Systems and methods for image processing in optical microscopes

1. A method for processing an image in optical microscopy, the method comprising:

acquiring a raw image with a microscope, wherein the raw image has an asymmetric resolution that is different in an X direction than in an orthogonal Y direction;

correcting the raw image using at least one of a X-direction point spread function and a Y-direction point spread function in order to generate an asymmetrically deconvolved image; and

displaying the asymmetrically deconvolved image on a display device,

wherein correcting the raw image further comprises:

determining the X-direction point spread function and applying the X-direction point spread function to deconvolve the X direction; and

determining the Y-direction point spread function and applying the Y-direction point spread function to deconvolve the Y direction,

wherein the X-direction and Y-direction are deconvolved independently.

2. The method of claim 1 , wherein the raw image has an elliptical point spread function with a confocal point spread function in the Y-direction and a wide field point spread function in the X-direction.

3. The method of claim 2 , further comprising:

determining the elliptical point spread function based at least in part on the wide field point spread function in the X-direction.

4. The method of claim 2 , further comprising:

deconvolving the X-direction using the wide field point spread function; and

deconvolving the Y-direction using the confocal point spread function.

5. The method of claim 4 , wherein the confocal point spread function is a square root of the wide field point spread function.

6. The method of claim 1 , wherein a photon collection efficiency of the microscope is at least 10% of an equivalent wide-field microscope.

7. The method of claim 1 , wherein the microscope is a line-scan confocal microscope, live-cell confocal microscope, “spinning disk” microscope, fluorescent microscope, deconvolution microscope, or a point-scan confocal microscope.

8. The method of claim 1 , wherein the raw image is a monochromatic fluorescent image.

9. The method of claim 1 , wherein deconvolving the X-direction or Y-direction comprises Baysian deconvolution, Richardson-Lucy deconvolution, Wiener deconvolution, Fourier deconvolution, wavelets, or other image processing methods that compute deconvolution.

10. The method of claim 1 , wherein correcting the raw image further comprises:

deriving the Y-direction point spread function using the X-direction point spread function; and

applying the derived Y-direction point spread function to deconvolve the Y-direction.

11. The method of claim 1 , wherein the X-direction point spread function is different from the Y-direction point spread function.

12. A system for processing an image in an optical microscopy, the system comprising:

(a) an input device configured to provide image data from an optical detector; and

(b) one or more processors coupled to the input device and configured for correcting the image using at least one of a X-direction point spread function and a Y-direction point spread function to generate an asymmetrically deconvolved image,

wherein correcting the image further comprises:

determining the X-direction point spread function and applying the X-direction point spread function to deconvolve the X direction; and

determining the Y-direction point spread function and applying the Y-direction point spread function to deconvolve the Y direction,

wherein the X-direction and Y-direction are deconvolved independently.

13. The system of claim 12 , wherein the one or more processors comprises one or more of a central processing units, a graphics processing unit, and a floating point unit.

14. The system of claim 12 , wherein the image comprises a monochromatic fluorescent image with an asymmetric resolution.

15. The system of claim 12 , wherein the image comprises an elliptical point spread function with a confocal point spread function in the Y direction and a wide field point spread function in the X direction.

16. The system of claim 15 , wherein the one or more processors is further configured to:

determine the elliptical point spread function based at least in part on the wide field point spread function in the X direction.

17. The system of claim 15 , wherein the one or more processors is further configured to:

deconvolve the X direction using the wide field point spread function; and

deconvolve the Y direction using the confocal point spread function.

18. The system of claim 15 , wherein the confocal point spread function is a square root of the wide field point spread function.

19. The system of claim 12 , wherein the input device comprises a line-scan confocal microscope, a live-cell confocal microscope, a “spinning disk” microscope, a fluorescent microscope, a deconvolution microscope, or a point-scan confocal microscope.

20. The system of claim 19 , wherein a photon collection efficiency of the microscope is at least 10% of an equivalent wide-field microscope.

21. The system of claim 12 , wherein deconvolving the X or Y direction comprises Baysian deconvolution, Richardson-Lucy deconvolution, Wiener deconvolution, Fourier deconvolution, wavelets, or other image processing methods that compute deconvolution.

22. An image processing system for processing a monochromatic image from a CMOS camera-based line-scan confocal fluorescent microscope, the system comprising:

(a) one or more CMOS cameras capable of generating the monochromatic image; and

(b) a system for processing the monochromatic image, the system comprising:

(1) an input device configured to provide image data from the CMOS camera; and

(2) one or more processors coupled to the input device and configured for:

correcting the monochromatic image using at least one of a X-direction point spread function and a Y-direction point spread function to generate an asymmetrically deconvolved image,

wherein correcting the monochromatic image further comprises:

determining the X-direction point spread function and applying the X-direction point spread function to deconvolve the X direction; and

determining the Y-direction point spread function and applying the Y-direction point spread function to deconvolve the Y direction,

wherein the X-direction and Y-direction are deconvolved independently.