FLUIDIC FLOW CYTOMETRY DEVICES AND METHODS

• US 20130083315A1
• Filed: 08/23/2012
• Published: 04/04/2013
• Est. Priority Date: 03/10/2009
• Status: Active Grant

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Yu-Hwa Lo
• Inventors: Lo, Yu-Hwa, Cho, Sung Hwan, Chuang, Kendall, Alaynick, Will, Morachis, Jose, Kim, Nam
• Granted Patent: US 9,645,010 B2
• Time in Patent Office: Days
• US Class Current: 356/73
• 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 color filter and adapted to direct at least a portion of light transmitted from the channel to the color filter, the color filter comprising a plurality of zones, each zone adapted to allow transmission of only a particular spectral range of light; and

a detector configured to receive the light that is transmitted through the color filter.

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 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 zone allows transmission of a spectral range of light that is non-overlapping with spectral ranges allowed to be transmitted by all other zones.

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

7. A flow cytometry device, comprising:

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

a lens positioned between the fluidic channel and a diffractive component and adapted to direct at least a portion of light transmitted from the fluidic channel to the diffractive component;

an aperture positioned between the diffractive component and a detector and adapted to allow a particular spectral range of light received from the diffractive component to reach the detector based upon the position(s) of the at least one particle within the fluidic channel.

8. The flow cytometry device of claim 7, wherein a first spectral range of light is received at the detector when the at least one particle is at a first position within the fluidic channel and a second spectral range of light is received at the detector when the at least one particle is at a second position within the fluidic channel.

9. 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, 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 object is 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 object is at a second position.

10. The imaging device of claim 9, further comprising a color filter positioned between the lens and the image sensor, wherein the color filter comprises a plurality of zones, each zone adapted to allow transmission of only a particular spectral range of light.

11. The imaging device of claim 10, wherein the zones are continuously graded in the color filter.

12. The imaging device of claim 10, wherein

the plurality of zones are arranged as different sections of a filter wheel;

and

the filter wheel is configured to rotate and position each of the plurality of zones, one at a time, in an optical path of light transmitted by the object.

13. A method, comprising:

receiving data corresponding to a particular spectral range of light emitted by a particle in a flow cytometry device;

processing the received data to enhance signal-to-noise ratio (SNR) of the received data;

determining particle velocity from the processed data with enhanced SNR;

determining one or more peak parameters for one or more peaks of the processed data with enhanced SNR;

mapping colors in accordance with placement of the peak parameters and/or ratios of the peak parameters; and

determining color of the light emitted by the particle in accordance with the mapping.

14. The method of claim 13, wherein the one or more peak parameters is selected from a group of parameters consisting of:

a peak intensity, a peak width, and a peak area.

15. 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 color filter, wherein the color filter comprises a plurality of zones and each zone is configured to allow transmission of only a particular spectral range of light incident thereupon, and wherein light emitted by each particle at different positions within the channel is incident upon different zones of the color filter;

producing data associated with the received light; and

processing the data to identify the one or more particles.

16. The method of claim 15, further comprising determining velocity of the one or more particles flowing through the channel.

17. The method of claim 16, 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.

18. The method of claim 15, wherein the one or more particles are identified based on at least a color of the received light.

19. The method of claim 15, 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.

20. The method of claim 15, wherein

the color zones of the color filter are continuously graded;

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

the

processing comprises parsing the data into a plurality of spectral bands, wherein the number of spectral bands is changeable.

21. The method of claim 20, 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.

22. The method of claim 21, wherein at least one of the velocity and the number of spectral bands are specified by a user of the flow cytometry device.

23. 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;

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.

24. The method of claim 23, further comprising using a lens to direct the light emitted from the particle onto the imaging device.

25. The method of claim 24, further comprising using a color filter that includes a plurality of zones and is positioned between the lens and the image sensor to allow only a particular spectral range of the light emitted by the particle to reach the imaging device.

26. The method of claim 25, wherein the zones are continuously graded in the color filter.

27. The method of claim 25, wherein:

the plurality of zones are arranged as different sections of a filter wheel; and

using the color filter comprises rotating the wheel to position a first zone in optical path of the light emitted by the particle and rotating the wheel to position a second zone in optical path of the light emitted by the particle.

28. The method of claim 27, wherein identifying the particle is carried out based on a color of light detected at the imaging device.

29. A method, comprising:

flowing at least one particle through a fluidic channel of a flow cytometry device;

causing the at least one particle to emit light;

using a lens to direct the emitted light to a diffractive component, the diffractive component directing a portion of the emitted light to an aperture;

receiving, at a detector, the light is propagated through the aperture, wherein the received light at the detector consists of a particular spectral range dependent at least in-part on position(s) of the at least one particle within the fluidic channel;

processing data corresponding to the light received at the detector to identify the one or more particles.

30. The method of claim 29, receiving the light comprises receiving a first spectral range of light when the at least one particle is at a first position within the fluidic channel and a second spectral range of light when the at least one particle is at a second position within the fluidic channel.

31. The method of claim 29, further comprising determining a velocity of the at least one particle flowing through the channel.

32. The method of claim 31, further comprising directing the at least one particle through a channel branch based on identity and velocity of the at least one particle.

33. The method of claim 29, wherein the at least one particle is identified based on at least a color of the light received at the detector.