CA2123203A1 - Connection-type treatment system for micro solution and method of treatment - Google Patents
Connection-type treatment system for micro solution and method of treatmentInfo
- Publication number
- CA2123203A1 CA2123203A1 CA002123203A CA2123203A CA2123203A1 CA 2123203 A1 CA2123203 A1 CA 2123203A1 CA 002123203 A CA002123203 A CA 002123203A CA 2123203 A CA2123203 A CA 2123203A CA 2123203 A1 CA2123203 A1 CA 2123203A1
- Authority
- CA
- Canada
- Prior art keywords
- container
- membrane
- connector
- open end
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
- 238000011282 treatment Methods 0.000 title claims abstract description 54
- 238000000034 method Methods 0.000 title claims abstract description 20
- 239000012528 membrane Substances 0.000 claims abstract description 73
- 238000012546 transfer Methods 0.000 claims abstract description 24
- 239000012488 sample solution Substances 0.000 claims abstract description 23
- 239000012530 fluid Substances 0.000 claims abstract description 6
- 230000002093 peripheral effect Effects 0.000 claims description 18
- 238000000108 ultra-filtration Methods 0.000 claims description 16
- 238000003780 insertion Methods 0.000 claims description 6
- 230000037431 insertion Effects 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 230000009977 dual effect Effects 0.000 claims description 3
- 230000014759 maintenance of location Effects 0.000 claims 1
- 239000000523 sample Substances 0.000 abstract description 12
- 238000005119 centrifugation Methods 0.000 abstract description 6
- 238000004458 analytical method Methods 0.000 abstract description 3
- 238000003556 assay Methods 0.000 abstract 1
- 238000006243 chemical reaction Methods 0.000 description 15
- 238000010586 diagram Methods 0.000 description 9
- 238000011109 contamination Methods 0.000 description 4
- 238000006911 enzymatic reaction Methods 0.000 description 4
- 238000004128 high performance liquid chromatography Methods 0.000 description 4
- 238000007789 sealing Methods 0.000 description 3
- 238000002372 labelling Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- -1 polypropylene Polymers 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 230000005855 radiation Effects 0.000 description 2
- 241000237074 Centris Species 0.000 description 1
- 102000004856 Lectins Human genes 0.000 description 1
- 108090001090 Lectins Proteins 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 239000004743 Polypropylene Substances 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 239000010419 fine particle Substances 0.000 description 1
- 239000003014 ion exchange membrane Substances 0.000 description 1
- 239000002523 lectin Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000005499 meniscus Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 229920001155 polypropylene Polymers 0.000 description 1
- 238000007693 zone electrophoresis Methods 0.000 description 1
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/56—Labware specially adapted for transferring fluids
- B01L3/565—Seals
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5021—Test tubes specially adapted for centrifugation purposes
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Clinical Laboratory Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Hematology (AREA)
- Separation Using Semi-Permeable Membranes (AREA)
- Sampling And Sample Adjustment (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
A connection type fluid transfer and treatment system apparatus (100) and method for efficiently and continuously executing transfer and treatment of small amounts of sample solutions without substantial transfer loss, which includes a first tube (110a) having one open end (112a) and a second closed end, a second tube (110b) having substantially the same shape as the first tube also having one open end (112b) and one closed end, and a connector assembly (126) for connecting together the open end (112a) of the first tube (110a) to the open end (112b) of the second tube (110b).
The connector assembly (126) includes a foramenous membrane support (134), which removably receives chemically or biologically treated membranes (156) for applying a predetermined treatment to a solution while passing the sample solution from the first tube (110a) to the second tube (110b). The sample is typically filtered through the membrane (156) by centrifugation. A special adaptor (170) is provided for receivingly engaging the transfer system (100) during centrifugation. The system (100) permits handling microliter quantities of reactive solutions in biochemical analyses, treatments and assays without use of micropipets, without the usual loss of solution.
The connector assembly (126) includes a foramenous membrane support (134), which removably receives chemically or biologically treated membranes (156) for applying a predetermined treatment to a solution while passing the sample solution from the first tube (110a) to the second tube (110b). The sample is typically filtered through the membrane (156) by centrifugation. A special adaptor (170) is provided for receivingly engaging the transfer system (100) during centrifugation. The system (100) permits handling microliter quantities of reactive solutions in biochemical analyses, treatments and assays without use of micropipets, without the usual loss of solution.
