CN106062170A

CN106062170A - Apparatus, system, and method for collecting a target material

Info

Publication number: CN106062170A
Application number: CN201580011850.4A
Authority: CN
Inventors: L·尤伦; D·E·坎普顿; J·J·诺德伯格; S·卡雷; J·E·伦特
Original assignee: Rarecyte Inc
Current assignee: Rarecyte Inc
Priority date: 2014-02-04
Filing date: 2015-01-30
Publication date: 2016-10-26
Anticipated expiration: 2035-01-30
Also published as: EP3102665A4; EP3102665A1; CN106062170B; CN111139175A; WO2015119863A1

Abstract

This disclosure is directed to an apparatus, system and method for retrieving a target material from a suspension. A system includes a plurality of processing vessels and a collector. The collector funnels portions of the target material from the suspension through a cannula and into the processing vessels. Sequential density fractionation is the division of a sample into fractions or of a fraction of a sample into sub-fractions by a step-wise or sequential process, such that each step or sequence results in the collection or separation of a different fraction or sub-fraction from the preceding and successive steps or sequences. In other words, sequential density fractionation provides individual sub-populations of a population or individual sub-sub- populations of a sub-population of a population through a series of steps.

Description

For collecting the instrument of target material, system and method

With cross reference to related applications

This application claims in the interests of U.S. Provisional Patent Application No. 61/935,457 that on February 4th, 2014 submits to, and The application is also the part continuation application of the U.S. Patent Application No. 14/495,449 in JIUYUE in 2014 submission on the 24th, described U.S. State's number of patent application 14/495,449 is the part of the U.S. Patent Application No. 14/090,337 in submission on November 26th, 2013 Continuation application, described U.S. Patent Application No. 14/090,337 requires in the interim Shen of United States Patent (USP) that on November 30th, 2012 submits to Please number 61/732,029；U.S. Provisional Patent Application No. 61/745,094 in December in 2012 submission on the 21st；In March, 2013 The U.S. Provisional Patent Application No. 61/791,883 submitted to for 15th；In the U.S. Provisional Patent Application No. that on May 1st, 2013 submits to 61/818,301；Interests with the U.S. Provisional Patent Application No. 61/869,866 submitted on August 26th, 2013；And this Shen The most also it is the part continuation application of U.S. Patent Application No. 14/266,939 submitted on May 1st, 2014, described United States Patent (USP) Application number 14/266,939 require in the U.S. Provisional Patent Application No. 61/818,301 that on May 1st, 2013 submits to, in 2013 The U.S. Provisional Patent Application No. 61/869,866 that August 26 submitted to, and interim in the United States Patent (USP) of submission on February 4th, 2014 The interests of application number 61/935,457.

Technical field

Present disclosure relates generally to fluid based on density separately, and particularly to reclaiming target material from suspension.

Background

Suspension generally includes and is difficult to detect, extract and separate the purpose material for analyzing.Such as, whole blood is that material exists Suspension in fluid.Billions of erythrocyte that material is included in the protein fluid of referred to as blood plasma and leukocyte and blood are little Plate.Whole blood is carried out routine examination by the existence with regard to aberrant biological or cell, and described aberrant biological or cell such as ovum, fetus is thin Born of the same parents, endotheliocyte, parasite, antibacterial and inflammatory cell and virus include HIV, cytomegalovirus, hepatitis C virus and EB Virus.At present, practitioner, researcher and those people with blood sample work attempt separately, separate and extract certain of peripheral blood sample A little components are used for checking.Comprise the following steps for analyzing the usual technology of blood sample: by blood film smear at microscope slide, and with Such mode dyeing film so that some component can be checked by bright-field microscopy art.

On the other hand, the purpose material occurred with extremely low concentration in suspension is difficult to (if if not impossible to) to be made Detect by many prior aries and analyze.Consider such as circulating tumor cell (" CTC "), its be from tumor depart from, Blood flow circulates, and the cancerous cell of the seed of the subsequent growth of the other tumor can being considered as in different tissues (i.e. turns Move).Accurately detection and analyze the ability of CTC be oncologist and cancer research person of special interest.But, CTC is with extremely low Number occurs in peripheral blood sample.Such as, the 7.5ml sample containing few peripheral blood sample to 5 CTC is considered as cancer Diagnosis and the treatment of patient are correlated with clinically.In other words, 5 CTC in detection 7.5ml blood sample are equivalent to detect about 100 1CTC in the background of hundred million erythrocyte and leukocyte, this uses blood film analysis extremely time-consuming, expensive and is difficult to.

Therefore, practitioner, researcher and those people with suspension work continue to look for the existence with regard to rare purpose material Or do not exist and accurately analyze the system and method for suspension.

Accompanying drawing describes

Figure 1A-1B shows example catcher.

Fig. 2 A-2B shows example catcher.

Fig. 2 C-2D shows example catcher.

Fig. 3 A-3B shows example catcher-process vessel system.

Fig. 4 A-4B shows example catcher-shelter (canopy) system.

Fig. 5 A-5B shows example seal ring.

Fig. 5 C-5D shows example seal ring.

Fig. 5 E-5F shows example seal ring.

Fig. 5 G shows example seal ring.

Fig. 6 shows the flow chart of the exemplary method for reclaiming target material.

Fig. 7 A-7B shows example float and Primary containers system.

Fig. 8 shows and experiences separate example float based on density and Primary containers system.

Fig. 9 shows and forms the example seal ring and example float and Primary containers system sealed.

Figure 10 A-10G shows the example system reclaiming target material.

Describe in detail

Present disclosure relates to reclaim the instrument of target material, system and method from suspension.System includes multiple Process vessel and catcher.The part of the target material from suspension funnel is injected in process vessel by catcher.Sequential close Degree classification is by progressively or sample is divided into fraction or sample fraction is divided into subfraction by sequential process so that each step or Order causes from previous and sequential step or the different fractions of order or the collection of subfraction or separates.In other words, sequential close Degree classification provides each subgroup or each sub-subgroup (sub-sub-of colony's subgroup of colony by series of steps population)。

Catcher

Figure 1A shows the isometric view of catcher 100.Figure 1B shows the collection obtained along the line I-I shown in Figure 1A The viewgraph of cross-section of device 100.Chain-dotted line 102 represents the center of catcher 100 or the highest axis of symmetry.Catcher 100 can have Such size and dimension, to fit in containing in the Primary containers that maybe can accommodate suspension, described suspension bag under a cloud Include target material.Catcher 100 by from suspension target material funnel by intubate 106 injections and enter process vessel (not Illustrate) in, to be positioned at chamber 108.Catcher 100 includes main body 104, and it includes first end 110 and the second end 112.Close Envelope can be formed between the second end 112 and the inwall of Primary containers, with before centrifugation, centrifugal period and centrifugal after maintain stream The close sealing engagement of body, and suppress any part of suspension to be positioned at inwall and the main body of catcher 100 of Primary containers Between 104, or flow between the inwall and the main body 104 of catcher 100 of Primary containers.Sealing can by following come shape Become: interference engagement, grease (such as vacuum grease), binding agent, epoxy resin, by bonding (such as passing through hot adhesion), By welding (such as passing through ultrasonic bonding), by clamping (such as with ring or fixture), fit in the second end 112 and main Insert (such as O or the collar) between the inwall of container etc..Main body 104 can be any suitable shape, including but It is not limited to cylinder, triangle, square, rectangle etc..Catcher 100 also includes internal funnel 114, and it is concave-shaped openings. Funnel 114 can be tapered towards intubating 106 from the second end 112.Funnel 114 is by from the target less than the second end 112 In material is directed to intubate 106, described in intubate 106 and be connected with the summit of funnel 114 and be in fluid communication.The summit tool of funnel 114 There is the diameter less than the oral area of funnel 114.Funnel 114 is formed by tapered wall, and described tapered wall is permissible Be straight, curve, arch etc..Funnel 114 can be any suitable shape, includes but not limited to tubular, spherical, round Top shape, cone, rectangle, pyramid etc..Additionally, the most external diameter of funnel 114 or edge can be with Primary containers Inwall connects continuously or constant contact (i.e. flushing) so that the second end 112 of catcher 100 and Primary containers inwall it Between there is not Dead space.

