Method for imaging in a microscope with oblique illumination
A method for imaging in a microscope with oblique illumination includes illuminating an object by an illumination beam path that is obliquely incident on an object plane of the microscope. A microscopic image of the object and a corresponding digital image signal are produced. The digital image signal is processed by digital image processing using a convolution kernel to increase contrast. An increased-contrast digital image is produced from the processed digital image signal.
1. A method for imaging in a microscope with oblique illumination, the method comprising:
illuminating an object by an illumination beam path that is obliquely incident on an object plane of the microscope;
producing a microscopic image of the object and a corresponding digital image signal;
processing the digital image signal by digital image processing using a convolution kernel to increase contrast; and
producing an increased-contrast digital image from the processed digital image signal,
wherein the convolution kernel is used whose orientation is aligned in relation to the direction of the oblique illumination in order to further enhance the increased contrast.
2. The method according to claim 1 , wherein the convolution kernel is a square convolution kernel.
3. The method according to claim 1 , wherein the convolution kernel is used in the form of a convolution matrix as follows:
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4. The method according to claim 3 , wherein the convolution kernel is used whose orientation is aligned in relation to the direction of the oblique illumination in order to further enhance the increased contrast, and wherein a direction of a numerical sequence, considered as a vector, of the convolution kernel corresponds with the direction from which the oblique illumination strikes the object.
5. The method according to claim 1 , wherein the illumination beam path is produced by illuminating a decentered region in or close to an aperture plane of an illumination assembly of the microscope.
6. The method according to claim 5 , wherein the illuminated region is selected to be larger than is necessary for imaging with oblique illumination.
7. The method according to claim 6 , wherein the illuminated region is selected to be 100% larger than is necessary for imaging with oblique illumination.
8. The method according to claim 5 , wherein the illuminated region is selected to be up to one-half the pupil size of an entrance pupil, arranged to be conjugate thereto, of the microscope objective.
9. The method according to claim 5 , wherein the illuminated decentered region and/or the convolution kernel used is determined as a function of the microscope objective used.
10. The method according to claim 2 , wherein the convolution kernel is symmetrical.
11. A method for imaging in a microscope with oblique illumination, the method comprising:
illuminating an object by an illumination beam path that is obliquely incident on an object plane of the microscope;
producing a microscopic image of the object and a corresponding digital image signal;
processing the digital image signal by digital image processing using a convolution kernel to increase contrast; and
producing an increased-contrast digital image from the processed digital image signal,
wherein the convolution kernel is a square convolution kernel.
12. The method according to claim 11 , wherein the convolution kernel is symmetrical.
13. The method according to claim 11 , wherein the illumination beam path is produced by illuminating a decentered region in or close to an aperture plane of an illumination assembly of the microscope, and wherein the illuminated decentered region is selected to be 100% larger than is necessary for imaging with oblique illumination.
14. The method according to claim 11 , wherein the illumination beam path is produced by illuminating a decentered region in or close to an aperture plane of an illumination assembly of the microscope, and wherein the illuminated region is selected to be up to one-half the pupil size of an entrance pupil, arranged to be conjugate thereto, of the microscope objective.
15. The method according to claim 11 , wherein the illumination beam path is produced by illuminating a decentered region in or close to an aperture plane of an illumination assembly of the microscope, and wherein the illuminated decentered region and/or the convolution kernel used is determined as a function of the microscope objective used.
16. A method for imaging in a microscope with oblique illumination, the method comprising:
illuminating an object by an illumination beam path that is obliquely incident on an object plane of the microscope;
producing a microscopic image of the object and a corresponding digital image signal;
processing the digital image signal by digital image processing using a convolution kernel to increase contrast; and
producing an increased-contrast digital image from the processed digital image signal,
wherein the convolution kernel is used in the form of a convolution matrix as follows:
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17. The method according to claim 16 , wherein the convolution kernel is used whose orientation is aligned in relation to the direction of the oblique illumination in order to further enhance the increased contrast, and wherein a direction of a numerical sequence, considered as a vector, of the convolution kernel corresponds with the direction from which the oblique illumination strikes the object.
18. The method according to claim 16 , wherein the illumination beam path is produced by illuminating a decentered region in or close to an aperture plane of an illumination assembly of the microscope, and wherein the illuminated decentered region is selected to be 100% larger than is necessary for imaging with oblique illumination.
19. The method according to claim 16 , wherein the illumination beam path is produced by illuminating a decentered region in or close to an aperture plane of an illumination assembly of the microscope, and wherein the illuminated region is selected to be up to one-half the pupil size of an entrance pupil, arranged to be conjugate thereto, of the microscope objective.
20. The method according to claim 16 , wherein the illumination beam path is produced by illuminating a decentered region in or close to an aperture plane of an illumination assembly of the microscope, and wherein the illuminated decentered region and/or the convolution kernel used is determined as a function of the microscope objective used.