Fluidic flow cytometry devices and methods

• US 9,645,010 B2
• Filed: 08/23/2012
• Issued: 05/09/2017
• Est. Priority Date: 03/10/2009
• Status: Active Grant

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: NanoCellect Biomedical, Inc., Regents of the University of California (University of California)
• Inventors: Lo, Yu-Hwa, Cho, Sung Hwan, Morachis, Jose, Alaynick, Will, Chuang, Kendall, Kim, Nam
• Primary Examiner(s): Chowdhury, Tarifur
• Assistant Examiner(s): Ahmed, Jamil
• Application Number:
  US13/593,488
• Publication Number:
  US 20130083315A1
• Time in Patent Office: 1,720 Days
• Field of Search: 356 39, 356246, 356301, 356335, 356337, 356417, 356 73, 422 73, 435 5, 435 29, 4352872, 2504921
• CPC Class Codes: G01J 3/28 Investigating the spectrum using colour filters G01J3/51
• CPC Class Codes: G01J 3/46 Measurement of colour; Colour measuring devices, e.g. colorimeters measuring colour temperature G01J5/60
• CPC Class Codes: G01J 3/51 using colour filters
• CPC Class Codes: G01N 15/01 specially adapted for biological cells, e.g. blood cells investigating sedimentation of particle suspensions in blood G01N15/05
• CPC Class Codes: G01N 15/1434 Optical arrangements
• CPC Class Codes: G01N 15/1459 the analysis being performed on a sample stream
• CPC Class Codes: G01N 15/149 specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: G01N 2015/144 Imaging characterised by its optical setup
• CPC Class Codes: G01P 5/26 by measuring the direct influence of the streaming fluid on the properties of a detecting optical wave

Abstract

Devices, systems and methods facilitate analyzing, identifying and sorting particles in fluids, including cytometry devices and techniques. The described techniques can be used in a variety of applications such as in chemical or biological testing and diagnostic measurements. One exemplary flow cytometry device includes a channel that is capable of conducting a fluid containing at least one particle and also capable of allowing light be transmitted to and from the channel. The flow cytometry device also includes a lens that is positioned between the channel and a color filter. The lens directs at least a portion of light transmitted from the channel to the color filter. The color filter includes a plurality of zones, where each zone is adapted to allow transmission of only a particular spectral range of light. The flow cytometry device further includes a detector configured to receive the light that is transmitted through the color filter.

Claims

1. A flow cytometry device, comprising:

a channel capable of conducting a fluid containing at least one particle, the channel further capable of allowing light be transmitted to and from the channel;

a lens positioned between the channel and a single color filter and adapted to direct at least a portion of light transmitted from the channel to the single color filter, the single color filter having a continuously graded transmission spectrum from a first wavelength to a second wavelength without an abrupt boundary, the single color filter positioned at a stationary location with respect to the lens, the lens positioned to direct the light emanating from a corresponding range of positions of the at least one particle to impinge on only a portion of the single color filter as the at least one particle moves along the channel so that the light emanating from the particle at a first range of positions of the particle in the channel is directed by the lens to impinge on only a first portion of the single color filter, and as the particle transitions from the first range of positions to a second range of positions within the channel, the light emanating from the particle is directed to transition from falling on only the first portion to fall only on a second portion of the single color filter that allows transmission of light in a different spectral range of the transmission spectrum compared to the first portion of the single color filter; and

a detector configured to receive the light that is transmitted through the single color filter to produce detection signals in digital form; and

a processor implemented in electronic circuitry to receive the detection signals and configured to process the digital data of the detection signals to determine at least one characteristic of interest of the at least one particle, wherein the processor is further configured to filter the detection signals into a plurality of spectral bands within the transmission spectra of the single color filter, wherein a number of filtered spectral bands and spectral bandwidth of each filtered spectral band is changeable by the processor.

2. The flow cytometry device of claim 1, further comprising:

at least one light source positioned to direct light to the mirror;

one or more mirrors positioned to reflect the light from the light source(s) into the channel, wherein the one or more mirrors are substantially transparent to the light transmitted by the channel; and

an optical filter positioned between the lens and the detector and adapted to substantially block the light from the light source and to substantially transmit the light transmitted from the channel into the single color filter.

3. The flow cytometry device of claim 2, wherein at least one of the one or more mirrors is positioned between one of:

the lens and the detector, and

the lens and the channel.

4. The flow cytometry device of claim 1, further comprising an additional lens positioned between the color filter and the detector.

5. The flow cytometry device of claim 1, wherein each portion of the single color filter allows transmission of a spectral range of light that is non-overlapping with spectral ranges allowed to be transmitted by all other portions.

6. The flow cytometry device of claim 1, wherein at least one portion of the single color filter is adapted to allow transmission of a spectral range of light that is overlapping with transmission spectral range of at least one other portion.

