Assembly for light sheet microscopy

An arrangement for light sheet microscopy comprising:

• a sample vessel for receiving a sample that is located in a medium, the sample vessel being oriented with respect to a plane reference surface;
• illumination optics with an illumination objective for illuminating the sample with a light sheet;
• detection optics with a detection objective;
• and
• a separating layer system with one or more layers of predetermined thickness and made of predetermined materials for spatially separating the medium in which the sample is found from the illumination objective and the detection objective;
• wherein an optical axis of the illumination objective and the light sheet lies in a plane that forms a nonzero illumination angle β with a normal of the reference surface;
• wherein an optical axis of the detection objective forms a nonzero detection angle δ with the normal of the reference surface;
• wherein, at least in a region which is accessible to the illumination objective and the detection objective for illumination and detection, the separating layer system contacts the medium with an interface that is oriented parallel to the reference surface;
• and
• wherein the illumination angle β and detection angle δ are predetermined based on numerical apertures NAD, NAB of the detection objective and of the illumination objective, respectively;
• wherein the numerical aperture NAB of the illumination objective is less than the numerical aperture NAD of the detection objective, and the illumination angle β is greater than the detection angle δ;
• wherein the illumination optics, the detection optics, or both include corrective means for reducing aberrations arising as a result of the oblique passage of illumination light and/or the light to be detected through interfaces of the separating layer system;
• wherein the separating layer system comprises:
• a plate-shaped or sheet-shaped covering that covers the sample vessel, the plate-shaped or sheet-shaped covering comprising a predetermined material and having a predetermined thickness;
• wherein a first surface of the plate-shaped or sheet-shaped covering is in contact with the medium at least in the region that is accessible to the illumination objective and detection objective for illumination and detection;
• wherein a second surface of the covering is in contact with a gas or an immersion medium, as further component of the separating layer system, at least in the region that is accessible to the illumination objective and detection objective for illumination and detection;
• wherein the predetermined material for the covering has a refractive index that differs by less than 5% from a refractive index of the medium in which the sample is located;
• and
• wherein the illumination objective and the detection are arranged below the sample vessel.