CA2365093A1 - Inverted darkfield contrast microscope and method - Google Patents
Inverted darkfield contrast microscope and method Download PDFInfo
- Publication number
- CA2365093A1 CA2365093A1 CA002365093A CA2365093A CA2365093A1 CA 2365093 A1 CA2365093 A1 CA 2365093A1 CA 002365093 A CA002365093 A CA 002365093A CA 2365093 A CA2365093 A CA 2365093A CA 2365093 A1 CA2365093 A1 CA 2365093A1
- Authority
- CA
- Canada
- Prior art keywords
- image
- darkfield
- microscope according
- microscope
- inverted
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Classifications
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
- G02B21/365—Control or image processing arrangements for digital or video microscopes
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
- G02B21/0088—Inverse microscopes
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/02—Objectives
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/06—Means for illuminating specimens
- G02B21/08—Condensers
- G02B21/10—Condensers affording dark-field illumination
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
- G02B21/362—Mechanical details, e.g. mountings for the camera or image sensor, housings
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- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Optics & Photonics (AREA)
- Engineering & Computer Science (AREA)
- Multimedia (AREA)
- Computer Vision & Pattern Recognition (AREA)
- Microscoopes, Condenser (AREA)
Abstract
A novel microscope and method of obtaining images includes a combination of the conventional darkfield illuminations technique with electronic image inversion (converting a positive to a negative image) and other improvements to further enhance the contrast and resolution of the final image. The microscope and method are referred to herein as Inverted Darkfield Contrast (IDC) and are believed to be particularly suitable for viewing live cells in real time with no staining or preparation.
Claims (40)
1. A method for microscopy of samples including:
using a microscope with a high numerical aperture darkfield illuminating system with an imaging means to acquire a darkfield image of a sample, electronically inverting the darkfield image to obtain a negative image thereof, and presenting the electronically inverted image as the final image so that the final image is displayed as an apparent brightfield image.
using a microscope with a high numerical aperture darkfield illuminating system with an imaging means to acquire a darkfield image of a sample, electronically inverting the darkfield image to obtain a negative image thereof, and presenting the electronically inverted image as the final image so that the final image is displayed as an apparent brightfield image.
2. A method according to claim 1 wherein the darkfield image is a colour darkfield image.
3. A method according to claim 1 wherein the darkfield image is a black and white image.
4. A method according to claim 1 wherein the darkfield images contains information outside the visible spectral range.
5. A method according to claim 1 wherein the darkfield image contains ultraviolet information.
6. A method according to claim 1 wherein the darkfield image contains infrared information.
7. A method according to claim 1 wherein the darkfield image is collected at a frame rate of at least 25 frames per second.
8. A method according to claim 1 wherein the darkfield image is collected as an analog image.
9. A method according to claim 1 wherein the darkfield image is collected as a digital image.
10. A method according to claim 1 wherein the electronic inversion is carried out using analog circuitry.
11. A method according to claim 1 wherein the electronic inversion is carried out using digital circuitry.
12. A method according to claim 1 wherein the electronic inversion is an integral function of signal processing on a camera head or a camera control unit.
13. A method according to claim 1 wherein a choice as to whether the image is inverted is under the control of 3 supervising computer and can be selective from frame to frame.
14. A method according to claim 1 wherein the system is optimized for use with living biological samples by choosing a darkfield illuminating system with an internal aperture of no less than 1.0 NA and an external aperture exceeding 1.33 by the greatest possible amount.
15. A method according to claim 1 wherein light illuminating the object is limited to include only visible wavelengths of light.
16. A method according to claim 1 wherein light illuminating the object is limited to bandwidths of light which correspond to the best wavelength correction points of an objective lens being used to illuminate the sample.
17. A method according to claim 1 wherein image information is separated into luminance and chrominance components, and the luminance component only is inverted.
18. A method according to claim 1 wherein image information is separated into luminance and chrominance components, and the chrominance component only is inverted.
19. A method according to claim 1 wherein image information is separated into luminance and chrominance components, and both the luminance and chrominance components are inverted.
20. A method according to claim 1 wherein image information is separated into its colour components, and at least one colour component is selectively inverted.
21. A microscope comprising a darkfield illuminating system which illuminates a sample to create a darkf eld image thereof, and electronic inverting means operable to electronically invert the darkfield image to create a resultant brightfield image.
22. A microscope according to claim 21 wherein image sensing means for the darkfield image comprises a colour image sensor.
23. A microscope according to claim 21 wherein image sensing means for the darkfield image comprises a monochromatic or black and white image sensor.
24. A microscope according to claim 21 wherein image sensing means for the darkfield image is capable of imaging information outside the visible spectral, range.
25. A microscope according to claim 21 wherein image sensing means for the darkfield image is capable of imaging ultraviolet information.
