Apparatus, system and method for collecting a target material
This disclosure is directed to an apparatus, system and method for retrieving a target material from a sample. An enrichment agent may be added to a vessel that contains the sample for positive selection, or, in other words, to select or aid in selecting the target material from amongst the remainder of the sample. The enrichment agent may be, for example, immunomagnetic beads, buoyant beads, high-density beads, chemicals to change the density of the target material, or the like.
1. A method comprising the steps of:
providing a primary vessel comprising an open end and a suspension comprising a target material;
adding a positive enrichment reagent to the suspension for positive selection of the target material;
inserting a device comprising a cannula into the open end of the primary vessel, wherein the device extends upwardly from the open end of the primary vessel;
providing a processing vessel comprising
a first end comprising a resealable plug, and
an inner cavity;
mating the processing vessel with the device, wherein the cannula extends through the resealable plug of the processing vessel and accesses the inner cavity of processing vessel, and wherein the processing vessel extends upwardly from the device;
adding a displacement fluid comprising a density greater than a density of the target material to the inner cavity of the processing vessel; and
centrifuging the primary vessel, the device, the displacement fluid, and the processing vessel, wherein during centrifuging, the displacement fluid flows into the primary vessel via the device and displaces the target material from the primary vessel, which flows into the processing vessel through the device via the cannula.
2. The method of claim 1 , further comprising the step of:
separating the suspension into fractions prior to inserting step.
3. The method of claim 2 , further comprising the step of:
adding a first depletion agent to the suspension to remove a first sample fraction from the suspension or to change the density of at least a first non-target material relative to the density of the target material.
4. The method of claim 3 , further comprising the step of:
adding a second depletion agent to the suspension to remove a second sample fraction from the suspension or to change the density of at least a second non-target material relative to the density of the target material.
5. The method of claim 2 , further comprising the step of:
removing at least a portion of a non-target material from the primary vessel after the separating step and before the inserting step.
6. The method of claim 5 , further comprising the step of:
adding a clearing fluid to the primary vessel after the removing step, the clearing fluid having a density greater than at least the target material.
7. The method of claim 6 , further comprising the step of:
adding a float to the primary vessel prior to the separating step.
8. The method of claim 7 , further comprising the step of:
forming a first seal between the float and the primary vessel by clamping after the separating step, the removing step, or adding the clearing fluid.
9. The method of claim 8 , further comprising the step of:
centrifuging the primary vessel after forming the first seal.
10. The method of claim 9 , further comprising the step of:
forming a second seal between the float and the primary vessel by clamping after the centrifuging step performed after forming the first seal,
wherein the first seal is formed between a lower end of a main body of the float and an inner wall of the primary vessel, and
wherein the second seal is formed between an upper end of the main body of the float and the inner wall of the primary vessel.
11. The method of claim 1 , further comprising the step of:
forming a seal between an end of the device and an inner wall of the primary vessel to maintain a fluid-tight sealing engagement between the end of the device and inner wall of the primary vessel.
12. The method of claim 1 , wherein the displacement fluid is selected from the group consisting of: a solution of colloidal silica particles coated with polyvinylpyrrolidone, a polysaccharide solution, iodixanol, a liquid wax, an oil, a gas, olive oil, mineral oil, silicone oil, immersion oil, mineral oil, paraffin oil, silicon oil, fluorosilicone, perfluorodecalin, perfluoroperhydrophenanthrene, perfluorooctylbromide, ionic liquids, a polymer-based solution, a surfactant, a perfluoroketone, perfluorocyclopentanone, perfluorocyclohexanone, a fluorinated ketone, a hydrofluoroether, a hydrofluorocarbon, a perfluorocarbon, a perfluoropolyether, silicon, a silicon-based liquid, phenylmethyl siloxane, and combinations thereof.
13. The method of claim 1 , wherein the positive enrichment reagent is a solution of colloidal silica particles coated with polyvinylpyrrolidone, a polysaccharide solution, iodixanol, a complex, branch glucan, cesium chloride, sucrose, a sugar-based solution, a polymer solution, a multi-phase polymer solution, a tetrameric antibody complex, a particle, a nanoparticle, a metal-based compound, a metal complex, a lipid, or a sugar.
14. A system comprising:
a primary vessel comprising
an open end,
a suspension comprising a target material, and
a positive enrichment reagent to the suspension for positive selection of the target material;
a processing vessel comprising
a first end comprising a resealable plug,
an inner cavity, and
a displacement fluid comprising a density greater than a density of the target material, wherein the displacement fluid is located within the inner cavity;
a device at least partially located within the open end of the primary vessel, the device comprising a cannula,
wherein the cannula extends through the resealable plug of the processing vessel and accesses the inner cavity of the processing vessel, thereby mating the processing vessel and the device and fluidly connecting the primary vessel to the processing vessel,
wherein the device extends upwardly from the open end of the primary vessel, and
wherein the processing vessel extends upwardly from the device.
15. The system of claim 14 , the primary vessel further comprising a clearing fluid comprising a density greater than a density of at least the target material.
16. The system of claim 14 , further comprising:
a float located at a longitudinal position within the primary vessel.
17. The system of claim 16 , further comprising a sealing ring located circumferentially around the primary vessel at the same longitudinal position as at least a portion of the float within the primary vessel.
18. The system of claim 14 , further comprising a seal between an end of the device and an inner wall of the primary vessel to maintain a fluid-tight sealing engagement between the end of the device and inner wall of the primary vessel.
19. The system of claim 14 , wherein the displacement fluid is selected from the group consisting of: a solution of colloidal silica particles coated with polyvinylpyrrolidone, a polysaccharide solution, iodixanol, a liquid wax, an oil, a gas, olive oil, mineral oil, silicone oil, immersion oil, mineral oil, paraffin oil, silicon oil, fluorosilicone, perfluorodecalin, perfluoroperhydrophenanthrene, perfluorooctylbromide, ionic liquids, a polymer-based solution, a surfactant, a perfluoroketone, perfluorocyclopentanone, perfluorocyclohexanone, a fluorinated ketone, a hydrofluoroether, a hydrofluorocarbon, a perfluorocarbon, a perfluoropolyether, silicon, a silicon-based liquid, phenylmethyl siloxane, and combinations thereof.