Exosome Fluorescence Measurement with Multi-Point Laser Exposure Control
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Solution Overview
Problem
Existing devices for determining fluorescence and antibody number on exosomes cannot perform multiple measurements on the same sample due to excessive bleaching of fluorescent dye caused by prolonged laser exposure, necessitating sample replacement after each measurement.
Innovation Solution
A device with five lasers, a multi-notch filter, and a quartz glass to maintain consistent optical path lengths, combined with a video camera using a graphene-based light sensor and fast stepper motors, allows for rapid repositioning and sequential measurement at multiple points within the measuring cell, minimizing laser exposure time to preserve fluorescence.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If laser exposure time is prolonged to obtain sufficient measurement data, then measurement precision is improved, but fluorescent dye bleaching increases causing loss of fluorescence signal
Solution Approach 1:
The patent divides the measurement process into multiple discrete measurement points distributed throughout the sample volume. Instead of continuous exposure at one location, the system sequentially visits multiple positions (e.g., 100 measurement points) to accumulate statistical data, thereby reducing cumulative exposure time at any single point while maintaining measurement precision through statistical aggregation.
Solution Approach 2:
The patent transitions from temporal accumulation (prolonged exposure at one point) to spatial distribution (multiple points in 3D space). By measuring at multiple distinct locations within the sample cell and aggregating results, the system achieves sufficient statistical data without excessive local exposure, effectively using the spatial dimension to resolve the time-exposure dilemma.
2Reliability
If laser exposure time is shortened to reduce dye bleaching, then fluorescence preservation is improved, but measurement precision deteriorates due to insufficient data accumulation
Solution Approach 1:
The patent segments the measurement task across multiple spatial locations rather than concentrating it in time at a single location. Each brief exposure at a measurement point contributes to the overall statistical dataset, and by aggregating data from many such points, sufficient precision is achieved without requiring prolonged exposure at any single point.
Solution Approach 2:
The patent effectively creates multiple copies of the measurement process at different spatial locations. Instead of one prolonged measurement, the system performs many brief measurements at different points, where each measurement is a copy of the basic measurement process. The statistical combination of these copies provides the necessary precision while preserving fluorescence.
3Reliability
If multiple measurements are performed on the same sample, then statistical reliability is improved, but sample replacement is required after each measurement due to dye bleaching
Solution Approach 1:
The patent segments the sample measurement into multiple spatial locations within the same sample cell. By distributing measurement points throughout the sample volume, the system enables multiple measurements on the same sample without depletion or bleaching issues, eliminating the need for sample replacement while maintaining statistical reliability.
Solution Approach 2:
The patent uses the spatial dimension to enable repeated measurements. Instead of measuring the same location repeatedly (which causes bleaching), the system measures at multiple distinct spatial positions, allowing unlimited measurements on the same sample as long as the sample volume is sufficient to contain all measurement points.
4Measurement precision
If laser beam focusing is optimized to improve measurement precision, then measurement quality is improved, but laser exposure time must be reduced to prevent dye bleaching
Solution Approach 1:
The patent segments the laser exposure into many brief instances at different locations rather than one prolonged exposure. By distributing the total exposure time across multiple spatial points, the system can use optimized focusing for high precision at each point while the cumulative exposure time remains low enough to prevent bleaching.
Solution Approach 2:
The patent employs periodic sequential measurement at different locations. The laser visits measurement points in a systematic sequence, spending brief periods at each location. This periodic action allows optimized focusing and maximum measurement quality at each point while the periodic nature ensures total exposure time remains controlled to prevent bleaching.
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 up to 100 measurements on a single exosome sample with preserved fluorescence, ensuring high measurement quality and statistical reliability by protecting the dye from bleaching.
Implementation Method 1
determining the fluorescence and number of antibodies on exosomes
Implementation Method 2
the focusing of the laser beam (21) in interaction with the sample (9) forms the center of a convergent beam bundle, consisting of light from the fluorescence plane (5) and the scattering plane (8)
Data Source
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AI summary
The invention relates to a device and method for reducing the reduction in intensity of a fluorescence dye by laser light when determining fluorescence and the number of antibodies on exosomes, comprising means for storing different measurement points of various differently coloured lasers in a measuring cell at certain measurement positions, the focussing of the laser beam interacting with the sample being recorded in a video camera as the centre of a convergent beam bundle.