Microscope and method for computational microscopic layer separation
A microscope for computational microscopic layer separation may include an imaging device that includes a lens and an image sensor, an illumination system for illuminating a sample, and an actuator to adjust an axial position of a focal plane with respect to the sample. The microscope may also include a processor operatively coupled to the imaging device and the illumination system. The processor may be configured to measure, using the image sensor and the illumination system, optical aberrations of the imaging device at the axial position, and determine whether to adjust the focal plane with respect to the sample in response to the one or more optical aberrations. Various other systems and methods are also disclosed.
1. A microscope comprising:
at least one imaging device comprising lens and an image sensor;
an illumination system for illuminating a sample;
an actuator to adjust an axial position of a focal plane with respect to the sample; and
a processor operatively coupled to the at least one imaging device and the illumination system, the processor configured to:
measure, using the image sensor and the illumination system, one or more optical aberrations of the imaging device at the axial position; and
determine whether to adjust the focal plane with respect to the sample in response to the one or more optical aberrations, wherein the processor is configured to determine whether the one or more optical aberrations corresponds to a presence of a transparent material covering the sample between the lens and the focal plane;
wherein the at least one imaging device is configured to image an object with decreased aberrations with the focal plane of the imaging device located beneath an upper surface of the transparent material as compared to the focal plane located on or above the upper surface.
2. The microscope of claim 1 , wherein the processor is configured to determine whether the focal plane is located on one or more of on a surface of the transparent material covering the sample, beneath the surface of the transparent material covering the sample or within the sample in response to an amount of the one or more aberrations.
3. The microscope of claim 2 , wherein the imaging device is configured to image the sample with the transparent material covering the sample with a decreased amount of aberrations as compared to the transparent material not covering the sample and wherein the processor is configured with instructions to adjust the focal plane toward the sample in order to decrease the one or more aberrations.
4. The microscope of claim 2 , wherein the processor is configured to determine whether the focal plane is located on dirt on a surface of the transparent material covering the sample or on the sample beneath the surface of the transparent material in response to the amount of the one or more aberrations and a structure of an image captured with the at least one imaging device at the axial position.
5. The microscope of claim 4 , wherein the sample comprises a sparse sample.
6. The microscope of claim 2 , wherein the transparent material covering the sample comprises one or more of a solid coverslip or a liquid coverslip.
7. The microscope of claim 2 , wherein the transparent material covering the sample comprises an index of refraction of at least 1.4.
8. The microscope of claim 1 , wherein the illumination system is configured to illuminate the sample at a plurality of illumination angles.
9. The microscope of claim 8 , wherein the processor is configured to:
capture, using the image sensor, image data from a plurality of images at the axial position using the plurality of illumination angles;
calculate an amount of shift in the image data between the plurality of images; and
determine, using the calculated amount of shift, a distance between the sample and the focal plane.
10. The microscope of claim 9 , wherein the processor is configured to determine the amount of the one or more aberrations in response to the calculated amount of shift.
11. The microscope of claim 9 , wherein the processor is configured to illuminate the sample from the plurality of illumination angles simultaneously with simultaneous activation of a plurality of light sources.
12. The microscope of claim 11 , wherein the processor is configured to illuminate the sample from the plurality of illumination angles sequentially with sequential activation of the plurality of light sources.
13. The microscope of claim 1 , wherein the amount of the one or more optical aberrations is compared to a predetermined value of the one or more optical aberrations and optionally wherein the axial position is adjusted in response to the amount being greater than the predetermined value or not adjusted in response to the amount being less than the predetermined value.
14. The microscope of claim 1 , wherein the processor is configured with instructions to determine whether an object located at the focal plane is in focus in response to an image shift from images captured with the image sensor and to adjust the axial position of the focal plane in response to the object being in focus and an amount of the one or more aberrations above a threshold amount.
15. The microscope of claim 1 , wherein the one or more optical aberrations comprises a spherical aberration.
16. The microscope of claim 15 , wherein the one or more optical aberrations comprises one or more of coma or astigmatism.
17. The microscope of claim 1 , wherein the one or more optical aberrations does not comprise defocus.
18. The microscope of claim 1 , wherein the one or more optical aberrations comprise two or more optical aberrations and wherein the two or more optical aberrations include defocus.
19. The microscope of claim 1 , wherein the focal plane comprises an axial location within a depth of field of the at least one imaging device.
20. A method comprising:
illuminating a sample with an illumination system;
imaging the sample with at least one imaging device comprising lens and an image sensor;
adjusting, with an actuator, an axial position of a focal plane with respect to the sample; and
measuring, using the image sensor and the illumination system, one or more optical aberrations of the imaging device at the axial position; and
determining, with a processor, whether to adjust the focal plane with respect to the sample in response to the one or more optical aberrations, wherein the processor determines whether the one or more optical aberrations corresponds to a presence of a transparent material covering the sample between the lens and the focal plane, wherein the at least one imaging device is configured to image an object with decreased aberrations with the focal plane of the imaging device located beneath an upper surface of the transparent material as compared to the focal plane located on or above the upper surface.