Cell Refractive Index Imaging for Apoptosis Viability Detection
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Solution Overview
Problem
Current methods for determining cellular viability in bioreactors, such as those used in pharmaceutical and medical fields, are limited in their ability to accurately differentiate between living and dead cells, particularly in distinguishing between necrosis and apoptosis, which affects production yield.
Innovation Solution
A method utilizing the refractive index of cells, specifically the real and imaginary parts of the refractive index, is employed to determine cellular states by analyzing images through phase microscopy, reconstructing complex light wave propagation, and applying a propagation operator to estimate refractive indices, enabling differentiation between living, dead, and apoptotic states.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If manual counting methods are used to assess cell viability, then device complexity is reduced, but measurement precision and productivity deteriorate
Solution Approach 1:
The patent employs colorimetric indicators that change color based on cell viability status. Live cells and dead cells exhibit distinct color characteristics that can be automatically detected by imaging systems, enabling precise quantitative assessment without complex manual procedures. The color change provides a direct visual and measurable signal of cell viability.
Solution Approach 2:
The invention replaces manual mechanical counting methods with automated optical imaging and image analysis systems. The automated system captures images of the assay wells and uses software algorithms to identify and count cells based on their color and morphological features, eliminating the need for manual microscopy and counting while significantly improving precision and throughput.
2Productivity
If manual counting methods are used to assess cell viability, then device complexity is reduced, but productivity deteriorates
Solution Approach 1:
The invention replaces manual mechanical counting methods with automated optical imaging and image analysis systems. The automated system captures images of the assay wells and uses software algorithms to identify and count cells based on their color and morphological features, eliminating the need for manual microscopy and counting while significantly improving precision and throughput.
Solution Approach 2:
The automated imaging system enables continuous assessment of multiple assay wells in sequence, allowing high-throughput evaluation of cell viability across numerous samples without interruption. The system can process multiple plates sequentially or in parallel, maintaining continuous productive operation rather than the stop-start nature of manual counting.
3Measurement precision
If automated cell counting devices are used, then measurement precision and productivity are improved, but device complexity and cost increase
Solution Approach 1:
The patent employs colorimetric indicators that change color based on cell viability status. Live cells and dead cells exhibit distinct color characteristics that can be automatically detected by imaging systems, enabling precise quantitative assessment without complex manual procedures. The color change provides a direct visual and measurable signal of cell viability.
Solution Approach 2:
The invention replaces manual mechanical counting methods with automated optical imaging and image analysis systems. The automated system captures images of the assay wells and uses software algorithms to identify and count cells based on their color and morphological features, eliminating the need for manual microscopy and counting while significantly improving precision and throughput.
4Productivity
If traditional assays require multiple separate steps, then measurement precision may be maintained, but productivity and ease of operation deteriorate
Solution Approach 1:
The invention merges cell lysis, indicator release, and viability indication into a single integrated assay step. The lytic indicator is incorporated into the assay medium, so when cells are lysed (either naturally or induced), the indicator is released and immediately provides a colorimetric signal of viability. This eliminates separate lysis and measurement steps while maintaining accurate viability assessment.
Solution Approach 2:
The assay system is designed to be universally applicable to different cell types and viability assessment needs. The same basic assay platform and imaging system can evaluate various cell lines, different viability conditions, and multiple parameters simultaneously, allowing one system to perform multiple functions without requiring separate specialized procedures for each application.
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 provides accurate differentiation between living, dead, and apoptotic cells, enhancing the detection of cellular states and improving production yield by identifying apoptotic cells before death occurs.
Implementation Method 1
the cells are lysed and the lytic indicator is released, thereby providing an indication that the cells are dead
Data Source
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AI summary
A method for determining a state of a cell, the cell being placed in a sample, in contact with a culture medium, the method comprising: illuminating the sample with a light source and acquiring an image of the sample with an image sensor, the image sensor lying in a detection plane; from the acquired image, locating a position of the cell in a plane parallel to the detection plane; the method further comprising: from the acquired image, estimating a refractive index of the cell or a relative refractive index of the cell, the relative refractive index corresponding to a refractive index of the cell relative to the refractive index of the culture medium; from the estimation of the refractive index or of the relative refractive index, determining an index of interest of the cell; from the index of interest, classifying a state of the cell among predetermined states, the predetermined states comprising at least one apoptosis state and one living state.