Image-based cell sorting systems and methods

Abstract

Disclosed are systems, devices and methods for imaging and image-based sorting of particles in a flow system, including cells in a flow cytometer. In some aspects, a system includes a particle flow device to flow particles through a channel, an imaging system to obtain image data of a particle during flow through the channel, a processing unit to determine a property associated with the particle and to produce a control command for sorting the particle based on sorting criteria associated with particle properties, and an actuator to direct the particle into one of a plurality of output paths of the particle flow device in real-time.

1 .- 20 . (canceled) 
     
     
         21 . An image-based flow cytometry system, comprising:
 a particle flow device structured to include a substrate comprising a channel, the particle flow device operable to flow cells along a flow direction to a first region of the channel;   an imaging system configured to obtain optical signal data associated with a cell when the cell is in the first region; and   a data processing and control unit comprising a processor, the data processing and control unit in communication with the imaging system and configured to (i) process the optical signal data into a two-dimensional image of the cell, (ii) extract one or more parameters from the two-dimensional image of the cell, wherein the one or more parameters are associated with one or more spatial features of the cell,   wherein the imaging system is configured to obtain the optical signal data at a sampling rate of no less than 200 kHz and the cell is flowed through the channel at a speed of no less than 0.08 m/s.   
     
     
         22 . The system of  claim 21 , wherein the imaging system includes one or more light sources to provide an input light to the cell at the first region of the particle flow device, and an optical imager configured to receive optical image data from the cells in the first region. 
     
     
         23 . The system of  claim 22 , wherein the optical imager comprises a spatial filter, the spatial filter comprising a plurality of slits, and wherein a resolution of the two-dimensional image of the cell is diffraction limited in the flow direction with respect to a length of one or more slits of the plurality of slits in the flow direction. 
     
     
         24 . The system of  claim 21 , wherein the optical signal data comprises a first optical signal and a second optical signal, the first optical signal comprising a first fluorescent signal, and the second optical signal comprising a second fluorescent signal. 
     
     
         25 . The system of  claim 21 , wherein the optical signal data comprises a first optical signal and a second optical signal, the first optical signal comprising a first fluorescent signal, and the second optical signal comprises a visible light signal. 
     
     
         26 . The system of  claim 21 , wherein the data processing and control unit is configured to process the image data to produce the image data set by filtering the image data, reconstructing a first image based on the filtered data, and resizing the reconstructed first image to produce a second image, wherein the second image includes binary image data. 
     
     
         27 . The system of  claim 21 , wherein the data processing and control unit is configured to determine, from the two-dimensional image of the cell, one or more of an amount or a size of a feature of or on the cell, one or more particles attached to the cell, or a particular morphology of the cell or portion of the cell. 
     
     
         28 . The system of  claim 21 , wherein the one or more parameters from the two-dimensional image of the cell comprise at least one of fluorescence area, bright field area, fluorescence perimeter, or bright field perimeter. 
     
     
         29 . The system of  claim 21 , wherein the data processing and control unit is operable to analyze the cell including analysis criteria, and wherein the analysis criteria include a cell contour, a cell size, a cell shape, a nucleus size, a nucleus shape, a fluorescent pattern, or a fluorescent color distribution. 
     
     
         30 . The system of  claim 29 , wherein the analysis comprises counting the absolute number of cells exhibiting specific analysis criteria. 
     
     
         31 . The system of  claim 29 , wherein the analysis comprises counting the relative number of cells exhibiting specific analysis criteria as compared to the number of cells analyzed. 
     
     
         32 . The system of  claim 21 , wherein the one or more parameters from the two-dimensional image of the cell are associated with a physiological property of the cell including a cell life cycle phase, an expression or localization of a protein by the cell, an expression or localization of a gene by the cell, a damage to the cell, or an engulfment of a substance or a particle by the cell. 
     
     
         33 . The system of  claim 32 , wherein the damage to the cell includes DNA damage. 
     
     
         34 . A method for imaging a particle, comprising:
 directing a light beam to a fluidic channel, such that at least some light of the light beam is scattered by a particle within the fluidic channel or such that at least some of the light of the light beam causes fluorescent emission from the particle in the fluidic channel;   encoding an optical signal using an optical filter, the optical filter defining a plurality of optical paths along which the scattered light or emitted fluorescent light is received from different portions of a particle flowing past the optical paths while flowing through the fluidic channel, wherein the different portions pass different optical paths at different times;   detecting the encoded optical signal using an optical detector; and   processing the detected optical signal at a data processing unit in communication with the optical detector to detect information of one or more physical characteristics of the particle.   
     
     
         35 . The method of  claim 34 , further comprising collecting data regarding the particle based on the one or more physical characteristics of the particle. 
     
     
         36 . The method of  claim 34 , further comprising producing image data based on the detected optical signal. 
     
     
         37 . The method of  claim 36 , further comprising forming an image of the particle based on the image data. 
     
     
         38 . The method of  claim 34 , wherein the detected optical signal is processed in real-time while the particle flows in the fluidic channel. 
     
     
         39 . The method of  claim 34 , wherein the optical detector comprises a photomultiplier tube. 
     
     
         40 . The method of  claim 34 , wherein the optical detector is configured to obtain the optical signal data at a sampling rate of no less than 200 kHz and the particle is flowed through the channel at a speed of no less than 0.08 m/s.