Imaging Flow Cytometry for High-Throughput Cell FISH Analysis
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Solution Overview
Problem
Current methods for cytogenetic analysis, such as FISH and immunophenotyping, are limited by low sensitivity and require manual examination, making them inefficient for analyzing large numbers of cells and identifying specific genetic abnormalities.
Innovation Solution
The immuno-flowFISH technique combines immunophenotyping and FISH analysis using imaging flow cytometry to identify cells based on their antigen expression, allowing for automated high-throughput analysis of genomic aberrations in cells of interest.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If manual fluorescent microscopy is used to examine FISH probe location, then chromosomal abnormalities can be detected, but the number of cells analyzed is limited (less than 500 per case) resulting in low sensitivity
Solution Approach 1:
The patent replaces manual fluorescent microscopy with automated flow cytometry to analyze FISH probe signals. Flow cytometry uses optical detection systems with lasers and photodetectors to automatically measure fluorescence intensity in thousands of cells, eliminating the need for manual microscopy and enabling high-throughput analysis while maintaining detection accuracy
Solution Approach 2:
The patent changes the detection parameter from visual inspection of fluorescent images to quantitative measurement of fluorescence intensity by flow cytometry. This parameter change enables automated analysis of cell suspension samples, allowing thousands of cells to be analyzed rapidly with precise measurement of chromosomal abnormality signals
2Measurement precision
If slide-based FISH is performed, then chromosomal abnormalities can be visualized, but the method is labour intensive and does not specifically identify the cell of interest
Solution Approach 1:
The patent replaces manual slide examination with automated flow cytometry analysis. Flow cytometry instruments automatically detect and quantify FISH probe signals in cell suspensions, eliminating labour-intensive manual microscopy while providing precise visualization of chromosomal abnormalities through fluorescent intensity measurements
Solution Approach 2:
The patent transitions from two-dimensional slide-based visualization to three-dimensional flow cytometric analysis of cell suspensions. This dimensional change allows automated detection of chromosomal abnormalities in cells of interest by analyzing fluorescent signals in suspension, improving both ease of operation and specificity
3Measurement precision
If immunophenotyping is combined with FISH (immunoFISH), then specificity of detection is increased, but the number of cells analyzed remains limited due to manual microscopy requirement
Solution Approach 1:
The patent merges immunophenotyping and FISH analysis into a single flow cytometry assay. By combining fluorescently labeled antibodies for immunophenotyping with FISH probes for chromosomal analysis, the method simultaneously detects cell phenotype and genotype in the same cell population, achieving high specificity and high throughput
Solution Approach 2:
The patent creates a multi-functional flow cytometry system that performs both immunophenotyping and FISH analysis in a single instrument and protocol. This universal approach allows simultaneous detection of multiple parameters including cell surface markers and chromosomal abnormalities, maximizing both specificity and productivity
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 higher sensitivity and accuracy in detecting chromosomal abnormalities by analyzing thousands of cells, enabling early detection of diseases and informing treatment decisions.
Implementation Method 1
analysing the cell population's antigenic profile by immunophenotyping the population to detect the presence or absence of one or more biological markers
Implementation Method 2
FISH is based on fluorescently-labelled single-stranded DNA probe annealing to its complementary sequence in a target genome to detect DNA sequences and sub-microscopic genetic changes
Implementation Method 3
immunophenotyping by conventional flow cytometry is an automated cell phenotyping method that analyses thousands of cells
Implementation Method 4
Interphase FISH-IS-labelled whole cells can be analysed by flow cytometry enabling automated genetic analysis of several log more cells than microscopy-based approaches
Data Source
AI summary
The present invention provides a method for cell analysis, comprising: preparing a blood sample comprising nucleated cells having surface, cytoplasmic or nuclear antigens (markers); antibody staining the cell markers; fixing and permeabilising the cells; FISH probe hybridising to chromosomes in the cells; performing imaging flow cytometry on the cells; analysing data obtained from performing imaging flow cytometry; and diagnosing, prognosing or monitoring a medical condition based on the data analysis.