Method, Device and Laser Scanning Microscope for Generating Rasterized Images
AssigneeAbberior Instruments GmbH
InventorsAndreas Schoenle, Lars Kastrup
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Abstract
In order to generate rasterized images of a sample, a pixel size of image points of a rasterized image is set and photons emitted out of the sample which were detected, and for each of which a position of an effective local excitation of the sample for emitting the respective detected photon has been recorded are assigned to that image point of the rasterized image into which the position of the effective local excitation recorded for the respective detected photon falls. To set the pixel size of the image points to an optimized pixel size, the positions of the effective local excitation of the sample for emitting the detected photons are evaluated.
Claims19
A method of generating rasterized images of a sample, comprising the steps:
- setting a pixel size of image points of a rasterized image;
- locally exciting the sample for emitting photons at a position of an effective local excitation;
- scanning the sample with the position of the effective local excitation;
- detecting the photons emitted out of the sample;
- recording the position of the effective local excitation of the sample for each detected photon;
- and
- assigning the photons to that particular image point of the rasterized image into which the position of the effective local excitation recorded for the particular detected photon falls;
- wherein the positions of the effective local excitation of the sample for emitting the detected photons are evaluated to set the pixel size of the image points to an optimized pixel size.
An apparatus for generating rasterized images of a sample comprising
- a data input for data relating to positions of an effective local excitation of the sample for emitting particular detected photons,
- a data output configured to output image data of rasterized images, and
- a data processing unit arranged between the data input and the data output, and configured to execute the following steps:
- setting a pixel size of image points of a rasterized image,
- assigning photons,
- which were emitted from the sample,
- which were detected, and
- for which a position of an effective local excitation of the sample for emitting the particular detected photon was recorded,
- to that particular image point of the rasterized image into which the position of the effective local excitation recorded for the particular detected photon falls, and
- evaluating the positions of the effective local excitation of the sample for emitting the detected photons to set the pixel size of the image points to an optimized pixel size.
A laser scanning microscope for generating rasterized images of a sample comprising
- an emission excitation light source configured to provide an emission excitation light beam of emission excitation light,
- an emission inhibition light source configured to provide an emission inhibition light beam of emission inhibition light,
- an objective configured to focus the emission excitation light beam from the emission excitation light source and the emission inhibition light beam from the emission inhibition light source into a focus region in the sample, wherein an intensity minimum of the emission inhibition light surrounded by intensity maxima of the emission inhibition light and spatially coinciding with an intensity maximum of emission excitation light is formed in the sample,
- a scanning device configured to scan the sample with the emission inhibition light intensity minimum,
- a detector configured to detect photons emitted out of the sample,
- a device configured to, for every photon detected by the detector, record a position of the effective local excitation of the sample for emitting the respective detected photon in the region of the emission inhibition light intensity minimum, and
- an apparatus for generating rasterized images of a sample comprising
- a data input for data relating to the positions of the effective local excitation of the sample for emitting a particular detected photon,
- a data output configured to output image data of the rasterized images, and
- a data processing unit arranged between the data input and the data output, and configured to execute the following steps:
- setting a pixel size of image points of a rasterized image,
- assigning photons,
- which were emitted from the sample,
- which were detected, and
- for which a position of an effective local excitation of the sample for emitting the particular detected photon was recorded,
- to that particular image point of the rasterized image into which the position of the effective local excitation recorded for the particular detected photon falls, and
- evaluating the positions of the effective local excitation of the sample for emitting the detected photons to set the pixel size of the image points to an optimized pixel size.
Dependencies are inferred from the claim text.