Microscope Stage Mount with Built-in Fiduciary Markers
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Solution Overview
Problem
Current high-resolution fluorescence microscopy techniques require expensive and complex equipment to stabilize the microscope stage, making them difficult to use effectively and prone to sample interference, while simpler methods using gold particles have limitations in placement and biocompatibility.
Innovation Solution
A cost-effective, easy-to-make microscope stage mount with a stably built-in ordered two-dimensional array of fiduciary markers made of poly(methyl methacrylate) (PMMA) or polymethylglutarimide (PMGI) pillars, which are optically transparent, biocompatible, and easily detectable using infrared light for precise calibration.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If expensive and complex equipment is used to stabilize the microscope stage, then stabilization precision is improved, but device complexity and cost increase
Solution Approach 1:
The patent replaces expensive, complex stabilization equipment with inexpensive fiducial markers (fluorescent beads or fluorescently labeled antibodies) that are easily applied and removed. These markers provide stable reference points for drift correction without requiring sophisticated hardware, thereby achieving high stabilization precision while minimizing device complexity and cost.
Solution Approach 2:
The patent introduces fiducial markers as intermediary reference objects between the microscope stage and the imaging system. These markers serve as mediators that enable precise measurement and correction of stage drift through software-based tracking, eliminating the need for complex mechanical stabilization systems while maintaining nanometer-level precision.
2Difficulty of detecting and measuring
If gold particles or beads are used as fiducial markers, then marker detectability is improved, but ease of manufacture and biocompatibility worsen
Solution Approach 1:
The patent employs commercially available fluorescent beads or fluorescently labeled antibodies as disposable fiducial markers. These markers can be easily applied to the sample using standard microscopy techniques, require no special manufacturing equipment, and can be removed or replaced between experiments, thereby greatly simplifying the manufacturing and placement process while maintaining excellent detectability.
Solution Approach 2:
The patent utilizes fluorescent markers that emit light at wavelengths optimized for detection by standard fluorescence microscopes. By selecting markers with appropriate fluorescence properties (emission wavelength, brightness, photostability), the system achieves high detectability using conventional equipment, eliminating the need for complex detection systems while improving ease of manufacture and use.
3Measurement precision
If fiducial markers are added to correct stage drift, then measurement precision is improved, but device complexity increases
Solution Approach 1:
The patent replaces mechanical stage stabilization systems with a software-based drift correction system that tracks fiducial markers in the image sequence. This computational approach eliminates the need for complex mechanical actuators, sensors, and control systems, thereby achieving high drift correction precision while significantly reducing overall system complexity.
Solution Approach 2:
The fiducial markers themselves serve as self-contained reference objects that automatically provide drift information throughout the imaging process. The markers passively emit fluorescence that can be continuously tracked without requiring active control mechanisms, power sources, or complex instrumentation, thereby achieving precise drift correction with minimal added system complexity.
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 efficient and accurate stabilization of the microscope stage to nanometer precision without interfering with biological samples, facilitating high-resolution imaging in both in vitro and living cells, and physiological tissues with minimal equipment requirements.
Implementation Method 1
easily detectable using infrared light for precise calibration
Implementation Method 2
easily detectable using infrared light for precise calibration
Data Source
AI summary
An improved microscope stage mount with built-in fiduciary markers is used for fluorescence microscopy, and comprises: (a) an optically-transparent glass plate adapted for specimen mounting and microscope viewing and comprising a specimen mounting area; and (b) a defined and ordered, two-dimensional microscopic array of fiduciary markers, wherein the markers are polymeric pillars affixed to the plate about the specimen mounting area, wherein the markers provide a three-dimensional spatial reference for the specimen.