Peltier Thermal Cycling for Rapid Cell Lysis in Closed Tubes
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Solution Overview
Problem
Current cell disruption methods are time-consuming, damaging to nucleic acid, prone to contamination, and not amenable to automation, particularly when extracting nucleic acid from complex organisms like bacteria, which often require additional steps and toxic chemicals.
Innovation Solution
A thermoelectric cell-based process that rapidly disrupts cells through controlled thermal cycling between freezing and boiling temperatures, using a Peltier cell with a heat source/sink and optional resistive heating to optimize temperature and minimize thermal stress, allowing for efficient release of nucleic acid components without the need for toxic additives.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If conventional cell disruption methods (acid/base, enzymatic, sonication, mechanical separation) are used, then cell membranes are disrupted and nucleic acid is released, but the process is time-consuming and damaging to nucleic acid quality
Solution Approach 1:
The patent applies phase transitions by rapidly cycling the temperature between freezing and boiling points. This thermal cycling causes repeated expansion and contraction of cellular structures, leading to membrane disruption and nucleic acid release. The phase changes of water (liquid ↔ solid ↔ gas) provide the mechanical force needed for cell lysis without requiring prolonged processing or harsh chemicals.
Solution Approach 2:
The invention employs periodic thermal action by alternating between heating and cooling cycles. This periodic temperature manipulation creates repeated stress on cell membranes, progressively disrupting them over multiple cycles. The rhythmic application of thermal energy achieves complete cell lysis faster than continuous methods, reducing overall process time while preserving nucleic acid integrity.
2Quantity of substance
If conventional cell disruption methods are used, then cell membranes are disrupted, but the process is prone to contamination and not amenable to automation
Solution Approach 1:
The patent replaces manual mechanical disruption methods (sonication, mechanical separation) with an automated thermal cycling system. The temperature-controlled environment automatically performs heating and cooling cycles without requiring manual intervention for each sample. This substitution enables high-throughput processing and automation while maintaining consistent results across multiple samples, eliminating contamination risks associated with manual operations.
3Quantity of substance
If toxic chemicals (phenol, guanidium salts) are used for DNA extraction, then unwanted cellular components are removed, but dangerous chemicals are introduced into the process
Solution Approach 1:
The invention converts the potentially harmful effect of extreme temperature changes into a beneficial purification mechanism. By using controlled thermal cycling, the method achieves cell lysis and nucleic acid release without requiring toxic chemicals. The thermal stress selectively disrupts cellular membranes and denatures proteins while leaving nucleic acids intact, thereby purifying the target molecule without introducing harmful substances.
Solution Approach 2:
The patent employs a disposable thermocycler tip or reaction vessel that can be discarded after a single use. This eliminates the need for extensive cleaning and decontamination steps required when using toxic chemicals. The single-use nature of the container ensures that no residual chemicals contaminate subsequent samples, providing inherent purification while avoiding toxic exposure.
4Productivity
If direct amplification from complex samples (bacterial cells) is attempted, then nucleic acid can be obtained rapidly, but additional steps and toxic additives are required
Solution Approach 1:
The patent merges the cell lysis step and nucleic acid release step into a single thermal cycling process. By combining multiple functions (membrane disruption, protein denaturation, nucleic acid liberation) into one automated temperature cycling protocol, the method eliminates the need for separate processing steps and toxic additives. This integration achieves rapid direct amplification from complex samples while simplifying the overall workflow to a single操作步骤.
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 method enables rapid, efficient, and controlled cell disruption suitable for a wide range of organisms, allowing for direct amplification of DNA from complex samples in a single closed tube, reducing the need for intermediate steps and minimizing contamination, and is amenable to automation.
Implementation Method 1
applying electric current to the thermoelectric cell to substantially change the temperature of the material
Implementation Method 2
the base face of which is in close proximity to a heat source/sink at a substantially constant temperature between the freezing and boiling temperatures of water
Implementation Method 3
The process of amplifying nucleic acid sequences directly from simple targets such as human blood has been previously described. The cell types in these previous works are highly susceptible to lysis
Data Source
AI summary
Disrupting a biological cell includes freezing, boiling or perhaps alternately freezing and boiling material containing the biological cell using a thermoelectric cell with a working face, and a base face whereof is contiguous with a heat source/sink at a substantially constant temperature. Apparatus for the disruption process includes a peltier cell, a base face, which is flexibly attached to a heat source/sink held at a constant temperature, and a working face contiguous with a reaction vessel or holder thereof. Reversal of the voltage in the peltier cell enables the working face alternately to reach below freezing and above boiling temperatures, and/or with use of a resistive wire on the vessel or holder for heating, with the TEC used purely for cooling. The materials of the base face tend to inhibit disintegration of the peltier cell brought about by expansion and contraction by heat.