Disposable chip-type flow cell and cell sorter using the same

Abstract

The object of the present invention is to provide (1) a cell sorter, (2) a flow cytometer capable of detecting sideward scattered light, (3) a method for accurately measuring cell concentration, (4) a method for multicolor staining analysis without a fluorescence correction, and the like, which satisfy requirements that carry-over and cross contamination of samples do not occur. The object can be solved by an apparatus for separating particles comprising: a flow cell wherein a flow path is formed in a flat substrate, an illumination unit configured to illuminate the particles in a sample liquid flowing through the flow path, a detection unit configured to detect particles of interest by detecting scattered light or fluorescence from the particle when the particle is illuminated, and identifying the particle based on its signal intensity, a constant-pressure pump which applies a pressure pulse to the particles in the sample liquid flowing through the flow path in the flow cell, and an electromagnetic valve connected thereto, and a control unit configured to control the movement of the electromagnetic valve based on the signal from the detection unit.

1 . A flow cytometer for multiple samples, wherein a sample liquid is illuminated with light while the sample liquid containing biological particles flows through a flow path in the flow cell, and light generated from particles contained in the sample liquid is detected;
 characterized in that:   the flow cytometer comprises:   a stage for placing the flow cell,   an illumination unit,   a detection unit for detecting the biological particles, and   a control unit configured to control movements of the above parts;   the flow cell is flat-plate, wherein multiple sample liquid reservoirs, multiple sheath liquid reservoirs, multiple discharged liquid reservoirs, multiple collected sample liquid reservoirs, and multiple flow paths fluidly connected thereto, are formed on a flat-plate substrate;   each of the sample liquid reservoirs is separately formed in each of sheath liquid reservoirs so as not to mix the liquids,   each of flow paths for sample liquid is connected to each of the sample liquid reservoirs,   a pair of flow paths for sheath liquid are connected to each of the sheath liquid reservoirs,   the pair of flow paths for sheath liquid are connected to a side surface of each of flow paths for sample liquid,   a joining flow path in which sheath flows are joined to a sample flow from the left and right sides of the sample liquid flows is formed, by connecting the pair of flow paths for sheath liquid to the flow path for sample liquid, the joining flow paths are formed so as to be parallel at equally spaced intervals, and the downstream end of the joining flow path is connected to the discharged liquid reservoir and the collected sample liquid reservoir formed on the flow cells; and   each of the flow paths is illuminated in sequence by moving an illumination light relative to the flow cell, or the flow cell relative to the light (by step and repeat) using a light beam that illuminates only one flow path at a time, so that multiple samples are analyzed.   
     
     
         2 . A flow cytometer comprising an illumination light source, and a fluorescence measuring device configured to measure multiple fluorescences at different wavelengths;
 wherein the flow cytometer comprises:   a unit for analyzing and showing a distribution cells, based on the intensity ratio of fluorescence at two different wavelengths generated by illumination with a light, in the analysis of cells having more than one fluorescence, and   a unit for estimating a quantitative ratio of multiple fluorescences in each of the cells based on the intensity ratio.   
     
     
         3 . An apparatus for measuring a biological particle,
 characterized in that the apparatus comprises:   an illumination light source,   and wherein:
 (i) the apparatus additionally comprises:
 a light measuring device configured to measure intensities of light generated from each particle by illumination with light in multiple wavelength ranges, and 
 a unit for analyzing a property of the particle wherein the property of the particle is analyzed by an index obtained by calculating multiple signal intensities corresponding to multiple measurement aspects of the particle; or 
 
 (ii) the apparatus additionally comprises:
 a light measuring device configured to measure intensities of light generated from each cells by illumination with light in multiple wavelength ranges, 
 wherein an index of each particle identified by a mathematical formula using multiple signal intensities is calculated, and the index can be displayed on a graph to illustrate data relating to the particles. 
 
   
     
     
         4 . The apparatus for measuring a biological particle according to  claim 3 ,
 characterized in that the apparatus comprises:   an illumination light source,   a light measuring device configured to measure intensities of light generated from each particle by illumination with light in multiple wavelength ranges, and   a unit for analyzing a property of the particle wherein the property of the particle is analyzed by an index obtained by calculating multiple signal intensities corresponding to multiple measurement aspects of the particle.   
     
     
         5 . The apparatus for measuring a biological particle according to  claim 4 ,
 wherein the apparatus further comprises   a calculation unit for judging whether the biological particle is a dead cell or live cell, based on the value of an index obtained for each particle by calculating an intensity ratio of fluorescence signals at two different wavelengths generated by illumination with a light, when cells or bacteria which are stained simultaneously by a dye capable of penetrating cell membrane and a dye not capable of penetrating cell membrane are judged whether the cells or bacteria are dead or alive.   
     
     
         6 . The apparatus for measuring a biological particle according to  claim 3 ,
 characterized in that the apparatus comprises:   an illumination light source, and   a light measuring device configured to measure intensities of light generated from each cells by illumination with light in multiple wavelength ranges,   wherein an index of each particle identified by a mathematical formula using multiple signal intensities is calculated, and the index can be displayed on a graph to illustrate data relating to the particles.