Cell Sorting via Transmitted Light Analysis
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Solution Overview
Problem
Conventional cell analyzers, such as flow cytometers, face challenges in accurately distinguishing and sorting cells without causing damage, particularly in analyzing cell cycles and identifying specific cells like stem cells, due to the need for staining which can harm cells and reduce analysis accuracy.
Innovation Solution
A method and device using transmitted light information and side-scattering light information to distinguish and sort cells without nuclear staining, allowing for the recognition and separation of living cells at specific phases of the cell cycle, including G1 and M phases, and identifying polyploid-nucleate or alloploid-nucleate cells, without fluorescent labeling or antibody reactions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If nuclear staining is performed to analyze cell cycle phases, then cell cycle analysis accuracy is improved, but cell damage increases and cells may perish
Solution Approach 1:
The invention extracts the harmful staining step from the cell cycle analysis process. By using transmitted light information to directly measure cell size and nuclear characteristics without any staining, the method eliminates the harmful chemical interactions while preserving the ability to distinguish cell cycle phases through physical optical properties alone
Solution Approach 2:
The invention replaces the chemical staining mechanism with a physical optical measurement mechanism. Instead of using chemical dyes that interact with cellular components, the system uses transmitted light intensity and scattering properties to infer cell cycle phase, substituting chemical analysis with physical measurement
2Measurement precision
If fluorescent labeling is used to identify specific cells, then cell identification accuracy is improved, but cell damage increases
Solution Approach 1:
The invention removes the fluorescent labeling step entirely from the cell identification process. By relying on transmitted light information that reflects inherent cellular optical properties such as size, shape, and internal structure, the method identifies specific cell types without introducing external fluorescent chemicals that could harm the cells
Solution Approach 2:
The invention enables cells to serve themselves in the identification process by measuring their intrinsic optical properties. The transmitted light information naturally reveals cellular characteristics without requiring external labels, allowing cells to be identified through their own physical properties rather than through imposed chemical markers
3Productivity
If conventional flow cytometry is used for cell sorting, then sorting capability is achieved, but analysis accuracy for specific cell phases is insufficient
Solution Approach 1:
The invention changes the measurement parameters from conventional scattering light intensity to transmitted light intensity and its distribution characteristics. By analyzing how light transmits through cells at different phases of the cell cycle, the system achieves both accurate identification and effective sorting, resolving the contradiction between sorting capability and analysis accuracy
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 accurate sorting of living cells without damaging them, allowing for real-time study and extraction of stem or cancer stem cells without using fluorescent chemicals, thereby reducing cell damage and improving analysis accuracy.
Implementation Method 1
If the cells that are flowing in line are irradiated with laser light, scattering light or fluorescence are emitted from the cells that are passing
Implementation Method 2
scattering light or fluorescence are emitted from the cells that are passing
Implementation Method 3
A fluorescent pigment absorbs a certain wavelength and converts high energy light (short wavelength) to lower energy light (long wavelength)
Implementation Method 4
Each fluorescent pigment has the unique excitation wavelength distribution and the emission wavelength distribution
Data Source
AI summary
A method for distinguishing and sorting cells characterized by comprising distinguishing and sorting a specific cell mass or a part of the cells in the cell mass with the use of transmitted light data reflecting the morphological characteristics of the cells such as size and shape optionally together with side-scattering light data reflecting the characteristics of the internal structure of the cells. The part of the cells in the specific cell mass as described above are at the G1 stage or at a part of the M stage in the cell cycle. A part of the cells at the G1 stage are referred to as the left bottom line in an analytical dispersion diagram of the cells wherein the abscissa indicates the transmitted light data, while a part of the cells at the M stage are referred to as the right bottom line in the analytical dispersion diagram of the cells wherein the abscissa indicates the transmitted light data.