Pressure-Driven Fluorescence Measurement for Tadpoles
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Solution Overview
Problem
Existing methods for fluorescence measurements and sorting of large multi-cellular organisms in a flowing liquid often cause physical stress and damage to the organisms, particularly when using mechanical pumps, which are not suitable for soft particles like tadpoles.
Innovation Solution
A system utilizing air pressure or gravity to create a pressure differential for non-destructive pumping of organisms through a fluorescence measuring device, combined with LED illumination and digital imaging for real-time fluorescence measurement, and an optional sorting mechanism based on fluorescence intensity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If mechanical pumps are used to pump organisms through the measurement device, then the flow rate and measurement speed are improved, but the organisms suffer physical stress and damage
Solution Approach 1:
The patent replaces mechanical pumps with a pressure-driven flow system that uses a reservoir and pressure differential to move organisms through the measurement chamber, eliminating mechanical contact and stress on the organisms while maintaining continuous flow capability
Solution Approach 2:
The system uses hydraulic pressure from a fluid reservoir to drive the flow of organisms through the measurement device, providing a non-mechanical pumping action that is gentle on biological samples while achieving the required flow rates for real-time measurement
2Measurement precision
If traditional flow cytometry methods are used, then the fluorescence measurement capability is achieved, but the method is not suitable for large multi-cellular organisms
Solution Approach 1:
The patent transitions from point-detector flow cytometry to an imaging-based system that captures two-dimensional images of entire organisms, allowing fluorescence measurement across the full body of large multi-cellular organisms rather than just passing them through a narrow detection point
Solution Approach 2:
The system is designed to handle a wide range of organism sizes and types by using a spacious measurement chamber and flexible imaging approach, making it universally applicable to both small cells and large multi-cellular organisms like Xenopus tadpoles
3Reliability
If measurements are performed on living organisms, then the physiological relevance is improved, but the organisms may be harmed during the process
Solution Approach 1:
The system uses gentle pressure-driven flow and optimized imaging parameters to prevent damage before it occurs, ensuring organisms remain viable throughout the measurement process and allowing repeated measurements on the same individuals
Solution Approach 2:
The organisms remain in their natural aquatic environment throughout the measurement process, maintaining their physiological state without requiring extraction or special handling that could cause stress, thereby preserving the physiological relevance of the fluorescence measurements
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 real-time fluorescence measurement and sorting of large multi-cellular organisms with minimal physical stress, allowing for repeated measurements without harming the organisms, and is particularly effective for Xenopus tadpoles, providing a sensitive and specific testing method for pollutants and pharmaceutical applications.
Implementation Method 1
A sample reservoir (402) and air pipe (404) are provided. A pump (412) pumps air (602) through the air pipe (404) into the sample reservoir (402). The pressure differential moves the sample through a fluorescence-measuring device
Implementation Method 2
A first cylinder (806) and a second cylinder (826) are provided in this gravity embodiment. The gravity creates a pressure differential that moves the sample through the flow cell
Implementation Method 3
The preferred technology for measuring organism fluorescence, presented here, is the fountain flow cytometry technique summarized above and described in detail in U.S. Pat. No. 6,765,656, combined with LED illumination and a digital imager
Implementation Method 4
Xenopus tadpoles 'light up' (exhibit fluorescence) in response to a pollutant (or drug)
Data Source
AI summary
Apparatus and methods for measuring the fluorescence of large multi-cellular organisms in a sample of liquid includes a pumping mechanism, a fluorescence measuring device, a method of analyzing the measurements, and optionally, a sorting mechanism. The pumping mechanism transfers large multi-cellular organisms from a reservoir through a fluorescence-measuring device causing minimum physical damage and/or stress. The pressure differential driving the organisms from a sample container/reservoir through the measuring device can be derived from gravity, air pressure, or liquid pressure, or some combination of the three. The fluorescence can be measured in a cytometer using a light detector or imager. Generally the detection element will include a filter, isolating the wavelength of fluorescent emission. The illumination may be provided by a laser or by an LED, combined with the use of dichroic mirrors to allow multiple wavelength simultaneous illumination.