Flow Cytometer Light Collection Enhancers for Side-Scatter Capture
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Solution Overview
Problem
Conventional flow cytometers fail to detect a substantial portion of side-scattered and fluorescent light emitted by particles in a flow stream due to the light being collected within a single light collection cone, resulting in incomplete data collection.
Innovation Solution
The integration of a light collection enhancer in flow cytometers, comprising a reflective optical element and a condenser lens, redirects and collimates particle-modulated light propagating outside the primary collection cone, allowing it to be back-propagated and focused by the objective lens for detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Device complexity
If a single light collection cone is used in conventional flow cytometers, then the device complexity is reduced, but the amount of detected light is insufficient
Solution Approach 1:
The light collection system is segmented into multiple light collection cones (first light collection cone and second light collection cone) that collect light from different directions. The objective lens is divided into first and second portions, with each portion dedicated to a specific light collection cone, enabling simultaneous collection of light from multiple angular regions around the flow stream.
Solution Approach 2:
The system transitions from collecting light in a single angular dimension to collecting light in multiple angular dimensions by adding a second light collection cone at a different angle relative to the flow stream. This multi-dimensional light collection approach captures side-scattered and fluorescent light that would otherwise propagate outside the single collection cone.
2Ease of operation
If light is emitted in all directions from particles, then the light distribution is complete, but the light collection efficiency is reduced
Solution Approach 1:
The reflective optical element acts as an intermediary to redirect light that would otherwise be lost. It captures light from the second light collection cone and redirects it to the second portion of the objective lens, converting what would be wasted light into useful detected signal without interfering with the primary light collection path.
3Measurement precision
If multiple light collection cones are implemented, then the signal-to-noise ratio is improved, but the device complexity increases
Solution Approach 1:
Light from both the first and second light collection cones is merged and focused by their respective portions of the objective lens onto the same detection plane. This combining of light paths from multiple angular regions increases the total detected signal and improves signal-to-noise ratio while sharing common detection resources.
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 enhancement increases the total amount of detected light, improving the signal-to-noise ratio and data quality by collecting light that would otherwise be undetected, thereby enhancing the sensitivity and accuracy of flow cytometric analysis.
Implementation Method 1
the reflective optical element redirects and collimates particle-modulated light propagating outside the primary collection cone
Implementation Method 2
the condenser lens redirects and collimates particle-modulated light propagating outside the primary collection cone, allowing it to be back-propagated and focused by the objective lens
Implementation Method 3
an objective lens collects the scattered and fluorescent light within a collection cone and focuses the collected light so that it can be detected
Data Source
AI summary
Flow cytometers including light collection enhancers are provided. In embodiments, the subject flow cytometers include a flow cell, a light source, an objective lens for focusing particle-modulated light propagating within a first light collection cone and a light collection enhancer configured to collect particle-modulated light propagating along an optical path within a second light collection cone and redirect the collected light such that it is back-propagated along the same optical path and focused by the objective lens for detection. Light collection enhancers of interest include a reflective optical element (e.g., a mirror) and a condenser lens positioned between the reflective optical element and the flow cell. Methods for analyzing a sample are also provided.