Cartridge for Bead-Milling-Based Testing of Erythrocyte Mechanical Fragility

Abstract

A bead mill and an associated bead-mill-based machine for testing mechanical fragility of red blood cells, employing a cartridge configured to contain a sample while cells get stressed via bead oscillation and, in the case of the fragility testing machine, also while lysis levels get detected, for presentation of fragility information.

We claim: 
     
         1 . A cartridge for use in testing mechanical fragility of red blood cells (RBC), the cartridge comprising:
 a body having
 a first portion configured for securely mounting the cartridge within a mechanical fragility testing device; 
 a second portion defining a chamber for holding a sample containing red blood cells (RBC); 
 first and second end portions disposed at opposite longitudinal ends of the second portion of the body and enclosing each end of the chamber, at least one of the first and second end portions defining a sealable passage for inserting the sample into the chamber; 
   a bead disposed within the chamber and configured for movement that progressively lyses the RBC within the chamber in response to forces imparted by the mechanical fragility testing device;   wherein the second portion of the body includes a flexible, transparent portion configured to be in contact with at least a representative portion of the sample, the flexible, transparent portion configured to be pinched and permit repeated projections of light therethrough, while the sample and the bead are within the chamber, to produce optical measurements for ascertaining levels of hemolysis present in the sample at a plurality of points in time.   
     
     
         2 . The cartridge of  claim 1 , wherein the end portions comprise plugs. 
     
     
         3 . The cartridge of  claim 1 , wherein said flexible, transparent portion comprises a flexible plastic tube for containing the sample. 
     
     
         4 . The cartridge of  claim 3 , further comprising a rigid tube that substantially encases the flexible plastic tube. 
     
     
         5 . The cartridge of  claim 1 , wherein the chamber is substantially without air when sealed. 
     
     
         6 . The cartridge of  claim 1 , wherein said movement is oscillation at a frequency of up to 30 Hz 
     
     
         7 . The cartridge of  claim 1 , wherein said forces are magnetic. 
     
     
         8 . The cartridge of  claim 7 , wherein said forces are from an audio transducer in the testing device. 
     
     
         9 . The cartridge of  claim 1 , wherein the bead has grooves, ridges, a textured surface, and/or varying diameters along its length. 
     
     
         10 . The cartridge of  claim 1 , wherein the bead is coated for biocompatibility with the sample. 
     
     
         11 . A cartridge for use in testing mechanical fragility of red blood cells (RBC), the cartridge comprising:
 a body having
 a first portion configured for securely mounting the cartridge within a mechanical fragility testing device; 
 a second portion defining a plurality of chambers for holding respective subsamples, the subsamples each comprising a portion of a sample containing red blood cells (RBC); 
 first and second end portions disposed at opposite longitudinal ends of the second portion of the body and enclosing each end of each chamber of the plurality of chambers, at least one of the first and second end portions defining a sealable passage for inserting the sample into the second portion of the body and splitting the sample into the subsamples in each chamber of the plurality of chambers; 
   a plurality of beads, each bead disposed within a respective chamber of the plurality of chambers and configured for movement that progressively lyses the RBC within the respective chamber in response to forces imparted by the mechanical fragility testing device;   wherein the second portion of the body includes for each chamber a flexible, transparent portion configured to be in contact with at least a representative portion of the subsample contained in the chamber, the flexible, transparent portion configured to be pinched and permit repeated projections of light therethrough, while the subsample and a corresponding bead of the plurality of beads are within the chamber, to produce optical measurements for ascertaining levels of hemolysis present in the subsample at a plurality of points in time.   
     
     
         12 . The cartridge of  claim 11 , wherein the end portions comprise plugs. 
     
     
         13 . The cartridge of  claim 11 , wherein the flexible, transparent portion of each chamber comprises a flexible plastic tube for containing the subsample in the chamber. 
     
     
         14 . The cartridge of  claim 13 , further comprising a plurality of rigid tubes, each of the plurality of rigid tubes substantially encasing a corresponding flexible plastic tube. 
     
     
         15 . The cartridge of  claim 11 , wherein each chamber is substantially without air when said second portion of the body is sealed. 
     
     
         16 . The cartridge of  claim 11 , wherein said movement is oscillation at a frequency of up to 30 Hz. 
     
     
         17 . The cartridge of  claim 11 , wherein said forces are magnetic. 
     
     
         18 . The cartridge of  claim 17 , wherein said forces are from an audio transducer in the testing device. 
     
     
         19 . The cartridge of  claim 11 , wherein at least one bead of the plurality of beads has grooves, ridges, a textured surface, and/or varying diameters along its length. 
     
     
         20 . The cartridge of  claim 11 , wherein at least one bead of the plurality of beads is coated for biocompatibility with the sample.