Tissue-Section Nucleic Acid Testing With Spatial Detection
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Solution Overview
Problem
Conventional nucleic acid testing devices struggle to amplify nucleic acids while retaining position information and perform real-time detection, limiting the acquisition of information such as expression level and position of target nucleic acids.
Innovation Solution
A nucleic acid testing device with a stage, temperature adjuster, intensity detector, and storage unit that allows for real-time nucleic acid amplification and detection, along with a method involving temperature adjustment, label intensity detection, and information storage.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a real-time PCR device is used to amplify nucleic acid, then the amplification can be detected in real time, but position information of the nucleic acid is lost due to tissue crushing and mixing
Solution Approach 1:
The tissue is sectioned into thin slices (5-10 μm thickness) and placed on a stage, maintaining spatial segmentation of different tissue regions. This allows real-time PCR amplification to occur in situ within each section, preserving position information while enabling real-time detection through the transparent section structure.
Solution Approach 2:
A transparent cover glass is introduced as an intermediary between the tissue section and the detection system. This cover glass maintains close proximity to the tissue section for efficient signal detection while allowing optical access for real-time monitoring of amplification reactions without disrupting tissue position.
2Loss of information
If an in situ PCR device is used to retain position information of nucleic acid, then position information can be acquired, but real-time detection and quantitative analysis cannot be performed
Solution Approach 1:
The system transitions from static in situ PCR to dynamic real-time monitoring by implementing continuous temperature cycling and sequential optical detection. The temperature adjuster dynamically controls heating and cooling cycles, while the detector continuously monitors fluorescence signals at each cycle, enabling both position retention and real-time quantitative analysis.
Solution Approach 2:
The amplification process is made continuous through real-time monitoring at each temperature cycle rather than single endpoint detection. The system continuously adjusts temperature through multiple cycles (denaturation, annealing, extension) and continuously detects fluorescence signals, maintaining uninterrupted useful action for both amplification and detection.
3Productivity
If tissue is crushed and mixed for nucleic acid extraction, then amplification can occur, but position information and spatial distribution are lost
Solution Approach 1:
Tissue sections are prepared in advance with optimal thickness (5-10 μm) and mounted on stages before amplification. This preliminary preparation maintains spatial structure intact, allowing subsequent amplification to proceed in situ without requiring crushing or mixing, thus preserving spatial distribution information while ensuring sufficient nucleic acid accessibility.
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 detection of nucleic acid amplification while retaining position information, allowing for the acquisition of more detailed information about the expression level and position of target nucleic acids.
Implementation Method 1
a temperature adjuster that adjusts the temperature of the tissue section on the stage
Implementation Method 2
an intensity detector that detects label intensity in the tissue section over time
Data Source
AI summary
A nucleic acid testing device includes: a stage on which is placed a tissue section to which a solution has been added, in which the solution contains a labeling substance of a target nucleic acid and an amplification reagent for the target nucleic acid; a temperature adjuster that adjusts the temperature of the tissue section on the stage; a temperature controller that controls the temperature adjuster to advance nucleic acid amplification reaction in the tissue section; an intensity detector that detects label intensity in the tissue section over time; and a storage unit that stores detection information generated by the intensity detector.