Dual-Mode Cell Imaging Method for Fluorescence Bleaching Reduction

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Solution Overview

Problem

Existing dispensing devices for monoclonal cell lines are inefficient in rapidly processing large numbers of cells, particularly fluorescent and small/transparent cells, due to long exposure times required for accurate imaging, which leads to bleaching and loss of cells during laboratory operations.

Innovation Solution

A method utilizing two measurement signals, where a quick brightfield measurement determines cell presence and a longer fluorescent measurement is only conducted when necessary, allowing for rapid examination and precise detection of cells and particles, including fluorescent ones, while minimizing exposure time to prevent bleaching.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If long exposure time is used for fluorescence imaging, then measurement precision is improved, but productivity deteriorates due to slow cell processing speed

Engineering Contradiction:
Improvefluorescence detection accuracyVSAvoidcell processing speed
Core Design Contradiction:
Measurement precisionVSProductivity

Solution Approach 1:

The patent segments the imaging process into two distinct modes: brightfield imaging for rapid cell localization and fluorescence imaging for precise fluorescent cell identification. This segmentation allows the system to use the appropriate imaging mode for each specific task, thereby resolving the contradiction between speed and precision.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The system performs preliminary brightfield imaging to quickly locate and identify cell positions before conducting fluorescence imaging. This preliminary action reduces the overall exposure time required for fluorescence imaging by pre-positioning the field of view, thus improving both productivity and maintaining measurement precision.

Inventive Principle:
Principle #10Preliminary action

2Measurement precision

If long exposure time is used for fluorescence imaging, then measurement precision is improved, but reliability deteriorates due to fluorescent dye fading

Engineering Contradiction:
Improvefluorescence signal qualityVSAvoidfluorescent dye stability
Core Design Contradiction:
Measurement precisionVSReliability

Solution Approach 1:

The patent implements periodic action by using brief, intermittent fluorescence imaging only when necessary, rather than continuous long-duration exposure. The system alternates between brightfield mode for navigation and short fluorescence exposures for identification, reducing cumulative exposure time and preventing dye fading while maintaining sufficient signal quality.

Inventive Principle:
Principle #19Periodic action

Solution Approach 2:

The system rushes through the fluorescence imaging process by using the minimum necessary exposure time to capture sufficient signal. By skipping unnecessary prolonged exposure and using only the essential brief fluorescence captures, the system maintains measurement precision while preserving fluorescent dye integrity.

Inventive Principle:
Principle #21Skipping (Rushing through)

3Productivity

If rapid dispensing is implemented, then productivity is improved, but measurement precision deteriorates due to insufficient time for accurate cell identification

Engineering Contradiction:
Improvedispensing speedVSAvoidcell location accuracy
Core Design Contradiction:
ProductivityVSMeasurement precision

Solution Approach 1:

The patent segments the cell identification process into two stages: rapid brightfield-based preliminary identification for speed, followed by selective fluorescence-based confirmation for precision. This segmentation enables the system to maintain high dispensing speed while ensuring accurate cell location and identification through the combination of both imaging modes.

Inventive Principle:
Principle #1Segmentation

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 rapid and accurate dispensing of cells into specific locations based on their properties, reducing cell loss and bleaching, and improving the efficiency of cell processing in laboratory operations.

Implementation Method 1

a first measurement method by means of which a first measurement signal can be generated rapidly, in particular by means of a brightfield method

Methodology Applied
Scientific EffectBrightfield imaging:

Implementation Method 2

a second measurement method by means of which a second measurement signal can be generated, in particular by means of a fluorescence method

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Data Source

PatentEP3649501B1Method for examining a liquid comprising at least one cell and/or at least one particle
Publication Date: 2024.08.21 CYTENA GMBH
  • EP3649501B1 patent drawingFigure 1
  • EP3649501B1 patent drawingFigure 2
  • EP3649501B1 patent drawingFigure 3

AI summary

The invention relates to a method for examining a liquid which contains at least one cell and/or at least one particle using a first and a second measurement signal emanating from a liquid region. The method has the following steps: a. reading the first measurement signal and analyzing the first measurement signal and b. using the analyzed first measurement signal to determine whether the second measurement signal was detected and completely read, whether the detected second measurement signal was completely read, or whether the reading of the detected second measurement signal (20) was interrupted.