Paraffin Removal by Phase Separation for Biomolecule Isolation
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Solution Overview
Problem
Current methods for isolating biomolecules from fixed, paraffin-embedded biological samples are complex, time-consuming, and prone to material loss due to the need for multiple washing steps and solvent removal, making automation difficult, especially with small sample amounts.
Innovation Solution
A method involving contact with a water-immiscible solvent to dissolve paraffin, forming a liquid organic phase that remains in the sample container, allowing biomolecules to be isolated from an aqueous phase without the need for solvent removal or extensive washing, enabling automated processing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If multiple washing steps with ethanol are performed to remove paraffin and solvent, then the sample material becomes cleaner, but the process becomes time-consuming and sample material is lost
Solution Approach 1:
The invention extracts and removes the paraffin solvent system entirely by using a water-immiscible solvent that forms a separable organic phase. The paraffin is dissolved in this solvent and then the entire organic phase is removed in one step, eliminating the need for multiple ethanol washing steps that were previously required to achieve clean sample material.
Solution Approach 2:
The invention changes the solvent parameter from water-miscible (ethanol) to water-immiscible (such as chloroform, dichloromethane, or hexane). This parameter change allows the solvent and dissolved paraffin to form a separate organic phase that can be easily removed, eliminating the need for multiple washing steps and reducing both time and sample material loss.
2Manufacturing precision
If multiple washing steps and centrifugation are performed to remove paraffin, then the sample material becomes cleaner, but the complexity of the procedure increases and automation becomes difficult
Solution Approach 1:
The invention extracts the paraffin into a water-immiscible solvent forming a distinct organic phase, which is then removed in a single step. This extraction approach simplifies the procedure by eliminating multiple washing and centrifugation steps, making the process suitable for automation while maintaining sample cleanliness.
Solution Approach 2:
Instead of using water-miscible solvents that require sequential washing to remove, the invention uses water-immiscible solvents that naturally separate into an organic phase. This inversion of the solvent selection approach reverses the typical workflow, allowing for simpler single-step removal rather than multiple washing steps.
3Manufacturing precision
If the organic solvent is removed by evaporation or centrifugation, then the sample is cleaned, but sample material is lost especially when small amounts are available
Solution Approach 1:
The invention extracts paraffin into a water-immiscible solvent and removes the entire organic phase containing the dissolved paraffin in one step. This approach avoids the need for evaporation or repeated centrifugation that could lose small amounts of sample material, as the removal is achieved through simple phase separation and decanting.
Solution Approach 2:
The water-immiscible solvent acts as an intermediary that selectively dissolves paraffin but not the biomolecule-containing aqueous phase. This intermediary allows for clean separation and removal of paraffin without affecting the sample material, preventing loss that would occur with direct evaporation or centrifugation of the sample itself.
4Manufacturing precision
If heating above the melting point of paraffin is performed to dissolve it, then the paraffin becomes liquid and can be removed, but the sample material may be damaged and the process becomes complex
Solution Approach 1:
The water-immiscible solvent serves as an intermediary that dissolves paraffin at ambient temperatures without requiring heating. This intermediary approach avoids the harmful thermal effects on the sample material while still achieving effective paraffin dissolution and removal through the solvent's ability to solubilize paraffin at room temperature.
Solution Approach 2:
The invention changes the temperature parameter from elevated (heating above melting point) to ambient temperature. By using a water-immiscible solvent with high paraffin solubility at room temperature, the process eliminates the need for heating, thereby preventing thermal damage to the sample material while maintaining effective paraffin removal.
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 significantly reduces process effort, minimizes sample loss, and facilitates automated sample processing by maintaining a liquid organic phase at ambient temperatures, allowing for efficient isolation of various biomolecules like nucleic acids and proteins.
Implementation Method 1
Bringing the sample into contact a) with a water-immiscible solvent and dissolving the paraffin in this solvent to form a liquid organic phase
Implementation Method 2
contact with an aqueous solution of at least one lysis substance and/or with water and at least one lysis substance to form an aqueous phase
Data Source
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AI summary
The present invention relates to a method for isolating biomolecules from a fixed, paraffin-embedded biological sample, comprising the steps of: contacting the sample a) with an immiscible solvent and dissolving the paraffin in this solvent to form a liquid organic phase, and b) with an aqueous solution of at least one lysis agent and/or with water and at least one lysis agent to form an aqueous phase; and isolating the biomolecules from the aqueous phase. The invention also relates to a corresponding kit and the use of this kit for isolating biomolecules.