Imaging apparatus and image generating method

• US 9,100,598 B2
• Filed: 10/20/2010
• Issued: 08/04/2015
• Est. Priority Date: 10/29/2009
• Status: Active Grant

• Current Assignee: Evident Corporation
• Original Assignee: Olympus Corporation
• Inventors: Aizaki, Shinichiro, Kaminaga, Shigeto
• Primary Examiner(s): Le, Thao X
• Assistant Examiner(s): LE, LONG HUYNH
• Application Number:
  US12/908,270
• Publication Number:
  US 20110102578A1
• Time in Patent Office: 1,749 Days
• Field of Search: 348/135, 348/222.1, 382/275, 600/476
• US Class Current: 1/1
• CPC Class Codes: G02B 21/16 adapted for ultraviolet illumination ; Fluorescence microscopes G02B21/0076 takes precedence
• CPC Class Codes: G02B 21/365 Control or image processing arrangements for digital or video microscopes G02B21/361, G02B21/362 take precedence
• CPC Class Codes: H04N 23/70 Circuitry for compensating brightness variation in the scene
• CPC Class Codes: H04N 23/71 Circuitry for evaluating the brightness variation
• CPC Class Codes: H04N 23/741 by increasing the dynamic range of the image compared to the dynamic range of the electronic image sensors
• CPC Class Codes: H04N 25/48 Increasing resolution by shifting the sensor relative to the scene

Abstract

An imaging apparatus includes: an imaging unit for capturing an image of a fluorescent sample; a pixel shift unit for changing the relative position of the imaging unit for the image; an image correction unit for correcting the gray-scale levels of plural pieces of image data using the histograms of the image data acquired by the imaging unit at different relative positions; and an image combination unit for combining the plural pieces of image data corrected by the image correction unit.

Claims

1. An imaging apparatus, comprising:

an imaging unit configured to capture a plurality of images of a fluorescent sample;

a pixel shift unit configured to change a relative position of the imaging unit for each of the plurality of images;

a processor; and

a memory storing computer readable instructions that, when executed by the processor, implement;

an image correction unit correcting, by setting as a reference image one of the plurality of images acquired by the imaging unit at one of the different relative positions, a gray-scale level of at least one correction target image selected from the plurality of images other than the reference image so that each histogram of each respective correction target image other than the reference image matches a histogram of the reference image; and

an image combination unit combining the reference image and each correction target image corrected by the image correction unit so as to generate image data with a higher resolution than each of the plurality of images.

2. The apparatus according to claim 1, wherein the image correction unit corrects image data, of each correction target image, corresponding to a part of the fluorescent sample whose fluorescent coloring agent is degraded by fading.

3. The apparatus according to claim 1, wherein the image correction unit comprises:

a histogram generation unit generating a histogram of image data, of each of the plurality of images, for each relative position;

a lookup table generation unit generating a lookup table by comparing the histograms; and

a gray-scale correcting unit correcting a gray scale level of each correction target image using the lookup table.

4. The apparatus according to claim 1, wherein the computer readable instructions, when executed by the processor, further implement:

a subject identifying unit identifying each subject contained in the fluorescent sample from image data of each of the plurality of images;

wherein

the image correction unit corrects a gray scale level of a portion corresponding to the subject in the image data using a histogram for each subject of the image data.

5. The apparatus according to claim 1, wherein the computer readable instructions, when executed by the processor, further implement:

an unmixing unit generating, from image data of each of the plurality of images, image data for each fluorescent coloring agent contained in the fluorescent sample;

wherein

the image correction unit corrects the gray scale level of each correction target image using the histogram of the image data generated for each fluorescent coloring agent.

6. The apparatus according to claim 5, wherein the computer readable instructions, when executed by the processor, further implement:

a fluorescence information storage unit storing wavelength data of fluorescence emitted from a fluorescent coloring agent;

wherein

the unmixing unit acquires wavelength data of the fluorescence emitted from the fluorescent coloring agent contained in the fluorescent sample from the fluorescence information storage unit, and generates image data for each fluorescent coloring agent contained in the fluorescent sample using the wavelength data.

7. The apparatus according to claim 1, further comprising:

a microscope for observing the fluorescent sample;

wherein

the imaging unit captures plurality of images of the fluorescent sample which has passed the microscope.

8. The apparatus according to claim 7, wherein the imaging apparatus is a digital camera for a microscope.

10. The imaging apparatus according to claim 1, wherein the image correction unit corrects gray-scale levels data so as to match a histogram of the reference image, which is captured at a reference position, with a histogram of each correction target image captured at a position other than the reference position.

11. The imaging apparatus according to claim 1, wherein

the reference image is a result of first acquiring an image of the fluorescence sample by the imaging unit, and

each correction target image is acquired after acquiring the reference image.

9. An image generating method, comprising:

changing a relative position of an imaging unit for a plurality of images of a fluorescent sample and capturing each of the plurality of images of the fluorescent sample at a different relative position;

setting, as a reference image, one of the plurality of images acquired at one of the different relative positions and correcting a gray-scale level of of at least one correction target image selected from the plurality of images other than the reference image so that each histogram of each respective correction target image other than the reference image matches a histogram of the reference image; and

combining the reference image and each correction target image whose gray-scale levels have been corrected, so as to generate image data with a higher resolution than each of the plurality of images.

12. The method according to claim 9, wherein correcting a gray-scale level of each of the correction target image comprises:

generating a lookup table by comparing the histogram of the reference image data and the histogram of the respective correction target image data; and

correcting a gray scale of image data of the respective correction target image using the lookup table.

13. The method according to claim 12, wherein:

the plurality of different relative positions includes one reference position and shifted positions other than the reference position;

the lookup table is generated for each shifted position by comparing the histogram of the image at the reference position with the histogram of the image at the shifted position.

14. The method according to claim 13, wherein correcting a gray-scale level of each correction target image refers to correcting the gray scale of the respective correction target image at the shifted position, and matching the histogram of the respective correction target image at the shifted position whose gray scale has been corrected with the histogram of the image the reference position.

15. The method according to claim 9, wherein:

correcting a gray-scale level refers to matching a histogram of the reference image, which is captured at a reference position, with a histogram of each correction target image captured at a position other than the reference position.