Adjustable Phase Diaphragm for Meniscus Compensation in Microtiter Plates
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Solution Overview
Problem
Phase-contrast microscopy in microtiter plates is hindered by the curvature of object liquid surfaces, leading to artefacts and reduced contrast due to the meniscus effect, which existing solutions like hydrophobic coatings, specific lid designs, and adjustable phase diaphragms fail to adequately address, especially in microtiter plates with varying well diameters and object liquids.
Innovation Solution
A phase-contrast microscopy method and microscope that uses a changeable optical deflection element to adjust the illumination beam's path and geometry, compensating for the curvature of the object liquid surface by deflecting the beam before it enters the specimen vessel, and employing a Bertrand lens unit to optimize the overlap of direct and specimen radiation, allowing for automatic control of the deflection element's position and shape.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If a standardized microtiter plate with small well diameter is used, then productivity is improved, but the meniscus effect increases causing reduced contrast and image quality
Solution Approach 1:
The patent employs a dynamically adjustable phase diaphragm that can change its opening geometry and position in real-time. This dynamic adjustment compensates for the meniscus effect in different well sizes, maintaining optimal contrast for both small wells in microtiter plates and larger wells, thus resolving the contradiction between productivity and image quality
Solution Approach 2:
The system changes multiple parameters simultaneously: the phase diaphragm opening size, its position relative to the phase ring, and the illumination aperture. These parameter adjustments are coordinated to compensate for meniscus effects across different well diameters, enabling high-quality imaging in both small microtiter plate wells and larger specimen vessels
2Device complexity
If a fixed phase diaphragm opening is used, then device complexity is reduced, but adaptability to different well diameters and object liquids is worsened
Solution Approach 1:
The phase diaphragm is designed with dynamic adjustment capabilities, allowing its opening geometry and position to be modified electronically. This enables the same device to adapt to different well diameters and object liquid properties without requiring multiple fixed configurations, thereby achieving versatility while maintaining reasonable device complexity through automated control
Solution Approach 2:
The adjustable phase diaphragm system serves multiple functions: it optimizes phase contrast for different well sizes, compensates for varying meniscus effects, and adapts to different object liquids. This multi-functionality allows a single device configuration to handle diverse specimen vessels, eliminating the need for multiple specialized setups
3Ease of operation
If the illumination beam is not adjusted for meniscus compensation, then ease of operation is improved, but measurement precision of specimen observation is worsened
Solution Approach 1:
The system performs automatic meniscus compensation through electronic control of the phase diaphragm and illumination parameters. The compensation process occurs automatically without requiring manual intervention from the operator, thus maintaining ease of operation while significantly improving measurement precision and image quality
Solution Approach 2:
The system uses feedback mechanisms to detect and compensate for meniscus effects. By monitoring the actual imaging conditions and adjusting the phase diaphragm parameters accordingly, the system automatically optimizes contrast and precision without requiring manual calibration, balancing ease of operation with measurement accuracy
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
This approach effectively compensates for the meniscus effect, maintaining high contrast and image quality by ensuring the direct radiation is properly aligned with the phase-shifting element, even in microtiter plates with varying well diameters and object liquids, thereby enhancing the visibility of specimens.
Implementation Method 1
an object liquid meniscus forms, which typically has a profile of the surface of the object liquid that deviates from a right angle with respect to the direction of gravity and thus the main radiation direction of the illumination beam, in particular at the periphery of the specimen vessel, and thus accordingly refracts the illumination beam
Implementation Method 2
A component, which has not been diffracted by the specimen, of the illumination beam, which is the main maximum in the original propagation direction of the radiation and will be referred to as direct radiation below, is phase-shifted and attenuated using a phase-shifting element, comprising a phase-shifting retardation element and a gray filter
Implementation Method 3
Due to interference of the direct radiation, which has been manipulated using the phase-shifting element, with the specimen radiation, the resulting phase shift between direct radiation and specimen radiation is converted into an amplitude change
Data Source
AI summary
A method for phase-contrast microscopy on a specimen, and the microscope and optical unit for carrying out the method, is provided. The specimen is arranged in a vessel having an object liquid, in which a component of an illumination beam, which serves as direct radiation and has not been diffracted by the sample, is applied to a phase-shifting element, is characterized in that the illumination beam is deflected before it enters the object liquid by way of an optical deflection element which is changeable in terms of position and/or shape so as to compensate for a change in the main radiation direction of the illumination beam that occurs due to optical refraction at the object liquid.