Method and Apparatus for High Throughput Diagnosis of Diseased Cells With Microchannel Devices

Abstract

The method and apparatus of the present invention detects changes in cell biomechanics caused by any of a variety of diseases and conditions. In one embodiment, the method and apparatus of the invention detect infection of red blood cells. In one embodiment, the invention is a method and apparatus comprising a microfluidic channel with a constriction, for trapping infected red blood cells while allowing healthy red blood cells to deform and pass through the channel. In another embodiment, the invention comprises a suspended microchannel resonator for detecting and counting red blood cells at the constriction of the microfluidic channel.

1 . A method of detecting an abnormality in a cell, comprising:
 introducing the cell into a suspended microchannel resonator, wherein the resonator comprises a constriction.   
   
   
       2 . The method of  claim 1 , comprising:
 detecting the cell within the microchannel.   
   
   
       3 . The method of  claim 1 , wherein the cell is a red blood cell. 
   
   
       4 . The method of  claim 1 , wherein the cell is healthy. 
   
   
       5 . The method of  claim 1 , wherein the cell is infected with an infectious agent. 
   
   
       6 . The method of  claim 1 , wherein the cell is infected with a Plasmodium parasite. 
   
   
       7 . The method of  claim 1 , wherein the cell is infected with  Plasmodium falciparum.    
   
   
       8 . The method of  claim 1 , wherein the cell is exposed to disease. 
   
   
       9 . The method of  claim 1 , comprising:
 capturing the cell within the constriction of the microchannel.   
   
   
       10 . The method of  claim 9 , wherein the cell has a mass, and wherein the mass changes the resonance frequency of the suspended microchannel. 
   
   
       11 . The method of  claim 10 , comprising:
 measuring the mass of the cell.   
   
   
       12 . The method of  claim 10 , comprising:
 flowing cells through the microchannel at a rate of at least 3 cells/second; and   detecting diseased cells within approximately 1 minute, wherein at least 1% of the cells are infected.   
   
   
       13 . The method of  claim 1 , wherein the microchannel is approximately 8 microns wide. 
   
   
       14 . The method of  claim 1 , wherein the microchannel is approximately 3 microns high. 
   
   
       15 . The method of  claim 1 , wherein the microchannel is approximately 3 microns wide at the constriction. 
   
   
       16 . An apparatus for detecting an abnormality in a cell, comprising:
 a suspended microchannel resonator, wherein the microchannel comprises a constriction.   
   
   
       17 . The apparatus of  claim 16 , wherein the microchannel detects the presences of the cell. 
   
   
       18 . The apparatus of  claim 16 , wherein the cell is a red blood cell. 
   
   
       19 . The apparatus of  claim 16 , wherein the cell is healthy. 
   
   
       20 . The apparatus of  claim 16 , wherein the cell is infected with an infectious agent. 
   
   
       21 . The apparatus of  claim 16 , wherein the cell is infected with a malaria parasite. 
   
   
       22 . The apparatus of  claim 16 , wherein the cell is infected with  Plasmodium falciparum.    
   
   
       23 . The apparatus of  claim 16 , wherein the cell is exposed to disease. 
   
   
       24 . The apparatus of  claim 20 , wherein the cell is captured by constriction of the micro channel. 
   
   
       25 . The apparatus of  claim 24 , wherein the cell has a mass, and wherein the mass changes the resonance frequency of the suspended microchannel. 
   
   
       26 . The apparatus of  claim 25 , wherein the change in resonance frequency measures cell mass. 
   
   
       27 . The method of  claim 25 , wherein the change in resonance frequency detects disease. 
   
   
       28 . The apparatus of  claim 16 , wherein the microchannel is approximately 8 microns wide. 
   
   
       29 . The apparatus of  claim 16 , wherein the microchannel is approximately 3 microns high. 
   
   
       30 . The apparatus of  claim 16 , wherein the microchannel is approximately 3 microns wide at the constriction. 
   
