase grating that passes the light beam through the non-circular discrete illumination areas of the microscope objective.

22. Apparatus according to claim 20 or 21, arranged to perform a Gaussian blur to smooth the non-circular discrete illumination areas.