Patent Document ID: 10001634
Application ID: 14412508
Patent Status: 1

Claim One:
1. A method for preparing for and carrying out the microscopic acquisition of image stacks of a sample from various orientation angles (ai), where i=1,. .. , N, and where N is a natural number, the method comprising: prior to acquisition of the image stacks the following preparation steps for determining a section volume: placing the sample in a sample holder, defining a sample coordinate system (X p , Yp, Zp) in the sample holder, wherein the sample holder can be translated in a space spanned by a corresponding detection coordinate system (X D , Y D , Z D ) and can be rotated about at least one axis of rotation parallel to the axis (Yp) of the sample coordinate system, and wherein a detection direction lies parallel to the axis (Z D ) and the position of the two coordinate systems relative to each other is known; acquiring a projection image for at least two orientation angles (al, a2) in the detection coordinate system along the detection direction, the projection image being generated by illuminating the sample, and as the first orientation angle (al) is set; a) selecting a sample region to be examined, wherein the size of an image field is chosen such that the sample region to be examined is located, in a projection along the detection direction, at least partially in the region of the image field; b) at the first orientation angle (al) defining a first region of interest which overlaps at least partially with the image field; c) determining the position of the image field centre in the detection coordinate system in a plane perpendicular to the detection direction; d) determining a set of coordinates in the detection coordinate system depending on the shape of the first region of interest, wherein the set of coordinates is sufficient to clearly define the position and the dimensions of the first region of interest in the image field and thus in the detection coordinate system; e) determining a first sample volume in the sample coordinate system from the position of an image field centre of the projection image, from the set of coordinates, and from coordinates in the detection direction perpendicular to the first region of interest; setting the second orientation angle (a2) and rotating the sample about the at least one axis of rotation by the angular difference between the two orientation angles, (a2−al), and wherein the sample is optionally translated in the detection coordinate system so that the sample region to be examined is located at least partially in the region of the image field; performing steps a) to d) at the second orientation angle (a2), defining a second region of interest, and determining the position of the image field centre, a further set of coordinates in the detection coordinate system and a second sample volume; determining a section volume in which the two sample volumes overlap; and after the section volume has been determined, determining the parameters in the sample coordinate system for image stack acquisition for the orientation angles (ai) with reference to the section volume, including initial and final positions for the stack acquisition along the detection direction for each orientation angle (ai) and the position of the section volume relative to the axis of rotation; positioning the sample correspondingly in the detection coordinate system, and carrying out the image stack acquisition for each orientation angle (ai).