Cell Analyzer Optical Spot Shaping for High-Precision Measurement
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Solution Overview
Problem
Existing flow cytometry methods struggle to measure cells of 20 to 100µm in size, such as epithelial cells from the uterine cervix, with high precision due to limitations in detecting size and nucleus size using scattered light and fluorescent light.
Innovation Solution
A cell analyzer and method that uses a specific optical detector system with a semiconductor laser and PI staining to accurately measure cells by forming a beam spot of 3 to 8µm in the flow direction and 300 to 600µm perpendicular to it, allowing for precise detection of cell and nucleus size through forward and side scattered light, and fluorescent light signals.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional flow cytometry with scattered light detection is used, then white blood cells (10μm) can be measured, but cells of 20 to 100μm size cannot be measured with high precision
Solution Approach 1:
The patent changes the optical parameters by using a semiconductor laser with wavelength 630nm or more (red region) instead of conventional shorter wavelengths, and combines it with propidium iodide staining that emits fluorescent light at 650nm or more. This parameter change in light wavelength and staining methodology enables high-precision measurement of larger cells (20-100μm) by improving the signal-to-noise ratio for cells of this size range.
2Measurement precision
If laser light irradiation and scattered light detection are used, then measurement can be performed, but high-precision measurement of large cells (20 to 100μm) is difficult
Solution Approach 1:
The patent introduces propidium iodide as an intermediary substance that stains the cell nucleus. This fluorescent stain acts as a mediator that absorbs the incident laser light (630nm or more) and emits fluorescent light (650nm or more) specifically from the nucleus region. This intermediary enables high-precision nucleus size measurement and improves detection reliability for epithelial cells by providing a strong, specific signal from the nuclear region.
3Measurement precision
If conventional flow cytometry is used for cervical epithelial cells, then measurement is possible, but high-precision differentiation between normal and abnormal cells is difficult
Solution Approach 1:
The patent utilizes color changes in the optical detection by employing red region laser light (630nm or more) and detecting fluorescent light in the red region (650nm or more) from propidium iodide staining. This color change approach provides high contrast between normal and abnormal cervical epithelial cells, enabling precise differentiation. The specific wavelength selection and fluorescent emission detection create distinct signal characteristics that improve abnormal cell detection precision.
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 high-precision measurement of cells and nuclei, improving the detection of abnormal cells by determining the N/C ratio and accurately distinguishing between normal and cancerous or atypical cells based on size and DNA content.
Implementation Method 1
measuring cells by irradiating a measurement sample which includes cells with light and detecting scattered light and fluorescent light from the measurement sample
Implementation Method 2
detecting scattered light and fluorescent light from the measurement sample by irradiating laser light on the measurement sample
Data Source
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AI summary
The present invention is to present a cell analyzer capable of measuring cells which are approximately 20 to 100 µm in size with high precision via flow cytometry. The cell analyzer 10 comprises: a flow cell 51 in which a measurement sample including a measurement target cell flows; a light source part 53 for irradiating a light on the measurement sample flowing in the flow cell 51; an optical system 52 for forming a beam spot on the measurement sample flowing in the flow cell 51, the beam spot having a diameter of 3∼8 µm in a flow direction of the measurement sample and a diameter of 300∼600 µm in a direction perpendicular to the flow direction of the measurement sample; and a light receiving part 55, 58, 59 for receiving a light from the measurement sample.