Orthogonal Illumination via Single Objective Deflection
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Solution Overview
Problem
Existing SPIM microscopy methods face challenges in achieving quickly adjustable, reliable, and versatile orthogonal illumination, often requiring complex and expensive setups, and struggle with sample preparation and shadowing issues due to the vertical arrangement of lenses.
Innovation Solution
A method and device that generate a light strip by passing a light bundle through a conventional microscope objective and using a deflection device to direct the light strip at an angle other than zero degrees, preferably a right angle, to the optical axis, allowing for area-like illumination perpendicular to the observation direction using existing microscope components.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If a vertical arrangement of two objectives is used to achieve thin light strip illumination, then the numerical aperture can be high and working distance large, but the device complexity increases and shadowing occurs
Solution Approach 1:
Instead of using a vertical arrangement of two objectives (illumination objective and detection objective) as in conventional SPIM, the invention uses a single objective with the light strip generated and directed through its entrance pupil. The illumination path is inverted by using a beam deflection device to direct the light strip orthogonally through the objective, eliminating the need for a separate vertical illumination objective and reducing device complexity while maintaining light strip quality
Solution Approach 2:
The single microscope objective serves dual functions: it acts as both the detection objective for collecting fluorescent light and as the optical element through which the illumination light strip is directed. This multi-functionality eliminates the need for separate illumination and detection objectives, simplifying the overall device architecture while maintaining imaging performance
2Illumination intensity
If a vertical arrangement of two objectives is used, then light strip illumination is achieved, but shadowing in the sample occurs
Solution Approach 1:
The invention inverts the conventional illumination geometry by directing the light strip orthogonally through the objective's entrance pupil rather than from above. This orthogonal entry of illumination light through the objective eliminates shadowing effects that occur with vertical illumination, as the light enters the sample from the detection side without being blocked by the objective or mounting structures
Solution Approach 2:
The illumination geometry is changed from a vertical dimension (top-down illumination) to an orthogonal dimension (side illumination through the objective). By entering the sample from the lateral direction through the objective, the light strip illuminates the sample without casting shadows that would be created by vertical illumination geometry
3Ease of manufacture
If conventional microscope objectives are used, then cost-effectiveness is maintained, but orthogonal illumination is difficult to achieve
Solution Approach 1:
The invention makes conventional microscope objectives versatile by enabling them to perform both detection and orthogonal illumination functions. By using the objective's entrance pupil as the illumination path and employing a beam deflection device, standard objectives can achieve SPIM-style orthogonal illumination without requiring specialized custom-designed objectives, thereby maintaining cost-effectiveness while gaining functional versatility
Solution Approach 2:
The illumination geometry is made dynamically adjustable through the use of a beam deflection device (such as a galvanometer mirror or acousto-optic deflector) that can rapidly change the angle and position of the light strip entering the objective. This dynamic control allows flexible illumination patterns and scanning capabilities using conventional objectives, enhancing adaptability without requiring complex fixed optical arrangements
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
Enables flexible and reliable orthogonal illumination, reducing shadowing and sample preparation complexities, while utilizing conventional microscope objectives to maintain cost-effectiveness and simplicity.
Implementation Method 1
generating a light bundle and producing a light strip from the light bundle with an optical means
Implementation Method 2
the light strip after deflection for illuminating the sample spreads at an angle other than zero degrees, in particular a right angle, to the optical axis of the lens and is preferably focused in the sample
Implementation Method 3
Passing the light strip through at least one objective which has optics which are designed and intended to supply detection light
Implementation Method 4
Deflection of the light strip with a deflection device connected downstream of the optics of the lens
Data Source
Figure 1a~1c
Figure 2a~2c
Figure 3a~3c
AI summary
The illumination method involves generating a light beam (1) and generating a strip of light of the light beam through passing the light beam in interaction with an optical medium, where the strip of light pass through a lens (4) having an optic (4.2), which is designed and intended to directly or indirectly supply outgoing detection light from a sample to a detector. The light strip is deflected with a subsequent deflection of the lens such that the strip of light disperses in an angle different from zero degrees, particularly in a right angle to the optical axis of the lens. Independent claims are included for the following: (1) an arrangement for illuminating a sample with selective plane illumination microscope-microscopy, particularly for performing an illumination method using a microscope; and (2) a microscope, particularly a scanning microscope or a confocal scanning microscope or an experimental setup for imaging a sample.