Intubate 106 such as pipe or pins and include but not limited to non-coring pin, extend from the summit of funnel 114 and enter chamber 108 In.In the example in fig 1, chamber 108 is to extend into the concave-shaped openings in main body 104 from first end 110, and can accept And support process vessel (not shown).Chamber 108 can be any suitable degree of depth, to accept and to support that process vessel (does not shows Go out).Intubate 106 can extend in any appropriately distance entrance chamber 108, in order to penetrate the base portion of process vessel (not shown), Or in insertion process vessel.Intubate 106 and can include flat tip, chamfered tip, sharp tip or cone point.Additionally, chamber 108 can be any suitable shape, includes but not limited to tubular, spherical, cheese, cone, rectangle, pyramid etc. Deng.Chamber 108 can be threaded, to engage the threaded section (not shown) of process vessel.

Catcher 100 can also include keeper (not shown), to stop catcher 100 to slide relative to Primary containers, So that catcher 100 is maintained at the predetermined height in Primary containers.Keeper (not shown) can be from first end 110 The shoulder, the fixture that radially extend, extend beyond the annular projection of the circumference of cylinder-shaped body 104, retainer etc..

Fig. 2 A shows the isometric view of catcher 200.Fig. 2 B shows the receipts obtained along the line II-II shown in Fig. 2 A The viewgraph of cross-section of storage 200.Chain-dotted line 202 represents the center of catcher 200 or the highest axis of symmetry.Catcher 200 is similar to Catcher 100, except catcher 200 includes the main body 204 more elongated than the main body of catcher 100, in order to accommodates process vessel Outside the greater part of (not shown).Main body 204 includes first end 206 and the second end 208.Sealing can be at the second end Formed between 208 and the inwall of Primary containers, with before centrifugation, centrifugal period and centrifugal after maintain the sealing of fluid-tight to connect Close, and suppress any part of suspension to flow between the inwall and the main body 204 of catcher 200 of Primary containers.Seal Can be formed by following: interference engagement, grease (such as vacuum grease), binding agent, epoxy resin, bonding are (such as Hot adhesion), by welding (such as passing through ultrasonic bonding), clamping (such as with ring or fixture), fit in the second end 208 and Insert (such as O or the collar) between the inwall of Primary containers etc..

First end 206 includes at least one of chamber 212 a size of accepting and accommodating process vessel (not shown).Chamber 212 can have taper or the stepped bottom end 220 that process vessel (not shown) can be positioned on.First end 206 can also include at least one otch (cut-out) 210, to allow suitably grasping for inserting of process vessel (not shown) Enter and take out.Catcher 200 by the target material funnel from suspension by intubating at 214 injection the second ends 208 in In portion's funnel 222, and enter the process vessel (not shown) being positioned at chamber 212.Intubate 214 can be placed on frame 224, make The internal holes of 214 must be intubated flush, as shown in Figure 2 B with the inwall of funnel 222.

Catcher 200 can include shoulder 216, and described shoulder 216 circumference around main body 204 extends.Shoulder 216 is permissible It is longer than the internal diameter of Primary containers, in order to be placed in the open end of Primary containers, and to Primary containers and shoulder 216 After outside applying locking ring (not shown), to suppress the catcher 200 movement relative to Primary containers.Locking ring (not shown) edge Shoulder 216 applies pressure to Primary containers.Locking ring can be two rings, Primary containers whole circumference wrap up one Loop or at the loop wrapped up less than whole circumference of Primary containers, such as 1/2nd (1/2), 5/8ths (5/ 8), 2/3rds (2/3), 3/4ths (3/4), 7/8ths (7/8) etc..Alternatively, shoulder 216 can fit in mainly In container.Alternatively, shoulder 216 can be fixture so that shoulder 216 can include that Primary containers may be inserted into its interior card Lock, to suppress the catcher 200 movement relative to Primary containers.Alternatively, shoulder 216 can be with being internally formed of Primary containers Interference engagement, sealing ring can be placed in about.

As shown in Figure 2 A, catcher 200 can include at least one window 218, to be entered by the inwall of main body 204 (access) chamber 212.At least one window 218 allows operator to confirm process vessel (not shown) suitably putting in chamber 212 Put.At least one window 218 also allows for from intubating removing fluid 214, to flow out catcher 200 and to enter at catcher 200 And formed between Primary containers (not shown), and above the sealing between the second end 208 and the inwall of Primary containers In space.

Fig. 2 C shows the isometric view of catcher 230.Fig. 2 D shows along the line III-III acquisition shown in Fig. 2 C The viewgraph of cross-section of catcher 230.Catcher 230 is similar to catcher 200, except catcher 230 includes main body 238, described Opening in prolongation 234 that main body 238 includes extending outwardly away from first end 232 and at least temporarily sealing prolongation 234 The lid 236 of 240.Opening 240 can be in fluid communication with chamber 212 at first end 232.Lid 236 can be removable, can penetrate With reclosable (such as hinged cover), maybe can penetrate and the most reclosable (such as foil lid).Prolongation 234 can have Accept the size of lid 236 when penetrating so that a part for lid 236 cannot extend in chamber 212 at first end 232. It should be noted that catcher 230 does not include at least one otch 210.

Main body can be made up of various different materials, and described various different materials include but not limited to pottery；Metal；Organic Or inorganic material；With plastic material such as polyformaldehydePolystyrene, acronitrile-butadiene-styrene (" ABS ") copolymer, aromatic polycarbonate, aromatic polyester, carboxymethyl cellulose, ethyl cellulose, ethylene vinyl acetate are common Polymers, nylon, polyacetals, poly-acetas, polyacrylonitrile and other nitrile resins, acrylonitrile-vinyl chloride copolymer, polyamide, Aromatic polyamides (" aramid "), polyamide-imides, polyarylate, polyarylene oxide, poly arylidene thio-ester, polyarylsufone, Polybenzimidazoles, polybutylene terephthalate (PBT), Merlon, polyester, polyesterimide, polyether sulfone, Polyetherimide, poly- Ether ketone, polyether-ether-ketone, polyethylene terephthalate, polyimides, polymethacrylates, polyolefin (such as polyethylene, poly-third Alkene), polyallomer (polyallomers), poly-diazole, Parylene, polyphenylene oxide (PPO), MODIFIED PP O, polystyrene, Polysulfones, fluoropolymer such as politef, polyurethanes, polyvinylacetate, polyvinyl alcohol, polyvinyl halogenation Thing such as polrvinyl chloride, polyvinyl chloride-acetate copolymer, polyvinylpyrrolidone, polyvinylidene chloride, extraordinary polymerization Thing, polystyrene, Merlon, polypropylene, acrylonitrilebutadienestyrene copolymer, butyl rubber, propylene diene list Body；And combinations thereof.

