Resin Particle Internal Standard for Blood Cell Quantification
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Solution Overview
Problem
Current methods for detecting rare cells in blood samples, such as circulating tumor cells, face challenges in reliability due to variations in separation media density, cell loss, and inaccurate calibration, leading to errors in cell counting.
Innovation Solution
A method involving density gradient centrifugation with resin particles of specific gravities matching those of rare cells, allowing for accurate separation and quantification by using resin particles as internal standards to evaluate system reliability and correct cell counts.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If density gradient centrifugation is used to separate rare cells from blood samples, then cell separation is achieved, but reliability decreases due to variations in separation media density and inaccurate calibration
Solution Approach 1:
Resin particles with specific gravities matching rare cells serve as intermediary reference objects during density gradient centrifugation. These particles co-separate with rare cells through the density gradient, enabling calibration of separation efficiency and detection reliability without interfering with the actual cell separation process
Solution Approach 2:
The invention introduces resin particles with specifically controlled specific gravities (matching rare cell densities) as reference standards. By varying the specific gravity parameter of the reference particles to match different cell types, the system achieves accurate calibration for different separation conditions and media compositions
2Reliability
If stabilized cells are used as internal controls, then detection reliability improves, but separation effectiveness deteriorates due to specific gravity differences
Solution Approach 1:
The invention changes the key parameter of the reference objects from biological cells to resin particles with tunable specific gravities. This allows precise matching of the reference particle density to the rare cells being detected, enabling both reliable detection and effective separation simultaneously
Solution Approach 2:
Instead of using stabilized cells that copy biological properties, the invention uses resin particles that copy the physical property (specific gravity) of rare cells. This simplified model achieves the calibration function without the complicating factor of biological variability
3Ease of operation
If separation media density varies with ambient environment, then ease of operation is maintained, but measurement precision deteriorates
Solution Approach 1:
The resin particles serve as internal reference standards that provide feedback on the actual separation performance under varying environmental conditions. By comparing the position and distribution of reference particles with expected values, the system can detect and correct for density variations in the separation media
Solution Approach 2:
The resin particles act as intermediary reference objects that mediate between the variable separation conditions and the detection system. They translate environmental variations into measurable positional changes that can be used to calibrate and correct the detection results
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 enhances the reliability of cell quantification by minimizing errors in density gradient centrifugation and providing a calibrated system for precise counting of rare cells, improving the accuracy of clinical tests.
Implementation Method 1
Separating a blood-derived sample containing a known number (P0) of resin particles P having a specific gravity larger than 1.025 but smaller than 1.120 and a known number (N0) of resin particles N having a specific gravity of not less than 1.090 and not more than 1.120 by density gradient centrifugation
Data Source
Figure 1
AI summary
An object of the present invention is to provide a method for quantifying cells of interest potentially contained in a blood-derived sample in cases of their quantification after separation from the blood-derived sample, which method enables accurate quantification of the cells without causing underestimation of the cell number due to loss of the cells of interest during the process from immediately after blood collection to the separation or causing erroneous quantification of the cell number due to inaccurate calibration of the apparatus for detecting the cells of interest. The quantification method of the present invention is a method for quantifying cells of interest having a specific gravity larger than a specific gravity of blood plasma but smaller than a specific gravity of erythrocytes, the method includes the steps of : (A) separating a blood-derived sample containing a known number (P0) of resin particles P having a specific gravity larger than a specific gravity of blood plasma but smaller than a specific gravity of erythrocytes by density gradient centrifugation into at least two layers including a layer of erythrocytes and a layer of cells other than erythrocytes; (B) extracting the layer of cells other than erythrocytes and counting the number of cells of interest and the number (P1) of the resin particles therein; and (C) correcting the number of cells of interest by multiplying the number of cells of interest by P0/P1.