7. An imaging device, comprising:

a stage configured to move in one or more dimensions and configured to receive an object for imaging, wherein at least a portion of the object is capable of transmitting light; and

a lens positioned between the stage and an image sensor and adapted to direct the light emitted by the at least a portion of the object to the image sensor,

wherein the image sensor comprises a plurality of light sensing elements arranged as a plurality of sensing element subsets, each sensing element subset being responsive to a particular spectral range of light incident thereupon,

a single continuously graded color filter positioned between the lens and the image sensor, the single continuously graded color filter having a continuously graded transmission spectrum from a first wavelength to a second wavelength without an abrupt boundary, and

wherein the image sensor is aligned with the lens and the object so as to receive, at a first subset of the sensing elements, a first spectral range of light emitted by the object when the stage positions the object at a first position, and to receive a second spectral range of light emitted by the object at a second subset of sensing elements when the stage positions the object at a second position,

the imaging device further comprising a processor implemented in electronic circuitry to receive detection signals produced by the image sensor and configured to process the detection signals to determine at least one characteristic of interest of the object, wherein the processor is further configured to filter the detection signals into a plurality of spectral bands within the transmission spectra of the single continuously graded color filter, wherein a number of filtered spectral bands and spectral bandwidth of each filtered spectral band is changeable by the processor.

8. The imaging device of claim 7, wherein each portion of the color filter is adapted to allow transmission of only a particular spectral range of light.

9. A method for identifying one or more particles by flow cytometry, comprising:

flowing one or more particles through a channel of a flow cytometry device;

causing the one or more particles to emit light;

receiving light emitted by the one or more particles after transmission through a lens that directs the light to a single color filter, wherein the single color filter comprises a plurality of contiguously graded transmission spectrum from a first wavelength to a second wavelength without an abrupt boundary, and wherein the single color filter is positioned at a stationary location with respect to the lens, the lens positioned to direct the light emanating from a corresponding range of positions of each particle to impinge on only a portion of the single color filter as each particle moves along the channel so that the light emanating from the particle at a first range of positions of the particle in the channel is directed by the lens to impinge on only a first portion of the single color filter, and as the particle transitions from the first range of positions to a second range of positions within the channel, the light emanating from the particle is directed to transition from falling on only the first portion to fall only on a second portion of the single color filter that allows transmission of light in a different spectral range of the transmission spectrum compared to the first portion of the single color filter;

producing digital data associated with the received light; and

processing the digital data to identify the one or more particles, wherein processing the digital data comprises selecting a plurality of dynamically selectable spectral bands, and filtering the digital data into the plurality of spectral bands within the transmission spectra of the single color filter, wherein a number of selected spectral bands and a spectral bandwidth of each selected spectral band is changeable within the spectra of the single color filter.

10. The method of claim 9, further comprising determining velocity of the one or more particles flowing through the channel.

11. The method of claim 10, further comprising directing at least one of the one or more particles through a channel branch based on identity and velocity of the at least one of the one or more particles.

12. The method of claim 9, wherein the one or more particles are identified based on at least a color of the received light.

13. The method of claim 9, wherein a first particle of the one or more particles emits light at a different spectral range than a second particle of the one or more particles.

14. The method of claim 9, wherein

the received light emitted by the one or more particles corresponds to a continuous waveform.

15. The method of claim 14, further comprising modifying velocity of the one or more particles flowing through the channel to facilitate parsing of the data into a particular number of spectral bands.

16. The method of claim 15, wherein at least one of the velocity and the number of spectral bands are specified by a user of the flow cytometry device.

17. A method for identifying a particle by an imaging device, comprising:

causing a particle within a sample affixed to a movable stage to emit light when the movable stage is at a first position;

receiving light emitted by the particle at a first section of the imaging device, the first section being responsive to a first spectral range of light incident thereupon;

using a lens to direct the light emitted by the particle onto an image sensor;

using a single color filter positioned between the lens and image sensor, the single continuously graded color filter having a continuously graded transmission spectrum from a first wavelength to a second wavelength without an abrupt boundary;

moving the stage to one or more additional positions;

receiving light emitted from the particle when the stage is positioned at each of the one or more additional positions at corresponding one or more additional sections of the imaging device, wherein each additional section of the imaging device is responsive to a particular spectral range of light incident thereupon;

processing data corresponding to the light received at the first section and at the one or more additional sections of the imaging device to identify the particle, wherein the processing comprises selecting a plurality of dynamically selectable spectral bands, and filtering the data into the plurality of spectral bands within the transmission spectra of the single color filter, wherein a number of selected spectral bands and a spectral bandwidth of each selected spectral band is a changeable within the spectra of the single color filter.