Method and device for determining a critical angle of an excitation light beam
First Claim
1. A method for determining a critical angle of total reflection, comprising:
- illuminating a sample with an excitation light beam of a light source;
using an image capturing sensor, capturing fluorescence images at a plurality of different angles of incidence of the excitation light beam, wherein each of the fluorescence images comprises a first image area including an object to be examined and a second image area outside the object; and
using a processor, determining a critical angle of total reflection at an interface of the sample based upon an analysis of the fluorescence images, wherein said analysis comprises;
forming for each fluorescence image an intensity ratio between a first fluorescence intensity in the first image area of one of the fluorescence images and a second fluorescence intensity in the second image area of said one of the fluorescence images, wherein said ratio being formed by dividing one of the first and second fluorescence intensities by the other of the first and second fluorescence intensities; and
evaluating the intensity ratios for the plurality of angles of incidence.
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Accused Products
Abstract
A method for determining a critical angle of total reflection based upon images captured at different angles of incidence of a light beam includes illuminating a sample with an excitation light beam, capturing images of at least part of the sample at a plurality of different angles of incidence of the excitation light beam, and determining a critical angle of total reflection at an interface of the sample based upon analysis of the images. An apparatus for determining a critical angle of total reflection at an interface of a sample includes a light source arrangement to illuminate a sample with an angle of incidence, an image capturing arrangement to capture an image of the sample, and a processing arrangement to determine the critical angle of total reflection at an interface of the sample on the basis of an analysis of images captured at a plurality of different angles of incidence.
40 Citations
22 Claims
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1. A method for determining a critical angle of total reflection, comprising:
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illuminating a sample with an excitation light beam of a light source; using an image capturing sensor, capturing fluorescence images at a plurality of different angles of incidence of the excitation light beam, wherein each of the fluorescence images comprises a first image area including an object to be examined and a second image area outside the object; and using a processor, determining a critical angle of total reflection at an interface of the sample based upon an analysis of the fluorescence images, wherein said analysis comprises; forming for each fluorescence image an intensity ratio between a first fluorescence intensity in the first image area of one of the fluorescence images and a second fluorescence intensity in the second image area of said one of the fluorescence images, wherein said ratio being formed by dividing one of the first and second fluorescence intensities by the other of the first and second fluorescence intensities; and evaluating the intensity ratios for the plurality of angles of incidence. - View Dependent Claims (2, 3, 4, 5, 6, 7, 17)
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8. An apparatus for determining a critical angle of total reflection at an interface of a sample, comprising:
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a light source operable to illuminate a sample with an angle of incidence; an image capturing sensor operable to capture fluorescence images of the sample, wherein each of the fluorescence images comprises a first image area including an object to be examined and a second image area outside the object; and a processor operable to determine a critical angle of total reflection at an interface of the sample based upon an analysis of the fluorescence images captured at a plurality of different angles of incidence, wherein said analysis comprises; forming for each fluorescence image an intensity ratio between a first fluorescence intensity in the first image area of one of the fluorescence images and a second fluorescence intensity in the second image area of said one of the fluorescence images, wherein said ratio being formed by dividing one of the first and second fluorescence intensities by the other of the first and second fluorescence intensities; and evaluating the intensity ratios for the plurality of angles of incidence. - View Dependent Claims (9, 10, 11, 12, 13, 19, 20, 21)
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14. A method for determining a critical angle of total reflection, comprising:
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illuminating a sample with an excitation light beam of a light source; using an image capturing sensor, capturing fluorescence images of at least part of the sample at a plurality of different angles of incidence of the excitation light beam and for at least a first image plane and a second image plane different from the first image plane, wherein each image plane comprises a first image area including an object to be examined and a second image area outside the object; and using a processor, determining a critical angle of total reflection at an interface of the sample based upon an analysis of the fluorescence images, wherein said analysis comprises; forming for each image plane an intensity ratio between a first fluorescence intensity in the first image area of one of the image planes and a second fluorescence intensity in the second image area of said one of the image planes, wherein said ratio being formed by dividing one of the first and second fluorescence intensities by the other of the first and second fluorescence intensities; and evaluating the intensity ratios for the plurality of angles of incidence. - View Dependent Claims (15, 16, 18, 22)
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Specification