Intubating and can be made up of various different materials, described various different materials include but not limited to pottery；Metal；Organic Or inorganic material；With plastic material such as polypropylene, acrylic acid, Merlon etc.；And combinations thereof.Intubate and can have along inserting The tip of the pipe longitudinal axis.

Catcher-process vessel system

Fig. 3 A shows example catcher 200 and the decomposition view of process vessel 302.Fig. 3 B shows along shown in Fig. 3 A Line IV-IV obtain, at the first end 206 collecting container 200 cross section of the process vessel 302 in insertion chamber 212 View.Catcher 200 and process vessel 302 form catcher-process vessel system 300.Process vessel 302 can be Eppendorf pipe, syringe or test tube, and there is Guan Bi end 304 and open end 306.Open end 306 has acceptance The size of cap 308.Cap 308 can be made up of resealable rubber or other suitable resealable materials, described resealable Material can pierce through repeatedly by pin or other sharp instrument, with the content of storage in process vessel 302 inside, and works as Pin or utensil seal when removing again.Alternatively, process vessel 302 can also have two open end of the size accepting cap. Process vessel 302 can have the taper geometry widened towards open end 306 or narrow；Process vessel 302 can have There is the geometry of substantial cylindrical；Or, process vessel 302 can have the geometric form of substantial cylindrical in the first section Shape and in the second section the geometry of cone, wherein the first section and the second section are connected to each other and continuously.Although adding At least one section of chemical container 302 has circular cross section, but in other embodiments, at least one section can have Ellipse, square, triangle, rectangle, octagon or any other suitable shape of cross section.Process vessel 302 can be by Transparent, translucent, opaque or semipermeable materials such as plastics or another kind of suitable material composition.Process vessel includes center Axle 314, when inserting in chamber 212, described central shaft 314 is coaxial with the central shaft 202 of catcher 200.Process vessel 302 also may be used To be included in the plug 310 at Guan Bi end 304, to allow the introducing of target material or to make target material exchange with displacement fluid 312.Close It can be threaded for closing end 304, to provide and the threadeding of the threaded chamber 212 of catcher 200.Process vessel 302 Can be made up of glass, plastics or other suitable materials.

Plug 310 can be made up of resealable rubber or other suitable resealable materials, described resealable material Repeatedly can pierce through by pin or other sharp instrument, to introduce close to the content within process vessel 302 or permission content In process vessel 302, and when pin or utensil seal when removing again.Plug 310 may be inserted in process vessel 302 so that seals Such as maintained between plug 310 and process vessel 302 by interference engagement.Alternatively, plug 310 can use heating liquid rubber Being formed in the Guan Bi end 304 of process vessel 302, described heating liquid rubber can shape when heating or being hot and work as rubber Harden during cooling.Binding agent may be used for being attached to plug 310 inwall of process vessel, can be bonding based on polymer Agent, epoxy resin, contact adhesive or for boning or producing any other suitable material of hot adhesion.Alternatively, plug 310 can be mapped in process vessel 302.Alternatively, plug 310 can be with process vessel 302 hot adhesion.

In the example of Fig. 3 B, intubating 214 and have cone point, it penetrates plug 310 and extends into process vessel 302 Inner chamber, wherein intubates the handle of 214 and does not extends into the inner chamber of process vessel 302.As described in more detail below, processing The inner chamber of container 302 accommodates target material.Intubate 214 can be covered by resealable sleeve (not shown), to stop target material Flow out, unless process vessel 302 intubates 214 degree of depth just penetrating process vessel 302 to allowing in chamber 212.Resealable Sleeve (not shown) covers and intubates 214, is spring, can be intubated 214 and penetrate, and is repeated thorn by withstanding Wear and still maintain the elastomeric material of sealing to make simultaneously.

As shown in Fig. 3 A-3B, before inserting in catcher 200, process vessel 302 can be mounted with displacement fluid 312.Displacement fluid 312 replaces target material so that include target material Primary containers when catcher 200 and process vessel 302 insert In (not shown), and when catcher, process vessel and Primary containers experience are centrifugal, displacement fluid 312 flows out process vessel 302 And in entrance Primary containers, and such as replace (that is, making material be lifted up) by buoyancy by displacement, promote target material to lead to Cross and intubate 214 and enter in process vessel 302.

Displacement fluid 312 has the density bigger than the density of the target material of suspension, and (density can be more than suspension level The subset divided or the density of all suspension fraction), and be inert for suspension material.Displacement fluid 312 is suspending Fluid can be miscible or the most miscible.The suitably example of displacement fluid includes but not limited to be coated with polyethylene The colloidal silica particle solution (such as Percoll) of ketopyrrolidine, polysaccharide solution (such as Ficoll), iodixanol are (such as OptiPrep), organic solvent, liquid wax, oil, gas and combinations thereof；Olive oil, mineral oil, silicone oil, immersion oil, mineral oil, Paraffin oil, silicone oil, fluorosilicone, perfluorodecalin, perfluor perhydro phenanthrene, perfluoroctylbromide and combinations thereof；Organic solvent is such as 1,4-dioxane, acetonitrile, ethyl acetate, the tert-butyl alcohol, Ketohexamethylene, dichloromethane, tert-pentyl alcohol, methyl tertiary butyl ether(MTBE), butyl acetate, Hexanol, Nitrobenzol, toluene, capryl alcohol, octane, Allyl carbonate, sulfolane and ionic liquid；Solution based on polymer；Surface Activating agent；Perfluor ketone such as perfluorocyclopentanone and perfluorocyclohexanone, fluorinated ketone, hydrofluoroether, hydrogen fluorohydrocarbon, perfluocarbon, perfluor gather Ether, silicon and liquid based on silicon such as phenylmethylsiloxane；And combinations thereof.

Process vessel 302 can also include process solutions (not shown), with when target material enters process vessel 302, real The now conversion to target material.Process solutions (not shown) can be preservative, cell adhesion solution, dyestuff etc..With displacement stream Body 312 is different, and great majority (and if not all) process solutions (not shown) is retained in process vessel 302 after centrifugal, from And in one way or another way (i.e. anticorrosion, increase adhesion characteristics etc.) realizes the conversion to target material.Process solutions (not shown) can introduce as the liquid comprised in liquid or shell.Shell can be dissolved in aqueous solution rather than displacement fluid In 312 (such as gel cap)；Or shell can be frangible so that when process vessel 302 vibrates in turbine mixer, outward Shell ruptures.Further, it is possible to use exceed a kind of process solutions.

