Mitochondrial Morphology Assessment via Flow Cytometry
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Solution Overview
Problem
Current methods lack a simple and efficient way to assess the morphological status of mitochondria in isolated cells, which is crucial for molecular biology assays but often requires time-consuming high-resolution microscopy and is not representative of the entire sample.
Innovation Solution
A method involving fluorescent labeling of mitochondria, measuring light from cells in a flow stream, generating images, calculating radial moment image parameters, and assessing morphology based on these parameters to quickly evaluate mitochondrial morphology and viability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If high resolution microscopy is used to visualize cell mitochondria, then measurement precision of mitochondrial morphology is improved, but device complexity and time consumption increase
Solution Approach 1:
The patent replaces complex mechanical microscopy systems with a flow cytometry-based optical detection system. By using fluorescent dyes to label mitochondria and detecting fluorescence signals as cells flow through a detection region, the system achieves mitochondrial morphology assessment without requiring complex mechanical focusing and imaging mechanisms, thus reducing device complexity while maintaining measurement capability
Solution Approach 2:
The patent transforms the assessment parameter from direct visual imaging to quantitative fluorescence signal parameters. By measuring fluorescence intensity, distribution, and temporal characteristics as cells pass through the detection region, the system converts morphological information into measurable physical parameters that can be processed electronically, simplifying the overall system
2Measurement precision
If high resolution microscopy is used to visualize cell mitochondria, then measurement precision of mitochondrial morphology is improved, but loss of time increases
Solution Approach 1:
The patent implements continuous flow-through detection where cells are continuously passed through the detection region in a fluid stream. The fluorescence detection operates continuously as cells pass by, allowing multiple cells to be assessed in sequence without interruption, thereby dramatically reducing the total assessment time compared to sequential manual microscopy examination
Solution Approach 2:
The patent performs fluorescent labeling of mitochondria in advance before the flow-through assessment. By pre-labeling the cells with mitochondrial-specific fluorescent dyes, the actual measurement process only requires detecting the pre-established fluorescence signals, eliminating the need for time-consuming sample preparation during the assessment phase
3Device complexity
If limited number of images are taken for mitochondrial visualization, then device complexity is reduced, but measurement precision and representativeness worsen
Solution Approach 1:
The patent leverages the natural flow characteristics of the cell suspension to automatically present multiple cells in sequence through the detection region. The fluid stream itself serves to deliver a continuous series of cells, eliminating the need for manual image acquisition and selection, and ensuring that the assessment is based on a statistically representative sample of the entire cell population
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 allows for rapid and precise assessment of mitochondrial morphology, increasing the precision of determining cell viability and dynamics by up to 99% and preparing suitable cells for downstream assays within minutes.
Implementation Method 1
measuring light from cells in a sample having fluorescently-labeled mitochondria
Implementation Method 2
Droplets are passed through an electrostatic field and are deflected based on polarity and magnitude of charge on the droplet into one or more collection containers
Data Source
AI summary
Aspects of the present disclosure include methods for assessing morphology of cell mitochondria (e.g., for use to determine viability of cells of a sample). Methods according to certain embodiments include measuring light from cells in a sample having fluorescently-labeled mitochondria, generating images of the cell mitochondria from the measured light, calculating an image parameter from the generated images of the cell mitochondria and assessing morphology of the cell mitochondria based on the calculated image parameter. Systems and integrated circuit devices (e.g., a field programmable gate array) for practicing the subject methods are also described. Non-transitory computer readable storage media are also provided.