Light-sheet Microscopy Device with Inclined Immersion Chamber
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Solution Overview
Problem
Existing light sheet microscopy systems, such as SPIM, are cumbersome and complicate sample movement due to non-parallel camera planes and limited numerical aperture, making them difficult to use with immersion chambers and large samples.
Innovation Solution
A device integrated with an inverted microscope that includes an immersion chamber with a shaping mechanism to transform the light beam into a sheet, allowing it to enter perpendicular to the optical axis, thus maintaining alignment and enabling easier sample movement without limiting numerical aperture.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If an SPIM device is attached to the frame of the inverted microscope with inclined optical paths, then light sheet microscopy can be performed on an inverted microscope, but the XY movements of the sample become complex and the camera plane is not parallel to the XY axes
Solution Approach 1:
Instead of inclining the optical paths as in conventional SPIM, this invention inverts the approach by making the optical path parallel to the vertical axis and inclining the immersion chamber bottom. The light sheet is generated parallel to the optical axis, and the chamber bottom is inclined at an angle (e.g., 45 degrees) relative to the light sheet propagation direction, achieving SPIM functionality with simplified sample movement on the XY stage.
Solution Approach 2:
The invention introduces a new geometric configuration by inclining the immersion chamber bottom in a different dimensional orientation. Rather than changing the light path angle in the horizontal plane, the chamber bottom is inclined in the vertical plane, creating a new spatial relationship that decouples the light sheet generation from sample stage movement complexity.
2Adaptability or versatility
If a reflector is placed near the sample in the focal plane, then light sheet microscopy can be implemented on conventional microscopes, but the proximity limits movement capabilities and the size limits the maximum numerical aperture
Solution Approach 1:
The invention extracts the reflector from its conventional position near the sample in the focal plane and relocates it to a position where the optical path is parallel to the vertical axis. This extraction eliminates the spatial constraints that limited sample movement and numerical aperture, as the light sheet is now generated above the chamber rather than within the focal plane near the sample.
3Ease of operation
If the imaging objective moves, then different sample positions can be observed, but the alignment of the light sheet with the focal plane changes and realignment is required
Solution Approach 1:
Instead of moving the light sheet to follow the objective (conventional approach), this invention inverts the relationship by keeping the light sheet fixed and parallel to the vertical axis while the objective moves vertically along the same axis. The immersion chamber bottom inclination ensures that the light sheet remains aligned with the focal plane regardless of objective position, eliminating the need for realignment.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The solution reduces bulk, simplifies sample movement, increases movement capabilities, and maintains high numerical aperture, allowing for efficient SPIM imaging without realigning the light sheet with the observation plane.
Implementation Method 1
shaping means, arranged to shape the light beam in such a way as: that before transformation by the means of transformation, the beam of light does not have a preferred direction of elongation in a plane perpendicular to its direction of propagation, and that after transformation by the means of transformation, the beam of light has a preferred direction of elongation in a plane perpendicular to its direction of propagation
Data Source
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AI summary
The present invention relates to a device for carrying out light-sheet microscopy comprising: injecting means (203.2); an immersion chamber (205); guiding means (301 to 305) arranged to guide a light beam from the injecting means (203.2) and as far as into the immersion chamber (205) along a guiding optical path; shaping means (302, 303, 305) arranged to shape the light beam so as to give it a shape that is elongate along one spatial dimension so as to create a light sheet in a sheet plane located in the immersion chamber (205); and an objective (202). The shaping means (302, 303, 305) are located along the guiding optical path between the injecting means (203.2) and the immersion chamber (205), and are located between a first plane perpendicular to the optical axis (113) of the objective (202) and passing through the aperture (314) of the immersion chamber (205) and a second plane perpendicular to the optical axis (113) of the objective (202) and located beyond the distal end (312) of the objective (202) relative to the first plane (316).