US-12620062-B2 - Method for generating an overview image of an object

US12620062B2US 12620062 B2US12620062 B2US 12620062B2US-12620062-B2

Abstract

Generating an overview image of an object includes, during a first capture duration, scanning regions of the object along a trajectory with illumination radiation, where the trajectory is chosen individually on the basis of properties of the object. First images of the object which together with position data (x, y, z) of an absolute distance measurement system are stored as an individual overview image are captured during the scanning of the trajectory. During a second capture duration, at least regions of the object are scanned with illumination radiation, and second images are captured; regions of the object which have already been imaged during the first capture duration are not imaged again. The first images captured during the first capture duration and the second images captured during the second capture duration are merged to form a resultant overview image of the object.

Inventors

Thomas KALKBRENNER
Joerg Engel
Joerg SIEBENMORGEN

Assignees

CARL ZEISS MICROSCOPY GMBH

Dates

Publication Date: 20260505
Application Date: 20240125
Priority Date: 20230131

Claims (12)

1 . A method for generating an overview image of an object, the method comprising: scanning with illumination radiation, during a first capture duration, first regions of the object along a trajectory, wherein the trajectory is chosen individually based on properties of the object; capturing, while scanning the illumination radiation along the trajectory during the first capture duration, first images of the first regions of the object; storing the first images of the object together with position data (x, y, z) of an absolute distance measurement system for the first images as an individual overview image; scanning with illumination radiation, during a second capture duration, at least second regions of the object; capturing, while scanning the illumination radiation during the second capture duration, second images of the second regions of the object, wherein the first regions of the object, images of which that have already been imaged during the first capture duration, are not scanned with illumination and imaged again or are scanned with illumination and imaged again only at a proportion of no more than 50%, during the second capture duration, and wherein the captured second images of the second regions of the object do not overlap the captured first images of the first regions of the object or overlap the captured first images of the first regions of the object at a proportion of no more than 50%; and merging the first images captured during the first capture duration and the second images captured during the second capture duration to form a resultant overview image of the object, wherein merging the first images captured during the first capture duration and the second images captured during the second capture duration to form a resultant overview image of the object is based on the position data (x, y, z) of the absolute distance measurement system for the first images and on position data (x, y, z) of the absolute distance measurement system for the second images.
2 . The method according to claim 1 , wherein the choice of the trajectory and guidance of the illumination radiation along the trajectory during the first capture duration is achieved manually.
3 . The method according to claim 1 , wherein the choice of the trajectory and guidance of the illumination radiation along the trajectory during the first capture duration is achieved by applying an algorithm, wherein a starting point and/or an end point of the trajectory (T) is chosen individually.
4 . The method according to claim 1 , further comprising selecting the second regions to be scanned during the second capture duration based on the first images.
5 . The method according to claim 1 , further comprising: during the first capture duration, assigning at least one additional piece of information to at least one of the first images; and storing the at least one piece of information together with the to at least one of the first images.
6 . The method according to claim 1 , further comprising: determining that the illumination radiation has been guided by a minimum distance along the trajectory since a previous first image was captured; and based on the determination, capturing a further first image.
7 . The method according to claim 1 , wherein the illumination radiation is shaped as a line.
8 . The method according to claim 1 , wherein the illumination radiation is shaped as a light sheet extending in a single plane.
9 . The method according to claim 8 , further comprising: determining that the illumination radiation has been guided by a minimum distance along the trajectory since a previous first image was captured and at an angle greater than zero to the light sheet plane; and based on the determination, capturing a further first image.
10 . The method according to claim 8 , wherein the first images captured during the first capture duration are subjected to a maximum intensity projection of the illumination radiation.
11 . The method according to claim 1 , wherein the first regions of the object, images of which that have already been imaged during the first capture duration, are not scanned with illumination and imaged again or are scanned with illumination and imaged again only at a proportion of no more than 10%, during the second capture duration, and wherein the captured second images of the second regions of the object do not overlap the captured first images of the first regions of the object or overlap the captured first images of the first regions of the object at a proportion of no more than 10%.
12 . The method according to claim 1 , wherein the first regions of the object, images of which that have already been imaged during the first capture duration, are not scanned with illumination and imaged again during the second capture duration, and wherein the captured second images of the second regions of the object do not overlap the captured first images of the first regions of the object.

