TECHNICAL FIELD

The present disclosure relates to a sample observation device and a sample observation method.

BACKGROUND ART

As a method for observing the inside of a sample having a three-dimensional stereoscopic structure such as a cell, selective plane illumination microscopy (SPIM) is known. For example, a tomographic image observation device described in Patent Document 1 discloses the basic principle of SPIM of emitting planar light to a sample, forming an image of fluorescent light or scattered light generated inside the sample on an imaging surface, and acquiring image data observed from inside the sample.

As another sample observation device using planar light, for example, there is an SPIM microscope described in Patent Document 2. In this conventional SPIM endoscope, planar light is emitted while maintaining a constant inclination angle with respect to an arrangement face of a sample, and observation light from the sample is captured by an observation optical system having an observation axis orthogonal to an emission face of the planar light.

CITATION LIST

Patent Literature

Patent Literature 1: Japanese Unexamined Patent Publication No. S62-180241

Patent Literature 2: Japanese Unexamined Patent Publication No. 2014-202967

SUMMARY OF INVENTION

Technical Problem

In the sample observation device described in Patent Document 2 described above, by emitting planar light onto the whole surface of a focused face of an observation optical system, an image of a fault plane in an observation axial direction can be acquired by performing imaging once. Accordingly, in order to acquire three-dimensional information of a sample, it is necessary to scan the sample in the observation axial direction and acquire images of a plurality of fault planes in the observation axial direction. In such a conventional sample observation device, it is necessary to repeat selection of a fault plane from which an image is acquired (scanning a sample and stopping) and image acquisition until the images of all the fault planes are acquired. In addition, in a case in which an area occupied by an observation target is larger than an imaging area, in addition to the operation of acquiring a cross-sectional image in the observation axial direction, operations of moving a stage in a direction different from the observation axial direction and selecting an imaging field of view and the like are necessary. For this reason, there is a problem in that it takes time until observation image data is acquired.

An object of an embodiment is to provide a sample observation device and a sample observation method.

Solution to Problem

According to one aspect of an embodiment, there is provided a sample observation device including: an emission optical system for emitting planar light onto a sample; a scanning unit for scanning the sample with respect to an emission face of the planar light; an imaging optical system having an observation axis inclined with respect to the emission face and for forming an image from observation light generated in the sample in accordance with the emission of the planar light; an image acquiring unit for acquiring a plurality of partial image data corresponding to a part of an optical image according to the observation light formed as an image by the imaging optical system; and an image generating unit for generating observation image data of the sample based on the plurality of partial image data generated by the image acquiring unit.

In this sample observation device, a sample is scanned by the emission face of the planar light, and the observation axis of the imaging optical system is inclined with respect to the emission face of the planar light. For this reason, the image acquiring unit can sequentially acquire partial image data of fault planes in the direction of the optical axis of the planar light, and the image generating unit can generate observation image data of the sample based on the plurality of partial image data. In this sample observation device, an operation of selecting a field of view is not necessary, and a scanning operation for a sample and image acquisition can be simultaneously performed, whereby the throughput until the acquisition of the observation image data is improved.

In addition, the sample may be held by a sample container having an input face of the planar light, and an optical axis of the planar light according to the emission optical system may be disposed to be orthogonal to the input face of the sample container. In such a case, a plurality of samples can be scanned together using the sample container. In addition, by configuring the optical axis of the planar light to be orthogonal to the input face of the sample container, position correction for the partial image data acquired by the image acquiring unit and the like are not necessary, and the process of generating observation image data can be easily performed.

In addition, the scanning unit may scan the sample in a direction orthogonal to the optical axis of the planar light according to the emission optical system. In such a case, image processing such as position correction for the partial image data acquired by the image acquiring unit and the like is not necessary, and the process of generating observation image data can be easily performed.

In addition, an inclination angle of the observation axis of the imaging optical system with respect to the emission face of the planar light may be in the range of 10° to 80°. In this range, the resolution of an observed image can be sufficiently secured.

Furthermore, an inclination angle of the observation axis of the imaging optical system with respect to the emission face of the planar light may be in the range of 20° to 70°. In this range, the resolution of an observed image can be more sufficiently secured. In addition, a change in the field of view with respect to the amount of change in the angle of the observation axis can be suppressed, and the stability of the field of view can be secured.

In addition, an inclination angle of the observation axis of the imaging optical system with respect to the emission face of the planar light may be in the range of 30° to 65°. In this range, the resolution of an observed image and the stability of the field of view can be more appropriately secured.

In addition, the image acquiring unit may be configured to include a two-dimensional imaging device and for extracting image data corresponding to a part of the optical image of the observation light from data output from the two-dimensional imaging device as the partial image data. According to such a configuration, the partial image data can be acquired with high accuracy.

In addition, the image acquiring unit may include a line sensor for capturing a part of the optical image according to the observation light and outputting the partial image data. According to such a configuration, the partial image data can be acquired with high accuracy.

In addition, the image acquiring unit may include a slit transmitting a part of an optical image according to the observation light and an optical detector for detecting an optical image transmitted through the slit and is configured to generate the partial image data based on data output from the optical detector. According to such a configuration, the partial image data can be acquired with high accuracy.

In addition, the image generating unit may be configured to generate observation image data of the sample on a face orthogonal to the optical axis of the planar light based on the plurality of partial image data. In such a case, a cross-sectional image of the sample in which the influence of background is suppressed can be acquired as an observed image.

In addition, the sample observation device may further include an analysis unit for analyzing the observation image data and generating an analysis result. Since the observation image data generated by the image generating unit is analyzed by the analysis unit, the throughput of the analysis can be improved as well.

In addition, according to one aspect of an embodiment, there is provided a sample observation method including: an emission step of emitting planar light onto a sample; a scanning step of scanning the sample with respect to an emission face of the planar light; an imaging step of forming an image from observation light generated in the sample in accordance with the emission of the planar light using an imaging optical system having an observation axis inclined with respect to the emission face; an image acquiring step of acquiring a plurality of partial image data corresponding to a part of an optical image according to the observation light formed as an image by the imaging optical system; and an image generating step of generating observation image data of the sample based on the plurality of partial image data.

In this sample observation method, a sample is scanned by the emission face of the planar light, and the imaging optical system of which the observation axis is inclined with respect to the emission face of the planar light is used. For this reason, in the image acquiring step, partial image data of fault planes in the direction of the optical axis of the planar light can be sequentially acquired, and in the image generating step, observation image data of the sample can be generated based on a plurality of partial image data. In this sample observation method, an operation of selecting a field of view is not necessary, and a scanning operation for a sample and image acquisition can be simultaneously performed, whereby the throughput until the acquisition of the observation image data is improved.

Advantageous Effects of Invention

According to a sample observation device and a sample observation method, throughput until acquisition of observation image data is improved.