Process vessel 302 can include flexible cap, and it can be promoted will to be assigned to substrate example by its predetermined On microscope slide or orifice plate.Cap 308 can be flexible, or cap 308 can be removed, and flexible cap is inserted open end In 306.Alternatively, process vessel 302 can be attached to allotter (i.e. after accumulation target material), maybe can include allotter, The target material of predetermined can be assigned to another kind of substrate such as microscope slide from process vessel 302 by described allotter On.Allotter can repeat to pierce through the material in resealable cap 308, or compression process container 302, predetermined with extraction and distribution The target of volume is expected in substrate.Alternatively, cap 308 can be removed, and allotter (not shown) is directly inserted into and adds In chemical container 302, to distribute buffy coat-process solutions mixture.

Catcher-shelter system

Fig. 4 A shows example catcher 200 and the decomposition view of shelter 402.Fig. 4 B shows along the line shown in Fig. 4 A V-V obtains, the viewgraph of cross-section of the process vessel 402 in the chamber 212 of insertion catcher 200.Catcher 200 and shelter 402 Form catcher-shelter system 400.Shelter 402 is similar to process vessel 302, except shelter have the second open end 404 it Outward.When catcher-shelter system 400 is inserted in Primary containers, the such as part of some fluids in Primary containers suspends Liquid, a part of suspension fraction, part cleaning fluid etc., can be by intubating 214 discharges.Shelter 402 suppression can be led to Cross the opening that a part of fluid in the Primary containers intubating 214 discharges escapes from the first end 206 of catcher 200.Pass through The discharge fluid that shelter 402 blocks flows out the second open end 404, and flows out window 218.Dotted line 406 display is passed through when fluid Intubate fluid flowing during 214 discharge, and retained by shelter 402.

Alternatively, when using catcher 230, lid 236 suppression of catcher 230 can be by intubating the master of 214 discharges Wanting a part of fluid in container to escape from the opening of first end 206 of catcher 200, its mode is similar to that of shelter 402 Kind.

Sealing ring

Fig. 5 A shows the isometric view of sealing ring 500.Fig. 5 B shows the lower view from top of sealing ring 500.Dot-dash Line 502 represents the center of sealing ring 500 or the highest axis of symmetry.Sealing ring 500 includes inwall 504, outer wall 506 and chamber 508.At figure In 5B, R_IWRepresent from the center of sealing ring 500 to the radial distance of inwall 504, and R_OWRepresent from the center of sealing ring 500 Radial distance to outer wall 506.Sealing ring 500 is configured to fit in around Primary containers such as pipe.It is main that chamber 508 has acceptance The size and shape of container.Sealing ring 500 can screw so that applies pilot seal ring 500 by circumference around outer wall 506 The substantially uniform radial force of central shaft 502, the radial force such as produced by fixture, the size in chamber 508 and inwall and The radius of outer wall 504 and 506 reduces.When sealing ring 500 screws around Primary containers, put on the uniform of sealing ring 500 Power puts on Primary containers, thus promotes Primary containers to shrink.When radial force is removed from sealing ring 500, sealing ring 500 is being led Container surroundings is wanted to keep screwing and being in tension force.

Sealing ring can be any shape, includes but not limited to circle, triangle or polyhedron.Fig. 5 C shows sealing ring The isometric view of 510.Fig. 5 D shows the lower view from top of sealing ring 510.Sealing ring 510 is similar to sealing ring 500, except Sealing ring 510 is outside polyhedron.Chain-dotted line 512 represents the center of sealing ring 510 or the highest axis of symmetry.Sealing ring 510 includes Inwall 514, outer wall 516 and chamber 518.Sealing ring can be by metal such as pyrite, polymer or combinations thereof.

Alternatively, as shown in fig. 5e, sealing ring 520 can be made up of piezoelectric.Fig. 5 F shows sealing ring 520 The lower view from top.Chain-dotted line 522 represents the center of sealing ring 520 or the highest axis of symmetry.Sealing ring 520 can be led via first Line 524 and the second wire 526 are attached to potential source 528, such as battery.Potential source 528 produces and promotes sealing ring 520 to tighten (i.e. Sealing ring 520 radius reduce) mechanical stress.Sealing ring 520 includes inwall 530, outer wall 532 and chamber 534.In Fig. 5 F, R_IW Represent from the center of sealing ring 520 to the radial distance of inwall 530, and R_OWRepresent from the center of sealing ring 520 to outer wall 532 Radial distance.Alternatively, sealing ring 520 may be at natural tight condition.When an electrical potential is applied, sealing ring 520 is expanded.Standby Selection of land, a part for sealing ring can be made up of piezoelectric so that piezoelectric serves as driver, to promote its of sealing ring He partly tightens and Primary containers is applied substantially uniform circumferential pressure, so that Primary containers shrinks to form sealing.

Fig. 5 G shows the isometric view of sealing ring 540.Sealing ring includes guiding mechanism 548, to adjust internal diameter R_ID.Can roll over Folded ring includes first end 542 and the second end 546, described first end 542 and the second end 546 by band portion 544 even Connect.First end 542 and the second end 546 include the complementary portion of guiding mechanism 548.Guiding mechanism 548 includes but not limited to Ratchet, tongue-and-groove, ratchet etc..

Sealing ring can also include temperature-sensitive element, such as heater wire.Temperature-sensitive element can soften Primary containers for shrinking. Alternatively, temperature-sensitive element can melt Primary containers, the sealing more adhered to offer.Alternatively, temperature-sensitive element can promote close Seal ring is compressed, thus is formed between Primary containers and float and seal.

Sequential density step-procedure

Sequential density classification is by progressively or sample is divided into fraction by sequential process or sample fraction is divided into subfraction, makes Obtain each step or order causes from previous and sequential step or the different fractions of order or the collection of subfraction or separates.Change Yan Zhi, sequential density classification provides each subgroup or each sub-subgroup of colony's subgroup of colony by series of steps.Example As, buffy coat is the fraction of whole blood sample.Buffy coat fraction can further decompose into subfraction, including but do not limit In reticulocyte, granulocyte, lymphocyte/mononuclear cell and platelet.These subfractions can be by performing sequential density Each acquisition of classification.

For convenience, the method is described with reference to the example flyout liquid of anticoagulated whole blood.But method described below does not has So limit in being directed at its range of application.In practice, the method may be used for any kind of suspension.Example As, sample suspension can be urine, blood, bone marrow, capsule liquid, ascites, feces, seminal fluid, cerebrospinal fluid, nipple aspirate fluid, saliva Liquid, amniotic fluid, vaginal secretions, Mucosal secretions, aqueous humor, vitreous humor, vomitus and any other physiological fluid or half are admittedly Body.It should also be understood that target material can be the fraction of sample suspension, such as buffy coat, cell such as ovum, tire Youngster's material (such as trophoderm, erythroblast, FE, fetal leukocytes, foetal DNA, fetal rna etc.) or circulation Tumor cell (" CTC "), circulating endothelial cells, immunocyte (i.e. initial or memory B cell or initial or memory T cell), Vesicle, liposome, protein, nucleic acid, biomolecule, there is the microscopic units naturally occurring or manually preparing of closing membrane, post It is infested that (such as spirillum such as causes the B. burgdorferi (Borrelia burgdorferi) of Lyme disease；Malaria (malayria) derivant), microorganism, virus or inflammatory cell.Alternatively, sample can be that biosolids is such as organized, Before or after adding Primary containers, described biosolids is the most such as decomposed by collagenase.

