TIRF Observation Device Angle Adjustment for Evanescent Light
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Solution Overview
Problem
Existing fluorescence observation methods using evanescent light face challenges with fluctuating excitation light intensity due to substrate surface irregularities, leading to inconsistent results and difficulty in distinguishing between partially and totally reflected light, which affects the reliability and sensitivity of measurements.
Innovation Solution
A method and device that automatically adjust the angle of incidence of excitation light using optical sensors to ensure total reflection on the substrate surface, continuously varying the angle and sensing the state of the light after incidence to maintain optimal total reflection, thereby maintaining high evanescent light intensity and improving operability and reliability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Illumination intensity
If manual angle adjustment is used to achieve total reflection, then excitation light intensity can be optimized for flat substrates, but angle of incidence varies due to substrate surface irregularities causing fluctuating excitation light intensity
Solution Approach 1:
The patent employs optical sensors to detect the angle of incidence in real-time and feeds this information back to an angle adjusting mechanism. This closed-loop feedback system continuously monitors and corrects angle variations caused by substrate surface irregularities, maintaining consistent total reflection conditions and stable excitation light intensity across the substrate surface.
Solution Approach 2:
The patent transitions from static manual angle adjustment to dynamic automatic angle adjustment. The angle of incidence is continuously varied and adjusted based on real-time detection, allowing the system to adapt to substrate surface variations and maintain optimal total reflection conditions throughout the observation process.
2Ease of operation
If a fixed angle of incidence is set, then optical adjustment is simplified, but it is difficult to distinguish between partially and totally reflected light affecting measurement reliability
Solution Approach 1:
Optical sensors provide real-time feedback on the reflection state by detecting the angle of incidence and the characteristics of reflected light. This feedback enables the system to automatically determine whether total reflection is occurring and adjust accordingly, resolving the ambiguity between partial and total reflection states.
Solution Approach 2:
The patent replaces manual mechanical angle adjustment with an automated optical detection and control system. Optical sensors detect the reflection state and trigger automatic angle adjustment, substituting human judgment and manual manipulation with instrumental measurement and automated control for more precise distinction between reflection types.
3Reliability
If angle of incidence is increased above critical angle to ensure total reflection, then total reflection is achieved, but depth of evanescent light oozing decreases reducing signal intensity
Solution Approach 1:
The system dynamically adjusts the angle of incidence within the critical angle range rather than using a fixed angle above the critical angle. By continuously optimizing the angle to maintain total reflection while staying as close as possible to the critical angle, the system maximizes evanescent light penetration depth and intensity while ensuring reliable total reflection conditions.
Solution Approach 2:
The patent optimizes the angle of incidence parameter by making it variable and adjustable based on real-time conditions. Instead of using a fixed angle significantly above the critical angle, the system adjusts the angle parameter dynamically to achieve the optimal balance between ensuring total reflection and maximizing evanescent light intensity for fluorescence excitation.
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 enhances the operability and reliability of fluorescence observation by maintaining consistent excitation light intensity and reducing the impact of substrate surface irregularities, resulting in higher signal intensity and improved measurement accuracy.
Implementation Method 1
When light having an angle equal to or larger than a fixed angle is made incident from a medium having a high refractive index to a medium having a low refractive index, the incident light does not diffuse to the medium having the low refractive index and total reflection occurs
Implementation Method 2
total reflection occurs. At this point, a phenomenon in which the light slightly oozes out occurs on a surface on the low-refractive index medium side of a boundary surface
Implementation Method 3
The oozing-out light is called evanescent light. The intensity of the evanescent light is attenuated exponentially further away from a refractive index boundary plane
Implementation Method 4
When a living organism substance such as DNA or protein is observed, it is a general practice to use a method of performing marking with a fluorescent dye, irradiating excitation light such as a laser, and observing generated fluorescent light
Data Source
AI summary
A device and method for fluorescence observation have good operability, high sensitivity, and high acid reliability. The device is used for fluorescence observation using evanescent light. The angle of incidence of the excitation light is adjusted so that the excitation light is totally reflected from the surface of a substrate irrespective of the angle of the substrate surface. The method includes a step of shining the excitation light on the observation substrate while continuously varying the angle of the excitation light with respect to the observation substrate. In addition, the method includes a step of sensing the shone excitation light via optical sensors, and a step of setting the angle of total reflection according to the result of the sensing by the optical sensors. In the present device and method, the direction in which the shone excitation light travels varies with the angle of incidence.