26. A microscope according to claim 21 wherein image sensing means for the darkfield image is capable of imaging infrared information.
27. A microscope according to claim 21 wherein the darkfield image is collected at a frame rate of at least 25 frames per second.
28. A microscope according to claim 21 wherein image sensing means for the darkfield image has an analog image output.
29. A microscope according to claim 21 wherein image sensing means for the darkfield image has a digital image output.
30. A microscope according to claim 21 wherein the electronic inverting means uses analog circuitry.
31. A microscope according to claim 21 wherein the electronic ,converting means uses digital circuitry.
32. A microscope according to claim 21 wherein the electronic inverting means is an integral function of signal processing on a camera head or a camera control unit.
33. A microscope according to claim 32 wherein a choice as to whether the image is inverted is under the control of a supervising computer and can be selected from frame to frame.
34. A microscope according to claim 21 wherein the darkfield illumination system is optimized for use with living biological samples by choosing a darkfield illuminating system with an internal aperture of no less and 1:0 NA and an external aperture exceeding 1.33 NA.
35. A microscope according to claim 21 wherein ultraviolet light from a light source is limited from impinging on the sample by a suitable filter means.
36. A microscope according to claim 21 wherein an infrared light from a light source is limited from impinging on the sample by a suitable filter means.
37. A microscope according to claim 21 wherein illuminating light is filtered into several wavelength bands which are chosen to match the wavelength bands of best optical performance of an objective lens being used to form the image of the sample.
38. A microscope according to claim 21 wherein imaging means is located in the first image plane of an objective lens.
39. A microscope according to claim 21 wherein an objective lens is an infinity corrected lens, and imaging means is placed in the first image plane of a tube lens of an infinity corrected optical system.
40. A microscope according to claim 21 also including an infrared filter which removes substantially all of the heat generated by a light source prior to an illuminating beam impinging on the sample.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA002365093A CA2365093C (en) | 1998-07-10 | 1999-07-12 | Inverted darkfield contrast microscope and method |
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA2,243,090 | 1998-07-10 | ||
| CA002243090A CA2243090A1 (en) | 1998-07-10 | 1998-07-10 | Inverted darkfield contrast microscope and method |
| CA002365093A CA2365093C (en) | 1998-07-10 | 1999-07-12 | Inverted darkfield contrast microscope and method |
| PCT/CA1999/000626 WO2000003283A2 (en) | 1998-07-10 | 1999-07-12 | Inverted darkfield contrast microscope and method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2365093A1 true CA2365093A1 (en) | 2000-01-20 |
| CA2365093C CA2365093C (en) | 2010-02-02 |
Family
ID=25680368
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002365093A Expired - Fee Related CA2365093C (en) | 1998-07-10 | 1999-07-12 | Inverted darkfield contrast microscope and method |
Country Status (1)
| Country | Link |
|---|---|
| CA (1) | CA2365093C (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6865013B2 (en) | 2000-12-01 | 2005-03-08 | Auburn University | High-resolution optical microscope |
| US6992819B2 (en) | 2000-12-01 | 2006-01-31 | Auburn University | High-resolution optical microscope for quick detection of pathogens |
| CN113050262A (en) * | 2021-02-05 | 2021-06-29 | 安徽医科大学 | Universal oblique illumination microscopic imaging system and correction algorithm thereof |
| CN116297337A (en) * | 2022-09-09 | 2023-06-23 | 山东大学 | Method for judging number of layers of two-dimensional material by using dark field optical imaging technology |
-
1999
- 1999-07-12 CA CA002365093A patent/CA2365093C/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6865013B2 (en) | 2000-12-01 | 2005-03-08 | Auburn University | High-resolution optical microscope |
| US6992819B2 (en) | 2000-12-01 | 2006-01-31 | Auburn University | High-resolution optical microscope for quick detection of pathogens |
| US7106503B2 (en) | 2000-12-01 | 2006-09-12 | Auburn University | High-resolution optical microscope |
| CN113050262A (en) * | 2021-02-05 | 2021-06-29 | 安徽医科大学 | Universal oblique illumination microscopic imaging system and correction algorithm thereof |
| CN113050262B (en) * | 2021-02-05 | 2023-06-09 | 安徽医科大学 | General oblique illumination microscopic imaging system and correction algorithm thereof |
| CN116297337A (en) * | 2022-09-09 | 2023-06-23 | 山东大学 | Method for judging number of layers of two-dimensional material by using dark field optical imaging technology |
| CN116297337B (en) * | 2022-09-09 | 2023-10-31 | 山东大学 | Method for judging number of layers of two-dimensional material by using dark field optical imaging technology |
Also Published As
| Publication number | Publication date |
|---|---|
| CA2365093C (en) | 2010-02-02 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| MKLA | Lapsed |
Effective date: 20160712 |