Calcium Ion Removal for Rapid Fluorescence Detection
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Solution Overview
Problem
Existing methods for detecting cancer cells using fluorescent probes struggle to differentiate between surface and internal cancer cells, as internal cells require time to emit fluorescence, making it difficult to detect cancer cells present within cell masses or tissues.
Innovation Solution
Removing calcium ions from the cell mass or tissue using a buffer solution with a metal ion chelating agent allows target cells to rapidly emit fluorescence when contacted with a fluorescent probe that reacts with enzymes or iron ions, enabling effective detection of internal cancer cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Speed
If a fluorescent probe is directly applied to a tumor region, then the probe can contact the target cells, but internal cancer cells cannot rapidly emit fluorescence
Solution Approach 1:
The patent applies a calcium ion removal solution to the tissue sample before applying the fluorescent probe. This preliminary treatment removes calcium ions that would otherwise bind to the probe and prevent it from reaching and reacting with target cells, especially internal cancer cells. By performing this preparation step in advance, the probe can then rapidly emit fluorescence upon contact with target cells throughout the tissue.
2Measurement precision
If calcium ions are present in the tissue, then the tissue maintains its natural state, but the fluorescent probe cannot effectively detect internal target cells
Solution Approach 1:
The patent changes the chemical environment of the tissue by removing calcium ions using a chelating agent. This parameter change (removal of calcium ions) enables the fluorescent probe to function effectively by preventing calcium-probe binding that would otherwise block probe access to target cells. The method maintains versatility across different tissue types by using a general calcium removal approach that can be applied to various adherent cell types.
3Illumination intensity
If the fluorescent probe is used without pretreatment, then the procedure is simple, but fluorescence intensity is insufficient and rapid disappearance occurs
Solution Approach 1:
The patent introduces a preliminary calcium ion removal step using a buffer solution containing a metal ion chelating agent (such as EDTA or EGTA) before applying the fluorescent probe. This pretreatment prevents calcium ions from interfering with probe binding and fluorescence emission, resulting in significantly enhanced and sustained fluorescence intensity. The additional step, while increasing procedure complexity, is essential for achieving reliable detection of internal cancer cells.
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 allows for the rapid emission of fluorescence from internal target cells, enhancing the ability to detect cancer cells within tissues, improving diagnostic accuracy and enabling reliable removal during surgery.
Implementation Method 1
removing calcium ions from within a cell mass or a tissue containing the target cell by impregnating the cell mass or the tissue with a buffer solution containing a metal ion chelating agent
Implementation Method 2
contacting the cell mass or the tissue with a fluorescent probe which emits fluorescence when contacted with or taken up into the target cell, wherein the fluorescent probe emits fluorescence when reacted with an enzyme or iron ions contained in the target cell
Data Source
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AI summary
The purpose of the present invention is to provide a method for allowing a target cell to emit light, in which the target cell is present inside of a cell mass and is aimed to emit light by a fluorescent probe. The present invention provides a method for allowing the target cell to emit fluorescence comprising removing calcium ions from within the cell mass or a tissue containing the target cell and contacting the cell mass or the tissue with the fluorescent probe which emits fluorescence when contacted with or taken up into the target cell, thereby allowing the target cell present inside of the cell mass or the tissue to emit light, and thus can be used in a detection of the target cell in a sample, such as a clinical sample, in which the target cell may not be present on the surface of the sample.