Fig. 6 shows that the sequential density classification of use is for reclaiming the flow chart of the exemplary method of target material.At square frame 602 In, it is thus achieved that suspension such as anticoagulated whole blood.In square frame 604, whole blood is added in Primary containers such as test tube.Float can also Add in Primary containers.For convenience, the method is described for float, but methods described below is not directed at Its application so limits, and can perform without float.

Fig. 7 A shows example Primary containers and the isometric view of float system 700.System 700 includes Primary containers 702 With the float 704 being suspended in whole blood 706.In the example of Fig. 7 A, Primary containers 702 has circular cross section, the first opening End 710 and the second Guan Bi end 708.Open end 710 has the size accepting cap 712.Primary containers can also have and connects By two open end of the size of cap, such as the example pipe shown in Fig. 7 B and discerptible float system 720.System 720 class Being similar to system 700, in addition to Primary containers 702 replaces with Primary containers 722, described Primary containers 722 includes being configured to point Do not accept two open end 724 and 726 of cap 712 and cap 728.Primary containers 702 and 722 has the geometry of substantial cylindrical Shape, widens respectively towards open end 710 and 724 it is also possible to have, narrows or the taper geometry of a combination thereof.Although Primary containers 702 and 722 has circular cross section, but in other embodiments, Primary containers 702 and 722 can have ellipse Any other suitable shape of cross section of circle, square, triangle, rectangle, octagon or substantially extension length.Main Want the container 702 and 722 can be by transparent, translucent, opaque or semipermeable materials such as plastics or another kind of suitable material group Become.Primary containers 702 and 722 each includes central shaft 718 and 730 respectively.Primary containers 702 is additionally may included in Guan Bi end Barrier film 714 at 708, as visible in zoomed-in view 716, to allow fluid, suspension or the removal of suspension fraction, no matter Be with syringe, pump, by drain etc..Primary containers 702 can have inwall and the first diameter.

Barrier film 714 can be made up of resealable rubber or other suitable resealable materials, described resealable material Material can pierce through repeatedly by pin or other sharp instrument, to enter the content within Primary containers 702, and when pin or utensil Seal again during removal.Barrier film 714 may be inserted in Primary containers 702 so that seals and such as maintains barrier film by interference engagement Between 714 and Primary containers 702.Alternatively, barrier film 714 can use heating liquid rubber in the opening and/or bottom of pipe Being formed in portion, described heating liquid rubber can shape when heating or being hot and harden when rubber cools down.Binding agent can be used In barrier film 714 being attached to the wall within opening and pipe, and can be binding agent based on polymer, epoxy resin, contact Binding agent or for by rubber bound to plastics or produce hot adhesion any other suitable material.Alternatively, barrier film 714 Can be with Primary containers 702 hot adhesion.

Float 704 includes main body, two tear drop shape end caps and is radially spaced in main body and the support of axial orientation Component.Alternatively, float 704 can not include any supporting member.Alternatively, float 704 can include supporting member, and it is not Engage the inwall of Primary containers 702.

In alternative embodiment, supporting member number, supporting member interval and support member thickness can be each independent Ground changes.Supporting member can also be destroyed or segmentation.Main body is dimensioned such that, to have less than Primary containers 702 The external diameter of internal diameter, thus the fluid retention passage being limited between the outer surface of main body and the inwall of Primary containers 702.Support structure Body surfaces between part can be smooth, bending or have another kind of suitably geometry.Supporting member and main body Can be single structure, can be maybe separate structure.

Embodiment includes the other kinds of geometry shape for float end cap.Top end cap can be tear drop shape, Cheese, cone or any other suitable shape.Bottom end cap can be tear drop shape, cheese, conical or any its The shape that he is suitable.In other embodiments, the main body of float 704 can include that various different supporting construction is for separately sample Product, support tube wall or guided around float by suspension in centrifugal period.Embodiment is not intended to be limited to these examples Son.Main body can include the many projections providing the support for pipe.In alternative embodiment, protrusions number and pattern are permissible Change.Main body can include single continuous print helical structure or shoulder, and it is spiral around main body, produces helical duct.At it In his embodiment, spiral shoulder can be mellow and full or broken or segmentation, to allow adjacent at spiral shoulder of fluid Flow between corner.In various embodiments, spiral shoulder interval and rib thickness can be changed independently.Another embodiment party In case, the supporting member that main body can include radially from the body and circumference extends around main body.In another embodiment, Supporting member can be taper.

Float 704 can be made up of various different materials, and described various different materials include but not limited to metal；Organic or Inorganic material；Iron content plastics；Sintering metal；The metal of machining；Plastic material and combinations thereof.Primary containers 702 can have There are inwall and the first diameter.Float 704 can be by captured in being an interference fit in Primary containers 702 so that when centrifugal, pipe Inwall expansion is to allow moving axially of float 704.When centrifugal stopping, inwall rolls back the first diameter, joins with induced interference Close.Alternatively, inwall can not be expanded, and interference engagement can occur without between float 704 and Primary containers 702, makes Float before centrifugation, centrifugal period or centrifugal after move freely in pipe.The end cap of float can pass through machining, injection Shaping, additive technology etc. are fabricated to a part for main body, thus become a single structure；Or, adaptive by pressing, Binding agent, screw, any other the suitable method at least two panels connected together by it or a combination thereof, end cap can couple To main body.

Cap 712 can be made up of various different materials, and described various different materials include but not limited to organic or inorganic material Material；Plastic material；And combinations thereof.

Returning to Fig. 6, in block 606, Primary containers, float and whole blood experience are such as by centrifugal based on density point Open, thus allow whole blood to be divided into fraction based on density based on density along the axial location of pipe.Fig. 8 show experience the most logical Cross centrifugal separate Primary containers based on density and the isometric view of float system 700.Assume that such as centrifugal whole blood includes Three fraction.For convenience, three fraction include blood plasma, buffy coat and erythrocyte.But, when another kind of suspension warp When going through centrifugal, can there is more, less or equal number of fraction, each fraction has different densities.As shown in Figure 8, Suspension experience is divided into the axially spaced of three fraction based on density along the length of pipe, and wherein erythrocyte 803 is positioned at bottom, blood plasma 801 are positioned at top, and buffy coat 802 is positioned between the two.Float 704 can have any suitable density, with in level Sedimentation in/mono-.The density of float 704 can so select so that float 704 is expanded, and buffy coat 802 is at float Between main body and the inwall of Primary containers.Buffy coat 802 can be trapped in the district between float 704 and Primary containers 702 In territory.

