Flow Cytometer Laser Assessor for Optical Noise Compensation
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Solution Overview
Problem
Flow cytometers face challenges in characterizing analytes in biological fluids due to optical noise and drift caused by fluctuations in laser intensity, which affect the accuracy of particle characterization and separation.
Innovation Solution
The implementation of a laser assessor configured with a reference detector and a non-scattered light selection element allows for dynamic assessment of laser operational changes, enabling compensation for optical noise and drift in flow cytometer data.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Illumination intensity
If a flow cytometer uses a laser to irradiate particles in a flow stream, then light scattering and fluorescence emission occur for particle characterization, but laser intensity fluctuations cause optical noise and drift that degrade measurement precision
Solution Approach 1:
The patent extracts the harmful laser light that passes through the flow cell without scattering off particles by using a beam stop element to block this direct laser beam. This extracted light is then directed to a separate detector that monitors laser intensity fluctuations, allowing the system to identify and compensate for noise without interfering with the primary particle characterization measurements.
Solution Approach 2:
The patent introduces an intermediary detection system consisting of a beam stop element and a dedicated detector that acts as a mediator between the laser and the flow cell. This intermediary component captures the laser light that would otherwise pass through the flow cell and directs it to a monitoring detector, enabling indirect measurement of laser intensity without affecting the main flow cytometry analysis.
2Measurement precision
If the flow stream is irradiated with laser light, then particle characteristics can be characterized through light scattering, but temporal fluctuations in laser intensity due to temperature, pressure, and humidity changes create optical noise
Solution Approach 1:
The patent implements a feedback mechanism where the detector continuously monitors laser intensity fluctuations caused by environmental changes. The system uses this real-time feedback information to compensate for the optical noise in the flow cytometer data, allowing the measurement precision to be maintained despite temperature, pressure, and humidity variations.
Solution Approach 2:
The system performs self-diagnosis by using the beam stop element and detector to continuously monitor the laser's own intensity fluctuations. This self-monitoring capability allows the flow cytometer to detect and correct for environmental effects on the laser without external intervention, maintaining stable measurements throughout operation.
3Measurement precision
If a beam stop element is used to block non-scattered laser light, then direct laser light is prevented from reaching the forward scatter detector, but this requires additional optical components and alignment
Solution Approach 1:
The patent segments the optical path into two separate channels: one for detecting scattered light from particles (forward scatter) and another for monitoring direct laser light intensity. By using a beam stop element to physically separate these paths, the system can independently optimize each detection function without interference, improving measurement precision while managing complexity through functional separation.
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 solution enhances the accuracy of flow cytometer data by isolating and compensating for fluctuations in laser intensity, thereby improving the resolution and reliability of particle characterization.
Implementation Method 1
a reference detector (e.g., a photomultiplier tube) configured to detect non-scattered laser light (i.e., direct laser light or light received directly from a laser) that is not scattered by particles in the flow cell after passing therethrough and produce a reference signal in response thereto
Implementation Method 2
The flow cytometer also includes a non-scattered light selection element that is configured to selectively allow non-scattered laser light to reach the reference detector while preventing channels of particle-modulated light from reaching the reference detector
Implementation Method 3
Laser light irradiating a particle in a flow cell produces multiple channels of light. Light scattered in the forward direction (i.e., forward scattered light) passes through a lens and pinhole before it is directed to a forward scatter detector
Implementation Method 4
Particles that have been excited by the laser emit fluorescent light
Data Source
AI summary
Aspects of the present disclosure include flow cytometers configured to compensate for optical noise caused by operational change of a laser. Flow cytometers according to certain embodiments include a laser assessor configured to assess operational change of a laser. In embodiments, the laser assessor includes a reference detector and a mirror positioned between the reference detector and the flow cell that is configured to reflect forward scattered light to a forward scatter detector and allow non-scattered laser light to pass through to the reference detector. Methods for assessing laser functionality and, where desired, dynamically adjusting flow cytometer data, based on laser reference data from the laser assessor are also provided.