Fluorescence Microscope Calibration Apparatus for Excitation Power Measurement
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Solution Overview
Problem
Fluorescence microscopy faces challenges in comparing experimental results due to differences in detection characteristics between microscopes and over time, and there is a need to minimize phototoxicity by quantitatively estimating excitation power.
Innovation Solution
A calibration apparatus for fluorescence microscopes and detection devices that measures excitation characteristics, including power and wavelength of excitation light, and extracts detector characteristics to generate noise models, improving data analysis and reducing ambiguity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If different microscopes or detection instruments are used to collect experimental data, then various detection characteristics enable flexible measurement capabilities, but differences in detection limits make it difficult to compare experimental results between laboratories or over time
Solution Approach 1:
The patent applies parameter changes by measuring and reporting key parameters of the excitation light (power, wavelength) and detector characteristics (gain, read noise, dark current) to enable normalization and comparison of experimental results across different microscopes and detection instruments. This allows researchers to adjust and compare data based on these measured parameters rather than being constrained by instrument-specific detection characteristics.
2Illumination intensity
If excitation light is used to illuminate the sample to obtain fluorescence images, then sufficient signal intensity is achieved, but phototoxicity occurs that influences the behavior of the sample under observation
Solution Approach 1:
The patent implements feedback by measuring the actual power and wavelength of the excitation light delivered to the sample and reporting these values alongside the fluorescence images. This feedback mechanism allows researchers to monitor and adjust excitation parameters to minimize phototoxicity while maintaining sufficient signal intensity, enabling quantitative assessment of phototoxicity effects in biological experiments.
3Object-affected harmful factors
If quantitative estimate of excitation power is obtained to minimize phototoxicity, then sample integrity is improved, but additional measurement and calibration procedures increase system complexity
Solution Approach 1:
The patent applies self-service by enabling the microscope system to automatically measure and report its own excitation light parameters (power, wavelength) and detector characteristics. This self-measurement capability eliminates the need for complex external calibration procedures and manual measurements, reducing system complexity while providing quantitative excitation power estimates to minimize phototoxicity.
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 quantitative and reliable data analysis by standardizing experimental results and reducing phototoxicity through accurate measurement and reporting of excitation power, enhancing the reliability and efficiency of fluorescence microscopy.
Implementation Method 1
an optical power sensor configured to produce a power signal representative of a power magnitude of light applied to the optical power sensor
Implementation Method 2
an optical wavelength sensor configured to produce wavelength information associated with the light applied to the optical wavelength sensor
Implementation Method 3
certain chemical moieties or other compounds (e.g., a tryptophan amino acid, certain dyes, or fluorescence proteins) in the sample may emit light (termed 'emission light') in the form of fluorescence
Data Source
AI summary
The described embodiments are directed to a system and methods of calibrating a fluorescence microscope and/or light detection device using a calibrating apparatus. The apparatus may comprise a main body housing, a sensor head, and a microcontroller assembly disposed within the housing. The housing may include an adapter to mechanically couple the housing to a microscope. The sensor head may comprise (i) an optical power sensor to produce a power signal representative of an optical power magnitude of light applied to the optical power sensor, (ii) an optical wavelength sensor configured to produce wavelength information associated with the light applied to the optical wavelength sensor, and (iii) a light source configured to direct light toward a detection device associated with the microscope. The microcontroller assembly may be configured to generate an optical power magnitude value based on the power signal and adjusted according to the wavelength information.