Resumen
The invention describes a microscope and method for obtaining 3d images of several transparent or semi-transparent samples basically comprising: causing, while maintaining a constant acquisition angle, a relative displacement according to the direction of detection between the illumination sheet and the sample; obtaining, for said acquisition angle, a single 2d projection image formed by a representative parameter for each pixel; modify the acquisition angle by means of a relative rotation between the illumination sheet and the sample combined with a relative vertical translation between the illumination sheet and the sample, and repeat the previous steps; and generate, from the set of 2d projection images obtained, a 3d image of each of the samples. (Machine-translation by Google Translate, not legally binding)
Reivindicaciones
1. Method for generating 3D images of a collection of samples by means of a microscope, characterized in that the microscope comprises an image acquisition means facing the collection of samples according to a detection direction, and a lighting means configured for emitting a plane sheet of light towards the collection of samples according to a direction perpendicular to the detection direction, where the collection of samples comprises various samples located one on top of another, the method comprising the following steps: - causing a relative movement according to the detection direction between the sheet of light and the sample while maintaining a constant angle of acquisition; - obtaining, for said angle of acquisition, a single 2D projection image formed by a representative parameter for each pixel; - modifying the angle of acquisition by means of a relative rotation between the sheet of light and the sample, combined with a relative vertical translation between the sheet of light and the sample, and repeating the previous steps; and - generating a 3D image of each of the samples from the set of 2D projection images that are produced. 2. Method according to claim 1, wherein the representative parameter for each pixel is a statistical parameter. 3. Method according to claim 2, wherein the statistical parameter is selected from the maximum value, variance, minimum value, standard deviation, average value and correlation between pixels. 4. Method according to any of claims 2-3, wherein the statistical parameter is produced by causing a continuous relative movement between the sheet of light and the sample according to the detection direction and acquiring images at a high speed. 5. Method according to claim 1, wherein the representative parameter for each pixel is the sum of the intensities of each pixel. 6. Method according to claim 5, wherein the sum of the intensities of each pixel is produced by causing a continuous relative movement between the sheet of light and the sample according to the detection direction while at the same time the exposure of the image acquisition means is kept open. 7. Method according to any of the preceding claims, wherein the step of modifying the angle of acquisition further comprises modifying the position of the sample within a plane perpendicular to the axis of rotation of said sample in order to always center the same portion of the sample opposite the detection direction. 8. Method according to any of the preceding claims, wherein the step of generating a 3D image of the sample comprises applying a filtered backprojection algorithm to the set of projection images that are produced. 9. Method according to any of claims 1-7, wherein the step of generating a 3D image of the sample comprises applying the inverse Radon transform to the set of projection images that are produced. 10. Method according to any of the preceding claims, which further comprises combining several 3D images that are produced by using different representative parameters for producing an enhanced 3D image. 11. Microscope for generating 3D images of a collection of samples capable of carrying out the method of any of the preceding claims, characterized in that it comprises an image acquisition means facing the collection of samples according to a detection direction, and a lighting means configured for emitting a plane sheet of light towards the collection of samples according to a direction perpendicular to the detection direction, wherein the collection of samples comprises various samples located one on top of another, further comprising: - means for causing a relative movement according to the detection direction between the sheet of light and the sample while maintaining a constant angle of acquisition; - means for obtaining, for said angle of acquisition, a single 2D projection image formed by a representative parameter for each pixel; - means for modifying the angle of acquisition by means of a relative rotation between the sheet of light and the sample, combined with a relative vertical translation between the sheet of light and the sample; and - means for generating a 3D image of each of the samples from the set of 2D projection images that are produced. 12. Microscope according to claim 11, wherein the means for causing a relative movement according to the detection direction between the sheet of light and the sample are configured for performing a continuous movement. 13. Microscope according to any of claims 11-12, wherein the means for performing a relative vertical translation between the sheet of light and the sample comprise an electric motor.