Description

CROSS REFERENCE TO RELATED APPLICATION This application claims priority under 35 U.S.C. § 119 to German Application No. 10 2023 200 784.3, filed Jan. 31, 2023, the disclosure of which is incorporated herein by reference in its entirety. FIELD OF THE INVENTION This disclosure relates to generating an overview image of an object, in particular, in the field of microscopy. BACKGROUND In the field of microscopy, it is common to fully or partly represent an object to be imaged (also referred to below as a sample) in an overview image before the actual image capture. This is used, for example, to assess the suitability of the object for the application of the imaging method provided and/or in order to identify regions of interests (ROI) of the object in advance. Typically, overview images that have a lower resolution than the images captured by means of the actual imaging method are captured for this purpose. In addition, the lower-resolution images are not suitable, or suitable only to a very limited extent, for being magnified, that is to say for “zooming” in to specific image details. Due to firmly predetermined scanning paths, the capture of an overview image, and also that of an actually desired image, commonly includes significant regions of the object field in which no relevant information is present, but whose image data are nevertheless stored and evaluated. For example, when using predetermined scanning paths, regions of a sample carrier (coverslip, petri dish etc.) on which no sample is located are frequently captured. SUMMARY This disclosure is based on the object of proposing one possibility for generating an overview image of an object, in which magnifications of image details while gaining information are possible and a more efficient capture of an object field can be achieved. The object is achieved by a by the techniques disclosed herein. The disclosed techniques can be used for generating an overview image of an object, in particular, in the field of microscopy. As will be discussed further below, particular advantages can be achieved when the disclosed techniques are applied to the field of light-sheet microscopy (or single-plane illumination microscopy; SPIM). Image data of the object are recorded and processed during a first and a second capture duration. A set of image data and images obtained during the first capture duration and, if appropriate, an image generated therefrom will also be referred to below as an individual overview image. During the first capture duration, regions of the object are scanned along a trajectory using illumination radiation, in particular, shaped illumination radiation, in particular, laser radiation. In the process, the trajectory is chosen individually on the basis of properties, for example, the size, the shape and/or the nature of the object and/or the obtained image information related to the object. In other words, the manifestation of the trajectory is determined individually by each object to be imaged. Repeated use of the method for one at the same object then results in different trajectories. During the scanning of the trajectory, first images of the object are captured, which are stored together with the position data from a global positioning system or an absolute distance measurement system. An absolute distance measurement system enables at any time the unambiguous indication of a position of the point of the object. For example, such an absolute distance measurement system can be aligned with a microscope having a displaceable sample stage. When the object is moved by means of the sample stage and/or shaped illumination radiation used is deflected, the positions thereof in relation to one another can be unambiguously determined at any time. During the performance of the method, at least regions of the object are scanned with, in particular, shaped, illumination radiation during a second capture duration and second images are captured. The regions of the object which have already been imaged during the first capture duration by means of the first images are not imaged again, or are imaged again only at a small proportion. Due to the illumination expanding for reasons of technology and the object region inevitably captured, regions imaged by means of the first images may overlap with regions imaged by means of the second images. It can be desirable that the proportion of such overlaps should be kept as small as possible. The proportion of regions captured both by means of the first images and also by means of the second images should in this case be no more than 50%, advantageously no more than 25%, preferably no more than 10%. In order to obtain a resultant overview image, the first images captured during the first capture duration and the second images captured during the second capture duration are merged to form the resultant overview image (also: systematic overview image) of the object. A systematic overview image, or an overview image captur