System and method for immersion flow cytometry
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Publication Number
US-10107746-B2
Publication Date
2018-10-23
Expiration Date
Abstract
An immersion cytometry system (200, 250) having a primary focusing optic immersed in a fluid stream (209) containing suspended particles (212). The system includes a light source (202) configured to illuminate a sensing region in the fluid stream that includes a focus of the primary optic. Light scattered and/or fluoresced from suspended particles passing through the sensing region is focused by an external tube lens on an external detector. The primary optic in one embodiment is a ball lens. In some embodiments, one or more filter/beam splitters on the optical axis reflect a portion of the signal light towards corresponding detectors, each filter being configured to reflect a preselected waveband of light.
Core Innovation
The invention provides an immersion flow cytometry system for analyzing a fluid stream having particles suspended therein. A light source directs light into a sensing region in the fluid stream, and a primary focusing optic is positioned at least partially immersed in the fluid stream to receive light scattered by particles passing through the sensing region. The primary focusing optic and a tube lens define an optical axis for focusing the received scattered and/or fluoresced light onto detectors.
The system uses a position-sensitive detector to identify light focused by the tube lens corresponding to particles suspended in the fluid stream that are not in a region of interest within the sensing region. The position-sensitive detector includes a first field stop with a light deflector and a peripheral light photo detector configured to receive light deflected by the light deflector, and a first photo detector configured to receive light directed to the first field stop that is not deflected.
To separate wavelength bands, the system includes a first chromatic filter/beam splitter positioned along the optical axis and configured to direct a portion of light transmitted along the optical axis to the first field stop, along with a first photo detector for received light not deflected by the peripheral light photo detector. The system further includes a second chromatic filter/beam splitter along the optical axis with a different center wavelength, directing a portion of light to a second field stop, and a second photo detector configured to receive light focused on the second field stop. Signals from the photo detectors are transmitted to a processing system.
Claims Coverage
The independent claim covers a flow cytometry system with an immersed primary focusing optic and tube lens along an optical axis, combined with a position-sensitive detector to distinguish particles not in a region of interest, and with chromatic filter/beam splitters that route different wavelength portions to different field stops and photo detectors. The independent claim includes at least eight inventive structural features, and it further requires transmission of generated signals to a processing system.
Immersed primary focusing optic receiving scattered light in a sensing region
A primary focusing optic positioned at least partially immersed in the fluid stream and positioned to receive light scattered by particles passing through the sensing region.
Tube lens defining an optical axis to focus received light
A tube lens positioned to focus light received from the primary focusing optic, wherein the tube lens and the primary focusing optic define an optical axis.
Position-sensitive detector with field stop and deflector for out-of-region detection
A position-sensitive detector configured to identify light focused by the tube lens corresponding to particles suspended in the fluid stream that are not in a region of interest within the sensing region, the position-sensitive detector comprising a first field stop with a light deflector, and a peripheral light photo detector configured to receive light deflected by the light deflector.
Chromatic filter/beam splitter routing portion of optical-axis light to first field stop
A first chromatic filter/beam splitter positioned along the optical axis configured to direct a portion of light transmitted along the optical axis to the first field stop.
First photo detector for non-deflected light at the first field stop
A first photo detector configured to receive light directed to the first field stop that is not deflected by the peripheral light photo detector.
Second chromatic filter/beam splitter with different center wavelength routing to second field stop
A second chromatic filter/beam splitter positioned along the optical axis configured to direct a portion of light from the optical axis to a second field stop, wherein the first chromatic filter/beam splitter has a different center wavelength than the second chromatic filter/beam splitter.
Second photo detector for light focused on the second field stop
A second photo detector configured to receive light focused on the second field stop.
Signals generated by photo detectors transmitted to a processing system
Wherein the first photo detector and the second photo detector are each configured to generate a signal responsive to the received light and to transmit the generated signals to a processing system.
Overall, the claim coverage centers on an immersion flow cytometry optical train with an immersed primary focusing optic and tube lens, a position-sensitive detector for identifying particles outside a region of interest, and chromatic filter/beam-splitter optics that direct wavelength-dependent light to different field stops and photo detectors, with the resulting signals sent to a processing system.
Stated Advantages
Increased stream diameter to reduce clogging.
Immersed optics increase numerical aperture and sensitivity.
Reduced background noise by eliminating intermediate boundaries.
Enables non-contact operation and operation in harsh environments.
Enables lower power excitation.
Enables autonomous oceanographic phytoplankton monitoring.
Enables environmental and industrial monitoring, as well as space and air-quality monitoring.
Documented Applications
Autonomous oceanographic phytoplankton monitoring.
Environmental monitoring.
Industrial monitoring.
Space monitoring.
Air-quality monitoring.
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