   
       31 . The apparatus of  claim 27 , wherein cells flow through the microchannel at a rate of at least 3 cells/second. 
   
   
       32 . The apparatus of  claim 31 , wherein diseased cells are detected within approximately 1 minute, and wherein at least 1% of the cells are infected. 
   
   
       33 . The apparatus of  claim 16 , comprising:
 a suspended microchannel resonator reader for detecting and recording oscillations of the resonating microchannel.   
   
   
       34 . The apparatus of  claim 33 , comprising:
 a solid state laser; and   a photodetector for measuring the resonance frequency of the suspended microchannel.   
   
   
       35 . The apparatus of  claim 33 , comprising frequency counters for measuring the resonance frequency of the suspended microchannel. 
   
   
       36 . The apparatus of  claim 33 , comprising a “self-resonant” circuit for maintaining the resonance frequency of the suspended microchannel at a constant. 
   
   
       37 . The apparatus of  claim 33 , comprising a means for controlling the suspended resonance micro channel. 
   
   
       38 . The apparatus of  claim 33 , comprising a means for acquiring and recording data from the suspending resonance microchannel. 
   
   
       39 . The apparatus of  claim 16 , comprising fluidic interfaces for introducing solutions to the micro channel. 
   
   
       40 . The apparatus of  claim 39 , wherein the fluidic interface comprises a pump. 
   
   
       41 . The apparatus of  claim 39 , wherein the fluidic interface comprises a valve. 
   
   
       42 . The apparatus of  claim 16 , wherein suspended microchannel resonator has a length of approximately 300 mm. 
   
   
       43 . The apparatus of  claim 16 , wherein the suspended microchannel resonator has a resonant frequency of approximately 220 kHz. 
   
   
       44 . The apparatus of  claim 16 , wherein the suspended microchannel resonator has an estimated quality factor of great than 2000. 
   
   
       45 . The apparatus of  claim 16 , wherein the suspended microchannel resonator has an estimated frequency resolution of approximately 0.1 ppm. 
   
   
       46 . The apparatus of  claim 16 , wherein the suspended microchannel resonator has an mass resolution of approximately 35 fg. 
   
   
       47 . The apparatus of  claim 16 , wherein the suspended microchannel resonator has an mass resolution for proteins of approximately 140 fg. 
   
   
       48 . A method of detecting or monitoring a disease or clinical condition in a subject comprising the steps of:
 (a) providing a sample containing cells obtained from the subject;   (b) flowing the cells through at least a portion of a microchannel; and   (c) detecting abnormal trapping of one or more of the cells in the microchannel.   
   
   
       49 . The method of  claim 48 , comprising altering the velocity of the cells flowing through at least a portion of the microchannel. 
   
   
       50 . The method of  claim 49 , comprising receptors within the microchannel, and wherein the receptors alter the velocity of the cells flowing through at least a portion of the microchannel. 
   
   
       51 . The method of  claim 49 , comprising controlling the surface charge of the microchannel to alter the velocity of the cells flowing through at least a portion of the microchannel. 
   
   
       52 . A method of detecting or monitoring a disease or clinical condition in a subject comprising:
 (a) providing a sample containing cells obtained from the subject; and   (b) detecting an abnormality in the stiffness of the cells, wherein the presence or extent of the abnormality provides information regarding the presence or severity of the disease or clinical condition.   
   
   
       53 . The method of  claim 52 , further comprising the step of administering or modifying therapy based on the presence or extent of an abnormality or infection. 
   
   
       54 . An apparatus for detecting or monitoring a disease or clinical condition in a subject comprising:
 a microchannel, wherein a sample containing cells obtained from the subject are flowed through at least a portion of the microchannel.   
   
   
       55 . The apparatus of  claim 54 , comprising a means for measuring mass. 
   
   
       56 . The apparatus of  claim 54 , comprising a means for measuring velocity.