At least one divides fluid (delineation fluid) (not shown) and is provided for target material and at target Being spaced further apart between any non-target material of material above and or below.At least one divides fluid (not shown) can have There is the density more than or less than target material.Such as, when expectation is spaced further apart buffy coat 802 and erythrocyte 803, divide Fluid can have more than buffy coat 802 with less than the density of erythrocyte 803.At least one divides fluid (not shown) can Being miscible with suspension flow physical ability or the most miscible, and it is inert for suspension material.At least one divides fluid (not shown) may be provided for wherein sealing the region of Primary containers 702 because buffy coat 802 and erythrocyte 803 it Between there is bigger division and separately.Regardless of whether use float that at least one all can be used to divide fluid (not shown).Close The suitable example dividing fluid includes but not limited to be coated with the colloidal silica particle solution of polyvinylpyrrolidone (such as Percoll), polysaccharide solution (such as Ficoll), iodixanol (such as OptiPrep), cesium chloride, sucrose, based on sugar molten Liquid, solution based on polymer, surfactant, organic solvent, liquid wax, oil, gas and combinations thereof；Olive oil, mineral oil, Silicone oil, immersion oil, mineral oil, paraffin oil, silicone oil, fluorosilicone, perfluorodecalin, perfluor perhydro are luxuriant and rich with fragrance, PFO bromide Thing and combinations thereof；Organic solvent such as 1,4-dioxane, acetonitrile, ethyl acetate, the tert-butyl alcohol, Ketohexamethylene, dichloromethane, tert-pentyl alcohol, Methyl tertiary butyl ether(MTBE), butyl acetate, hexanol, Nitrobenzol, toluene, capryl alcohol, octane, Allyl carbonate, sulfolane and ionic liquid； Solution based on polymer；Surfactant；Perfluor ketone such as perfluorocyclopentanone and perfluorocyclohexanone, fluorinated ketone, hydrofluoroether, hydrogen Fluorohydrocarbon, perfluocarbon, PFPE, silicon and liquid based on silicon such as phenylmethylsiloxane；And combinations thereof.

Fig. 9 shows the sealing being formed as stoping fluid to move up or down in Primary containers.Seal also suppression floating Son is mobile.Sealing ring 500 applies circumference or radial force to Primary containers 702, thus promotes Primary containers 702 for float 704 Fold inward.Zoomed-in view 902 shows the sealing ring 500 tightened around float and Primary containers system 700.Have been placed in blood The sealing ring 500 of the interface of heavy buffycoat 802 and erythrocyte 803 promotes Primary containers 702 fold inward, until being sealed in master Want to be formed between container 702 and float 704.The outer wall of sealing ring 500 can flush with the outer wall of Primary containers 702；Sealing ring The outer wall of 500 can extend across the outer wall of Primary containers 702；Or, the outer wall of Primary containers 702 can extend across sealing The outer wall of ring 500.Sealing ring 500 keeps tightening to maintain sealing, and this stops fluid to be moved across sealing in any direction.Seal Ring 500 may also stay at tension state.Alternatively, sealing ring 500 can excessively be tightened, and removes subsequently sealing ring 500 power applied.Sealing ring 500 can slightly be expanded, although still remaining retracted.

In order to apply sealing ring 500 and thus form sealing, fixture may be used for the central shaft for Primary containers 702 Power circumference put on sealing ring 500 and float and Primary containers system 700.In float and Primary containers system 700 After going through such as by being centrifuged based on sealing separating, sealing ring 500 is placed in around float and Primary containers system 700.Seal Ring 500 and float and Primary containers system 700 are subsequently placed in fixture.Fixture can include that frame is with for Primary containers 702 Support sealing ring 500.The operation of fixture can be maybe can manually performing of automatization.Alternatively, fixture can form float Sealing between 704 and Primary containers 702, and without including sealing ring 500.Alternatively, ultrasonic bonding is such as passed through；Or it is logical Crossing applying heat or thermograde so that Primary containers 702 and float 704 deform and/or melted, sealing can be float 704 and master Want to be formed between container 702.For convenience, the method is described for the sealing between float and Primary containers, but Method described below is not directed in its application so limiting, and can perform without sealing.

When the operation of fixture is automatization, electromotor promote chuck include chuck refer to or pressure member translation, with promote Make the compression that chuck refers to.Electromotor can be attached to chuck or pressure structure by handle such as cam handle and one or more gear Part.Substrate engages and accommodates object.When chuck is driven by electromotor, pressure member keeps static.When pressure member passes through When electromotor drives, chuck keeps static.Fixture can include disconnector, in order to promotes pressure member to skid off chuck and refers to 904, from And remove clamping force.

Alternatively, fixture can be but not limited to collet fixture, o-ring, pipe clamp, hose clamp, spring perch, belt Fixture or tie such as band.Fixture can use without sealing ring, to provide the sealing between float and pipe.

As shown in FIG. 10A, blood plasma 801 can be such as by moving liquid, aspirating, topple over etc. and to take from Primary containers 702 Go out.Returning to Fig. 6, in block 608, cleaning fluid can add in Primary containers together with catcher-shelter system.Figure 10B-10C shows the cleaning fluid 1002 added in Primary containers 702, and it is close that it has more than at least buffy coat 802 Degree (that is, for example, it is possible to have more than buffy coat but less than erythrocytic density, maybe can have more than buffy coat Density with erythrocyte).Alternatively, during blood plasma 801 can be retained in Primary containers 702.When blood plasma 801 is retained in mainly Time in container 702, the density of blood plasma 801 can such as pass through iodixanol or any suitable substance change, close to change fraction Degree, thus serve as cleaning fluid.Therefore, when during blood plasma 801 is retained in Primary containers 702 and when density changes, can be not required to Fluid to be cleaned.

As shown in fig. 10d, catcher-shelter system 400 is subsequently added in Primary containers 702.The second of catcher 200 End 208 forms sealing 1008 with the inwall of Primary containers 702, with stop fluid before centrifugation, centrifugal period and centrifugal after Catcher 200 surrounding flows.Seal 1008 to be formed between the second end 208 and the inwall of Primary containers, with centrifugal Before, centrifugal period and centrifugal after maintain the sealing engagement of fluid-tight, and suppress any part of suspension to be positioned at main appearance Between inside and the main body 204 of catcher 200 of device, or flow between the main body 204 of the inside of Primary containers and catcher 200 Dynamic.Sealing can be formed by following: interference engagement, grease (such as vacuum grease), binding agent, epoxy resin, heat Bonding, ultrasonic bonding, clamping (such as with ring or fixture), fit between the second end 208 and the inwall of Primary containers Insert etc..Locking ring 1004 can be placed in the shoulder 216 of catcher 200 and Primary containers 702 open end 710 it On, to suppress the catcher 200 translation relative to Primary containers 702.When inserting catcher-shelter system 400, Primary containers A part for cleaning fluid 1002 in 702 by intubating 214 discharges, and can be stopped by shelter 402.Such as VI-VI along the line Obtain by seen by the dotted line 406 in zoomed-in view 1006, the fluid of discharge can be flowed out by window 218 and enter to be led In wanting container 702, although above the sealing being maintained between the inwall of the second end 208 and Primary containers 702.

In square frame 610, perform sequential density classification.Square frame 610 or the snapshot of sequential density classification step.Such as figure In 10E visible, in square frame 612, the n-th process vessel including n displacement fluid is inserted in catcher so that n More than or equal to first (i.e. second, third, the 4th etc.).It is the zoomed-in view of the cross section that VII-VII along the line obtains 1010 show the displacement fluid 312 in process vessel 302, and the cleaning fluid 1002 in Primary containers 702 and erythrocyte sedimentation rate Buffycoat 802.

