Particle Detection in Cell Compositions via Lysis and Autofluorescence Imaging
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Solution Overview
Problem
Current methods for detecting the presence or absence of particles, such as magnetic beads, in a cell composition are time-consuming, labor-intensive, and often inaccurate, making it difficult to accurately enumerate or detect the number of particles bound or associated with cells.
Innovation Solution
The method involves incubating a sample containing cells and non-cell particles under conditions to induce cell lysis, followed by fluorescence imaging analysis of the output composition to determine the presence or absence of particles, utilizing autofluorescence if applicable, and potentially enriching for non-cell particles using magnetic selection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If current manual methods are used to detect particles in cell composition, then detection can be performed, but the process is time-consuming and labor-intensive
Solution Approach 1:
The patent replaces manual mechanical counting methods with automated fluorescence imaging and image analysis systems. The system uses automated microscopy with fluorescence channels to capture images of particles, and computer algorithms automatically analyze and count particles, eliminating the need for manual inspection and significantly increasing detection throughput
Solution Approach 2:
The patent creates optical copies (images) of the particle sample using fluorescence microscopy. Multiple fluorescence channels capture different aspects of particle characteristics, and these image copies are then analyzed by automated software to determine particle presence and properties, enabling rapid non-contact analysis of multiple samples
2Measurement precision
If current manual methods are used to detect particles in cell composition, then detection can be performed, but the accuracy is often insufficient
Solution Approach 1:
The patent utilizes fluorescence emission at different wavelengths (color channels) to differentiate particles from cells and to identify particle properties. By detecting autofluorescence signals at specific wavelengths and comparing ratios between channels, the system achieves accurate and reliable particle detection even in complex biological samples
Solution Approach 2:
The replacement of manual detection with automated fluorescence imaging and algorithm-based analysis eliminates human error and subjectivity, providing consistent, reproducible, and highly accurate particle detection across multiple samples and operators
3Reliability
If cells are present in the sample during particle detection, then the complete sample can be analyzed, but false positives and negatives increase
Solution Approach 1:
The patent employs cell lysis to transition cells from an intact state to a lysed state, fundamentally changing their optical properties. Lysed cells no longer exhibit the characteristic fluorescence and morphological features of intact cells, allowing the imaging system to clearly distinguish particles from cellular debris and avoid false positives while maintaining accurate particle enumeration
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 a more efficient, accurate, and automated way to detect and enumerate particles in a cell composition, reducing the likelihood of false positives and negatives, and allowing for high-throughput analysis.
Implementation Method 1
incubating a sample comprising at least a portion of an input composition or a sample derived from the input composition, wherein the incubating is under conditions to induce lysis of cells
Implementation Method 2
analyzing the output composition or a portion thereof by fluorescence imaging of the autofluorescence of the one or more non-cell particles
Data Source
AI summary
Provided are methods of detecting, assessing or determining the presence or absence of particles, such as bead particles, present in a cell composition. Also provided are articles of manufacture and kits for use in the methods.