Variable Image Splitter for High-Resolution Microscopy
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Solution Overview
Problem
Existing methods for high-resolution microscopy, particularly in the z-direction, face challenges such as distortions, changes in imaging scale, and limitations in adjusting beam splitting, leading to inaccurate localization of molecules and requiring refocusing when measuring objects near the cover glass.
Innovation Solution
A variable image splitter module that splits the microscope image into multiple partial images on a sensor, allowing for precise adjustment of object planes without shifting the focal point into the cover glass, using movable prisms and beam-splitter cubes to maintain focus and alignment across different microscope objectives.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If beam splitting is used to achieve multiple object planes, then measurement capability in z-direction is improved, but imaging scale changes and focus shifts occur
Solution Approach 1:
A movable prism is introduced as an intermediary element in the optical path to adjust the beam splitting ratio. The prism's position can be varied to control the degree of beam splitting, thereby adjusting the separation between object planes while maintaining stable imaging scale and focus through compensation mechanisms.
Solution Approach 2:
The beam splitting ratio is made variable by changing the position of the movable prism. This parameter change allows dynamic adjustment of the object plane separation to optimize z-direction measurement while compensating for associated focus shifts to maintain imaging scale stability.
2Adaptability or versatility
If beam splitting is adjusted for different object planes, then z-direction measurement range is improved, but focus alignment deteriorates
Solution Approach 1:
A feedback mechanism is implemented to monitor focus alignment when the movable prism is adjusted. The system detects focus shifts caused by beam splitting changes and automatically compensates by adjusting other optical parameters to maintain accurate focus alignment across different object planes.
Solution Approach 2:
The optical system is made dynamic by introducing a movable prism that can be repositioned to adjust beam splitting for different object planes. This dynamic adjustment capability allows the system to adapt to various measurement requirements while maintaining focus alignment through coordinated movement of other optical elements.
3Measurement precision
If multiple individual images are taken for molecule localization, then localization precision is improved, but image acquisition time increases
Solution Approach 1:
The system transitions from two-dimensional image capture to three-dimensional measurement by utilizing beam splitting to create multiple object planes. This allows simultaneous acquisition of information from different z-depths, effectively reducing the total number of images needed while maintaining or improving localization precision through the added dimensional information.
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 accurate three-dimensional localization of molecules with enhanced spatial resolution, reducing calculation time and improving image quality by maintaining focus and alignment across various object planes without the need for secondary focusing.
Implementation Method 1
A variable image splitter module that splits the microscope image into multiple partial images on a sensor, allowing for precise adjustment of object planes
Implementation Method 2
using movable prisms and beam-splitter cubes to maintain focus and alignment across different microscope objectives
Implementation Method 3
Excitation of the activated molecules to emit fluorescent radiation
Data Source
AI summary
The invention relates to a microscope having an illumination beam path with wide field illumination of a sample and a first detection beam path having a spatially resolved surface receiver, which is reached by a first part of the detection light coming from the sample via the first detection beam path, or an image divider assembly for a microscope. In order to lengthen the optical path length, at least a second part of the detection light coming from the sample is masked out of the detection beam path and, via deflection means belonging to the detection beam path, is led into a second detection beam path and, preferably via further deflection means, is deflected back in the direction of the detection in such a way that detection light is applied to at least two partial regions beside one another on the surface receiver. At least the second part of the detection light runs in an optical element having an optical density that is increased as compared with the first detection beam path, in order to lengthen the optical path length, and the optical element is designed to be displaceable at an angle, preferably perpendicular, to the optical axis of the first detection beam path in order to adjust the optical path length, and has flat surfaces, at least on the light entry and light exit side thereof; a prism is provided, preferably a glass prism, preferably at least in the second detection beam path after a first beam deflection, for deflection in a direction parallel to the first detection beam path, in order to increase the path length and for reverse deflection.