Return to Fig. 6, in square frame 614, by centrifugal for system to collect fraction or subfraction, and take out the n-th processing appearance Device.In block 616, operator determines whether to obtain desired fractions or subfraction.When obtaining desired fractions or subfraction, mistake Journey can stop as shown in square frame 618, although process can continue until obtaining all fraction or subfraction.When obtaining not yet When desired fractions or subfraction, process restarts at square frame 612.Process vessel can also include process solutions, to realize Change to respective subfraction.Depend on that needs are separately and the fraction collected or subfraction number, it is possible to use two or more Plant process vessel and respective displacement fluid.Every kind in succession displacement fluid bigger than previous displacement fluid density.Similarly, Mei Gexiang Continue fraction or subfraction bigger than previous fraction or subfraction density.Once collect, it is possible to analyze continuous subfraction, such as, use In diagnosis, prognosis, research purpose, to measure component characteristics (cytometry i.e. completely), these features become the most over time Change etc..

Figure 10 F shows and experiences centrifugal catcher-process vessel system 300 and Primary containers 702.It is along the line The zoomed-in view 1012 of the viewgraph of cross-section that VIII-VIII obtains shows the stream between Primary containers 702 and process vessel 302 The snapshot of body exchange.Along with there is the cleaning fluid 1002 of the density more than buffy coat 802 in Primary containers 702 downwards Mobile, buffy coat 802 is removed from float 704.Along with having the first subfraction 1014 more than buffy coat 802 but Flow from process vessel 302 less than the displacement fluid 312 of cleaning fluid 1002 and the density of the remainder of buffy coat 802 In Primary containers 702, the first subfraction 1014 moves upwardly through funnel 222 and shelter 214 in Primary containers 702, and Enter in process vessel 302.As visible in Figure 10 G, the first subfraction 1014 is in process vessel 302, and displacement fluid 312 He Cleaning fluid 1002 is in Primary containers 702.

Process vessel 302 including the first subfraction 1014 can take out subsequently from catcher 200, with experience further Process, analyze, storage etc..After taking out process vessel 302, process solutions can be added, although process solutions can be at target Before material recovery Already in process vessel.Process vessel can such as be vibrated by turbine mixer.Exist Before vibration in liquid form, at solubilized shell or the process solutions (not shown) that adds in frangible shell, subsequently can be with Buffy coat mixes, to realize converting and forming buffy coat-process solutions mixture.Buffy coat-process solutions Mixture can be assigned on substrate such as microscope slide subsequently.

Following process container and displacement fluid may be used for collecting the other subfraction of buffy coat 802, until all Subfraction is collected or until required subfraction is collected.Although sequential density classification is described as performing with float and sealing ring, But sequential density classification can without float, sealing ring or both and perform.Hereafter it is performed for showing of sequential density classification Example method:

1. blood and float are added in pipe.

2. centrifugal with realize blood based on density separately (i.e. blood plasma, buffy coat and erythrocyte).

3. at the bottom end of float, sealing ring is applied to around pipe and float；Clamping.

4. take out blood plasma.

5. add its density having cleaning fluid more than target density.

6. inserting catcher-process vessel system, the first process vessel includes first displacement fluid with the first density.

It is centrifuged the most again.

8. taking out the first process vessel, it includes the first of the buffy coat less than the first displacement fluid density at present Subfraction.

9. being inserted in catcher by the second process vessel, described second process vessel includes that second with the second density puts Change of current body, described second density is more than the first displacement fluid and less than cleaning fluid.

It is centrifuged the most again.

11. take out the second process vessel, and it includes and ratio first displacement fluid less than the second displacement fluid density at present Second subfraction of the buffy coat bigger with the first subfraction density.

12. use the displacement fluid that density in succession is bigger to repeat step 9-11, in order to collect the substate that density in succession is bigger Point, until obtaining all required subfractions.

Target material can use any suitable analysis method or technology to be analyzed, although more specifically, and extracellular Intracellular protein labelling is included with intracellular analysis；Color staining；Analysis of molecules；Genome analysis or foranalysis of nucleic acids, including but It is not limited to gene order-checking, DNA array, expresses array, protein array and DNA hybridization array；(" ISH " uses in situ hybridization In the instrument analyzing DNA and/or RNA such as copy number changes)；Polymerase chain reaction (" PCR ")；Reverse transcription PCR；Or point Prop up DNA (" bDNA " is for analyzing the instrument of DNA and/or RNA such as mrna expression level) to analyze.These technology can need Target material before analysis is fixed, permeabilization process and separation.Some in the intracellular protein that can be labeled include But be not limited to cytokeratin (" CK "), actin, Arp2/3, coronin, dystrophin, FtsZ, myosin, Spectrin, tubulin, collagen, cathepsin D, ALDH, PBGD, Akt1, Akt2, c-myc, Caspase, survival Albumen, p27^kip, FOXC2, BRAF, phosphoric acid-Akt1 and 2, phosphoric acid-Erk1/2, Erk1/2, P38MAPK, Vimentin, ER, PgR, PI3K, pFAK, KRAS, ALKH1, Twist1, Snail1, ZEB1, fibronectin, Slug, Ki-67, M30, MAGEA3, phosphorus Acidifying receptor kinase, modified histone, chromatin-associated protein and MAGE.For fixing, permeabilization process or labelling, can make With fixative (such as formaldehyde, formalin, methanol, acetone, paraformaldehyde or glutaraldehyde), detergent (such as saponin, polyoxy second Alkene, digitonin, octyl group β-glucoside, octyl group β-glucosinolate, 1-S-octyl group β-D-thioglucopyranoside, poly-Pyrusussuriensis Alcohol ester 20, CHAPS, CHAPSO, (1,1,3,3-tetramethyl butyl) phenyl-Polyethylene Glycol or octyl phenol oxirane) or labelling Agent (such as fluorescent-labeled antibody, enzyme conjugation of antibodies, Pap stain, giemsa's stain or h and E stain).

After collection, it is also possible to make target material imaging.For imaging, the solution containing fluorescent probe may be used for labels targets Material, thus the fluorescence signal for such as being identified by imaging and characterizing is provided.Before suspension adds container, suspending After liquid adds container but before centrifugal, or after suspension has experienced and has been centrifuged, the solution containing fluorescent probe can add Enter in suspension.Fluorescent probe includes the fluorescence molecule being combined with part.Target material can have many different types of surfaces Label.Every class surface marker is the molecule that can be attached particular ligand such as antibody, such as antigen.Therefore, attached by making Being connected to part and the specific fluorescent molecular conjugate of particular surface label, part may be used for classifying target material and measuring suspension Present in the particular type of target material.The example of suitable fluorescent molecules includes but not limited to quantum dot；The dyestuff being obtained commercially, Such as fluorescein, Hoechst, FITC (" Fluorescein isothiocyanate "), R-PE (" PE "), texas Red, other algae indigo plant Albumen, Cy5, Cy7, cascade indigo plant, DAPI (" 4', 6-diamidino-2-phenylindone ") and TRITC (" tetramethylrhodamine different sulfur cyanogen Acid esters ")；Dye combinations such as CY5PE, CY7APC and CY7PE；And synthetic molecules such as self-assembly nucleic acid structure.Can make With many solution so that every kind of solution includes the different types of fluorescence molecule being combined with different ligands.

