Microorganism Detection via Fluorescence Staining and Dilution
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Solution Overview
Problem
Existing methods for examining microorganisms in ballast water, such as those described in Patent Literatures 1 and 2, are large-scale and costly, with high production costs and long measurement times, and suffer from reduced detection accuracy due to high background fluorescence interference.
Innovation Solution
A method involving a sampling preparation step that includes stirring and mixing a sample with a fluorescence staining reagent, followed by a still standing period and dilution with a non-fluorescent liquid, and optionally a pH adjustment step, to enhance the clarity of microorganism staining and reduce background fluorescence, thereby improving the signal-to-noise ratio and detection accuracy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of time
If seawater is pumped and sequentially passed through staining, concentrating, and measurement steps in a flow system, then waiting time is reduced and stable staining information is acquired, but the apparatus becomes large-scale and high in production cost
Solution Approach 1:
The patent combines multiple functions (staining, concentrating, and measurement) into a single integrated flow system apparatus. The water pump, flow cells, and measurement components are merged into one cohesive system that processes seawater continuously, eliminating the need for separate batch processing steps and reducing overall apparatus complexity while maintaining functional integration.
Solution Approach 2:
The staining process is performed preliminarily in the flow system before measurement, with the water pump continuously circulating seawater through the staining portion and then to the measurement portion. This preliminary staining action in flow mode eliminates waiting time between staining and measurement, as the process continues uninterrupted through the integrated system.
2Reliability
If seawater is sequentially passed through staining and measurement steps, then stable staining information is acquired, but measurement completion takes at least several hours
Solution Approach 1:
The patent implements continuous circulation of seawater through the flow system, where the water pump continuously moves water from the staining portion through the flow cells to the measurement portion without interruption. This continuous action maintains stable staining conditions throughout the process while significantly reducing total measurement time compared to batch methods that require waiting between steps.
Solution Approach 2:
The patent replaces manual batch processing mechanics with an automated flow system driven by a water pump. The mechanical circulation system continuously transports seawater through staining and measurement stages, eliminating manual intervention and waiting periods while maintaining staining stability through controlled continuous flow conditions.
3Measurement precision
If fluorescence staining reagent is added to sample for microorganism detection, then microorganism detection capability is enhanced, but background fluorescence increases and reduces detection accuracy
Solution Approach 1:
The patent extracts and removes background fluorescence components from the sample before performing fluorescence staining and measurement. By pre-processing the seawater sample to eliminate interfering background fluorescence, the system enhances the signal-to-noise ratio and improves detection accuracy of microorganisms, allowing the fluorescence staining reagent to work more effectively without interference.
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 allows for more accurate and efficient detection of microorganisms by significantly reducing background fluorescence, leading to a substantial enhancement in detection accuracy and reducing the time required for measurements.
Implementation Method 1
a fluorescence staining step of stirring and mixing certain amounts of the sample and the fluorescence staining reagent
Data Source
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AI summary
A method for examining microorganisms has a sampling preparation step including a fluorescence staining step of stirring and mixing certain amounts of a sample and a fluorescence staining reagent, a still standing step of leaving the solution after the fluorescence staining step to still stand for a certain time, and a dilution step of diluting the solution after the still standing step with a liquid that emits no fluorescence.