Extended depth of focus microscopy

• US 20040228520A1
• Filed: 02/13/2004
• Published: 11/18/2004
• Est. Priority Date: 02/18/2003
• Status: Active Grant

• Current Assignee: Oklahoma Medical Research Foundation
• Original Assignee: Oklahoma Medical Research Foundation
• Inventors: Dresser, Michael E., Conchello, Jose-Angel
• Granted Patent: US 7,444,014 B2
• Time in Patent Office: Days
• US Class Current: 382/154
• CPC Class Codes: G06T 2200/04 involving 3D image data
• CPC Class Codes: G06T 2207/10056 Microscopic image
• CPC Class Codes: G06T 2207/10064 Fluorescence image
• CPC Class Codes: G06T 2207/10148 Varying focus
• CPC Class Codes: G06T 5/003 Deblurring; Sharpening vibration or motion blur correction for cameras comprising an electronic image sensor H04N23/682
• CPC Class Codes: G06T 5/73 Deblurring; Sharpening
• CPC Class Codes: G06T 7/50 Depth or shape recovery
• CPC Class Codes: G06V 20/693 Acquisition
• CPC Class Codes: G06V 2201/12 Acquisition of 3D measurements of objects

Abstract

Imaging techniques. The focus of an imaging device is varied while acquiring an image of an object. The resulting blurred image is deconvolved to generate, in one embodiment, a two-dimensional projection image of three dimensions of the object.

Claims

1. An imaging method comprising:

displacing an imaging device in one dimension while acquiring an image of an object, thereby blurring the image; and

deconvolving the blurred image to generate a multidimensional representation of the object.

2. An imaging method comprising:

varying the focus of an imaging device while acquiring an image of an object, thereby blurring the image; and

deconvolving the blurred image to generate a representation of the object.

3. The method of claim 2, the representation comprising a two dimensional projection image of three dimensions of the object.

4. The method of claim 2, the imaging device comprising a fluorescence imaging device.

5. The method of claim 2, varying the focus occurring while a shutter of the imaging device is open.

6. The method of claim 2, varying the focus comprising varying an input voltage to a piezoelectric focusing mechanism of the imaging device.

7. The method of claim 2, varying the focus comprising applying signals to a piezoelectric focusing mechanism of the imaging device to generate oscillatory movement of the focusing mechanism.

8. The method of claim 2, varying the focus comprising launching a velocity-controlled focus change using the a stand-based focusing mechanism.

9. The method of claim 2, acquiring the image being accomplished in two or more stages.

10. An imaging method comprising:

(a) collecting an acquired image of an object using an imaging device;

(b) varying the focus of the imaging device while collecting the acquired image, thereby blurring the acquired image;

(c) determining a point spread function (PSF) associated with the imaging device;

(d) determining an optical transfer function (OTF) using the PSF;

(e) determining an object estimate;

(f) convolving the object estimate with the PSF, using the OTF, to generate an estimated image;

(g) comparing the estimated image with the acquired image to obtain a ratio;

(h) convolving the ratio with a mirror image of the PSF, using a complex conjugate of the OTF, to form a convolved ratio;

(i) multiplying the object estimate with the convolved ratio to form an updated object estimate; and

(j) repeating steps (f) through (i) one or more times to generate a two dimensional projection image of three dimensions of the object from the updated object estimate.

11. The method of claim 10, the imaging device comprising a fluorescence imaging device.

12. The method of claim 10, the imaging device comprising a photosensitive camera chip.

13. The method of claim 10, collecting the acquired image comprising stopping a continual clearing of the imaging device.

14. The method of claim 10, collecting the acquired image comprising opening a shutter of the imaging device.

15. The method of claim 14, varying the focus occurring while a shutter of the imaging device is open.

16. The method of claim 10, varying the focus comprising varying an input voltage to a piezoelectric focusing mechanism of the imaging device.

17. The method of claim 10, varying the focus comprising applying signals to a piezoelectric focusing mechanism of the imaging device to generate oscillatory movement of the focusing mechanism.

18. The method of claim 10, varying the focus comprising launching a velocity-controlled focus change using the a stand-based focusing mechanism.

19. The method of claim 10, acquiring the image being accomplished in two or more stages.

20. An imaging system, comprising:

an imaging device configured to vary its focus while acquiring an image of an object; and

a processor in operative relation with the imaging device and configured to execute machine-readable instructions for deconvolving a resulting blurred image to generate a representation of the object.

21. The system of claim 20, the representation comprising a two dimensional projection image of three dimensions of the object.

22. The system of claim 20, the imaging device comprising a fluorescence imaging device.

23. The system of claim 20, the instructions comprising instructions for:

(a) determining a point spread function (PSF) associated with the imaging device;

(b) determining an optical transfer function (OTF) using the PSF;

(c) determining an object estimate;

(d) convolving the object estimate with the PSF, using the OTF, to generate an estimated image;

(e) comparing the estimated image with the acquired image to obtain a ratio;

(f) convolving the ratio with a mirror image of the PSF, using a complex conjugate of the OTF, to form a convolved ratio;

(g) multiplying the object estimate with the convolved ratio to form an updated object estimate; and

(g) repeating steps (d) through (g) one or more times.

24. A retrofit kit for converting an imaging system, comprising:

means for allowing an imaging device to vary its focus while acquiring an image of an object; and

means for deconvolving a blurred image to generate a two dimensional projection image of three dimensions of the object.