When collecting target material and mixing in non-target material, the density of target material or non-target material can increase (such as By weight (weight) being attached to target material or non-target material, or adsorb by having target material or non-target material or take in Weight), maybe can reduce (such as by buoy being attached to target material or non-target material, or by having target material or non-target Material absorption or absorption buoy).Weight or buoy can be combined with part.Target material can have many different types of surfaces Label.Every class surface marker is the molecule that can be attached particular ligand such as antibody, such as antigen.Therefore, part is permissible It is chosen as specificity and is attached to target material or non-target material.The example of approrpiate wts and/or buoy include but not limited to by metal, The pearl of glass, pottery, plastics or combinations thereof.Collecting after step and density changes step, can perform second take turns sequential Density classification, thus obtain purer target material or the various components of target material.

Aforementioned specification for illustration purposes uses specific nomenclature, to provide the thorough understanding of disclosure.But, It will be apparent to one skilled in the art that the details of concrete practice system and method described herein is optional. The described above of specific embodiments presents as an example, for the purpose of illustration and description.They are not intended to institute State thoroughly describing or making present disclosure be limited to described precise forms of precise forms.In view of instructing above, many modifications It is possible with variation.Show and describe embodiment, in order to the principle of best illustration present disclosure and practical application, from And allow those skilled in the art to optimise present disclosure and each embodiment, have as suited for the specific use considered The various modifications on way.Expection be scope of the present disclosure and is defined by appended claims and equivalent thereof.

Claims

1. reclaiming a system for target material in the sample of storage from Primary containers, described system includes:

Multiple process vessels；

There is the catcher of the first opening with relative second open fluid communication, described catcher have fit in described mainly Yardstick in the opening of container；With

Multiple displacement fluid, described multiple displacement fluid is each to be placed in different process vessels, wherein when each process vessel When inserting in described first opening, the centrifugal of described Primary containers, catcher and each process vessel promotes every kind of displacement fluid Subfraction exchange position via described catcher Yu sample fraction.

2. the system of claim 1, at least one in described multiple displacement fluid has the density more than described target material.

3. the system of claim 2, at least one in described multiple displacement fluid has the density less than described target material.

4. the system of claim 1, described multiple displacement fluid includes:

First displacement fluid, to replace the first subfraction of described sample fraction in the first process vessel；With

Second displacement fluid, so that the second subfraction of described sample fraction is replaced in the second process vessel,

Described first displacement fluid is less than described second displacement fluid density, and

Described first subfraction is less than described second subfraction density.

5. the system of claim 1, it farther includes has more than described sample, any sample fraction or any subfraction The cleaning fluid of density, at least some of sample in Primary containers described in described cleaning fluid substitution.

6. the system of claim 1, wherein said every kind in succession displacement fluid bigger than previous displacement fluid density.

7. the system of claim 1, wherein said catcher farther includes:

Including described first end and the main bodies of described opposed second ends, described main bodies includes:

Concave-shaped openings in described the second end, it narrows to the summit in described main bodies, and

There is the chamber of opening at described first end；With

Extend into intubating of intracavity from described summit, described in the opening provided between described concave-shaped openings and described chamber is provided.

8. the system of claim 1, wherein said process vessel is suction pipe head, and wherein said catcher farther includes:

Including described first end and the main bodies of described opposed second ends, described main bodies includes hole, with fluid even Connecing described first end and described opposed second ends, described hole accepts at least some of of described suction pipe head.

9. the method reclaiming target material in the sample of storage from Primary containers, described method comprises the steps:

Described Primary containers is centrifuged, with realize sample be divided into sample fraction based on density separately；

Catcher-process vessel system is introduced in the open end of described Primary containers；With

Described Primary containers is centrifuged together with described catcher-process vessel system again,

Described catcher-process vessel system includes:

Catcher, and

Including the first process vessel of first displacement fluid with the first density,

During described centrifugation step again, described first displacement fluid flow in described Primary containers via described catcher, And the first subfraction of sample fraction flow in described first process vessel via described catcher, and

Described first displacement fluid has the density more than described first subfraction.

10. the method for claim 9, wherein said first subfraction includes described target material.

The method of 11. claim 9, it farther includes following step:

Described first process vessel including described first subfraction is taken out from described catcher；

To include that the second process vessel of the second displacement fluid inserts the indoor of described catcher；With

Described Primary containers is centrifuged together with described catcher and described second process vessel again,

Described second displacement fluid flow in described Primary containers via described catcher, and the second of described sample fraction Subfraction flow in described second process vessel via described catcher,

Wherein said second displacement fluid has more than described second subfraction and the density of described first displacement fluid, and its Described in the second subfraction there is the density more than described first subfraction.

The method of 12. claim 11, wherein said first subfraction is non-target material, and described second subfraction includes institute State target material.

The method of 13. claim 11, wherein said taking-up, insertion and again centrifugation step with including the n-th of n displacement fluid Individual process vessel repeat, wherein n be more than or equal to the 3rd, and wherein said taking-up, insertion and again centrifugation step be repeated up to from Described sample obtains all required subfractions, wherein every kind in succession displacement fluid there is the close of more than every kind previous displacement fluid Degree.

The method of 14. claim 13, at least one in wherein said subfraction includes described target material, and described substate At least one in Fen does not include described target material.

The method of 15. claim 13, wherein said displacement fluid none mix with any one in other displacement fluids.

The method of 16. claim 11, wherein said first displacement fluid and the second displacement fluid do not mix.

The method of 17. claim 9, it farther includes following step:

Before centrifugation float is added in described Primary containers；With

Seal by being clamped between described float and Primary containers formation after centrifugal.

The method of 18. claim 17, it further includes at centrifugal front by the step in the cleaning fluid described Primary containers of addition Suddenly, described cleaning fluid has the density more than the most described target material.

The method of 19. claim 9, wherein said target material includes fetal material.

The method of 20. claim 19, wherein said fetal material experiences analysis of molecules after collection.

The method of 21. claim 19, in wherein said fetal material cytokeratin, alpha-Glycophorins or core label At least one be marked.

The method of 22. claim 9, it makes the step of described target material imaging after further including at collection.

The method of 23. claim 9, wherein said target material experiences analysis of molecules after collection.

The method of 24. claim 9, wherein makes at least one analyte separate with described target material.

The method of 25. claim 24, at least one analyte wherein said be trophoderm, erythroblast, fetal leukocytes, Circulating tumor cell, immunocyte or spirillum.

The method of 26. claim 9, wherein said target material include at least one spirillum, malaria derivant, immunocyte or Circulating tumor cell.

The method of 27. claim 9, wherein said catcher includes:

Including first end and the main bodies of opposed second ends, described main bodies includes:

There is the chamber of opening at described first end；With

Extend into intubating of described intracavity from described summit, described in intubate opening between described concave-shaped openings and described chamber be provided Mouthful.

The method of 28. claim 9, wherein said process vessel is suction pipe head, and wherein said catcher farther includes: