Method and device for collecting and preserving cells for analysis

Abstract

The claimed subject matter comprises a device to collect and preserve cells comprising of: (1) a collection container comprised of a tube having an open end and a closed end, a closure in the open end of the tube, a vacuum drawn to a predetermined level inside the container; and (2) compounds including an anticoagulant agent and a fixative agent, wherein the compounds are in a sufficient amount to preserve said cells” original morphology and antigenic sites without significant dilution of said cells, and thereby allowing said cells to be directly analyzed by a flow cytometer without further treatment. The claimed subject matter further comprises of a method of making a collection device for cells comprising of: (1) providing a tube having an open end and a closed end; (2) preloading compounds including: an anticoagulant agent, and a fixative agent into the tube, wherein the compounds are in a sufficient amount to preserve the cells” original morphology and antigenic sites without significant dilution of the cells, and thereby allowing the cells to be directly analyzed by a flow cytometer without further treatment; (3) inserting a closure into the open end of the tube; and (4) drawing a vacuum inside the tube to a predetermined level to form the collection device.

1 . A method of making a collection device for cells comprising of:
 providing a tube having an open end and a closed end;   preloading compounds including: acid citrate dextrose, and a fixative agent into said tube, said fixative agent including diazolidinyl urea; wherein said compounds are in a sufficient amount to preserve said cells' original morphology and antigenic sites without significant dilution of said cells, and thereby allowing said cells to be directly analyzed by a flow cytometer without further treatment;   inserting a closure into said open end of said tube; and   drawing a vacuum inside said tube to a predetermined level to form said collection device.   
     
     
         2 - 27 . (canceled) 
     
     
         28 . A method for collecting mammalian blood cells, comprising steps of:
 (a) providing a tube including preloaded compounds consisting of
 (i) acid citrate dextrose and 
 (ii) diazolidinyl urea, the tube having an open end and a closed end that receives cells collected directly from a blood draw and wherein a majority of an interior portion of the tube is substantially free of contact with the preloaded components; 
   (b) drawing a blood sample containing a plurality of blood cells into the tube whereby it contacts the preloaded compounds to yield a final composition, wherein a ratio of a volume of the preloaded compounds to a combined volume of the blood sample and the preloaded compounds is from about 1:100 to about 2:100, and so that the plurality of blood cells of the blood sample are stabilized directly and immediately upon the blood draw; and   (c) transporting the blood sample, wherein the blood sample is drawn and transported in the same tube with no processing steps between the blood draw and transporting.   
     
     
         29 . The method of  claim 28 , wherein a concentration of said acid citrate dextrose preloaded in said tube is less than 0.3 g of acid citrate dextrose per ml of the preloaded compounds. 
     
     
         30 . The method of  claim 28 , further comprising a step of sterilizing said preloaded compounds prior to the preloading step. 
     
     
         31 . The method of  claim 28 , further comprising a step of providing at least one component selected from the group consisting of an alcohol swab, a gauze, a tube holder, a tourniquet, a glove, other cell collection tube, a needle, a lancet, an adhesive strip, a syringe, a test strip, a strip containing reagents for cell analysis, a packaging means for storing said at least one component and said tube to form a kit, and a packaging means for transporting said tube immediately upon the blood draw. 
     
     
         32 . A method for preparing blood cells for analysis, said method comprising steps of:
 (a) providing a closed collection container, said collection container having an internal pressure less than atmospheric pressure outside said container, wherein said collection container contains preloaded compounds consisting of:
 (i) acid citrate dextrose; and 
 (ii) diazolidinyl urea; and 
   wherein a majority of an interior portion of the collection container is substantially free of contact with the preloaded components; and   (b) drawing a blood sample containing the blood cells into the collection container whereby the blood sample contacts the preloaded compounds to yield a final composition, wherein after collection of the blood cells in the container, a ratio of a volume of the preloaded compounds to a volume of the final composition is from about 1:100 to about 2:100.   
     
     
         33 . The method of  claim 32 , wherein a concentration of said diazolidinyl urea preloaded in said collection container is less than about 0.5 g of diazolidinyl urea per ml of the preloaded compounds. 
     
     
         34 . A method for preparing blood cells for analysis, said method comprising:
 (a) providing a collection container for receiving a whole blood sample;   (b) introducing into the collection container preloaded compounds consisting of:
 (i) acid citrate dextrose; and 
 (ii) diazolidinyl urea; 
   (c) evacuating the collection container to an internal pressure that is less than atmospheric pressure outside said collection container;   (d) drawing a volume of the whole blood sample into the collection container, wherein a majority of an interior portion of the collection container is substantially free of contact with the preloaded compounds and whereby the whole blood sample contacts the preloaded compounds to yield a final composition, wherein a ratio of a volume of the preloaded compounds to a volume of the final composition is from about 1:100 to about 2:100.   
     
     
         35 . The method of  claim 34 , said method further comprising storing the whole blood sample at ambient temperature in the collection container after contacting with the preloaded compounds and prior to any analyzing step. 
     
     
         36 . The method of  claim 35 , wherein said whole blood sample is stored at ambient temperature for a period of at least 3 days prior to any analyzing step. 
     
     
         37 . The method of  claim 34 , said method further comprising transporting the whole blood sample at ambient temperature in the collection container from a collection site to an analysis site. 
     
     
         38 . The method of  claim 30 , wherein the sterilizing step includes sterile filtration. 
     
     
         39 . The method of  claim 28 , wherein a concentration of the diazolidinyl urea is less than about 0.5 g/ml before the blood draw. 
     
     
         40 . The method of  claim 28 , wherein the transporting step occurs in ambient temperature. 
     
     
         41 . The method of  claim 28 , further comprising analyzing the blood sample of step (c) by a flow cytometer without further dilution and/or treatment. 
     
     
         42 . The method of  claim 28 , further comprising screening the blood sample of step (c) for HIV. 
     
     
         43 . The method of  claim 28 , wherein the tube includes a closure, and the closure and the tube together form a seal capable of maintaining a pressure differential between atmospheric pressure and a pressure less than atmospheric pressure within the tube. 
     
     
         44 . The method of  claim 32 , wherein the tube has an interior surface that is sterile and resists adherence to the blood cells during collection, storage, and analysis. 
     
     
         45 . The method of  claim 44 , where the preloaded compounds are sterilized by sterile filtration. 
     
     
         46 . The method of  claim 34 , further comprising analyzing the drawn blood sample by a flow cytometer without further dilution and/or treatment. 
     
     
         47 . The method of  claim 34 , further comprising screening the drawn blood sample for HIV.