Description
2123~03 3/l0433-1- PCT/USg2/09789 1 8peclfiaat~0 3 Con~ otion-Typ- Tr-~t~ ~t 8y-t~
4for ~oro 801ut~0n an~ ~ t~o~ of Tr-atm ~t 6 BAC~RO~ND OF ~ INVENTIO~
7 ~ of t~- Inv ntlo~
~The present invention relat~s to a connection-type 9 transfer and treat ent syst _ and method for micro 601utions capable of perfor~ing efficient and continuous 11 transfer and/or treat~ent of a ~mall amount of sample 12 solution.
14 Br$-f D s~riptlo~ of tb- Prlor Art Conventionally, studies in the fields of analytical 16 biochemistry and clinical chemistry have been generally 17 made on the basis of working with sample treatment 18 solutions of milliliter amounts. With recent development 19 of biotechnology and immunoch~istry, however, the studies in these fields are made on the basis of results of 21 treatment of sample solutions of size on the order of 22 microlitQrs. As the treatment unit of the sample ~olution 23 becomes smaller, the following are becoming problems.
i4 In the analysis of biological samples by high performance liquid chromatography (HPLC), high performance 26 c~pillary zone electrophoresis or many other techniques, 27 pretreatment of a samples prior to analysis is often ~28 requir~d. In other cases, two or more enzymatic 29 digestions must be conducted in succes6ion to obtain the desired products. In such instances, it is necessary for 31 the sample solution, obtained by an enzyme reaction in a 32 reaction tube, to be filtered through an ultrafiltration 33 m~mhrané to remove molecules having larger molecular 34 weights or insoluble fine particles in order to prevent clogging of the high performance liquid chromatography 36 column~.
37 Typically, an instrument, such as for example a 38 micropipet, is used to transfer the sample solution from 39 the reaction tube into another device for ultrafi~tration.
W093/l~33 2 1 2 ~ 2 0 3 -2- PCT/US92/09789 1 In this method, however, a certain amount of loss of the 2 sample is inevitable in the process of transferring the 3 sample solution. The loss is greater when the sample 4 quantities are smaller.
In another example, a protein may be labeled using 6 radioisotope~, and then the labeled protein constituent 7 and the i~otopes should be ~eparated. In such cases, it -8 is conventional that, after labeling with the isotope in 9 a reaction tube, part or all of the sample solution is transferred, by micropipet or the like, into a device for 11 radiation measurement. Accordingly, the above-described 12 problem of loss of the sample also arises in the process 13 of transferring the sample solution. Also, the risk of 14 radiation contamination of instruments used in liquid transfer cannot be avoided.
16 Furthermore, when carrying out sample handling 17 procedures which by their nature require a plurality of 18 steps, such as the enzyme reaction and the sample radio-19 i~otope labeling procedures described above, the problems a~sociated with the amount of sample 108g and degree of 21 instrument contamination get progressively worse, since 22 these sample handling procedures r~quire multiple 23 transfers of the ~ample.
8~MMARY OF T~ INV~NTION
26 The invention comprises a connection type treatment 27 system and method for micro solution transfer which 28 includes: 1) a first container (source or reaction tube) 29 having a tubular shape with a first end open and an opposed second end closed, in which a reaction of a ~ample 31 solution takes place; 2) a second container (target tube) 32 of substantially the same shape as the first container 33 with oné end opened and the other end clo~ed; and 3) a 34 connector assembly for connecting the open end of the first container and the open end of the second container, 36 and also for applying predetermined treatment while 37 pa~sing the 6ample ~olution from the first container 38 (~ource tube) to the ~econd container (target tube). The 39 - connector assembly includes a connector member having a WOg3/1W33 -3- PCT/US92/09789 1 central through bore adapted to receive a membrane support 2 containing a ultra filtration membrane. A stopper fits 3 within the membrane support to hold the membrane in place.
4 In an alternate embodimQnt, the membrane support is formed integral with the connector.
6 According to another aspect of the present invention, 7 a method of treating micro solutions, using a connection 8 type treatment system for micro solutions, includes the 9 ~teps of executing a reaction of the sample solution inside the first container, connecting the open end of the 11 second container to the open end of the first container 12 u~ing the connector assembly, turning the connected first 13 and second containers upside down, and applying 14 predetermined treatment using the connector assembly while passing the sample solution from the first container into 16 the second container. At least one screw-on cap is 17 provided for sealing either or both the source and/or 18 target tubes.
19 The pre~ent invention per~it~ simultaneous transfer of a sample solution between containers as well as ~
21 predetermined treatment of the solution using two 22 containers and a specially adapted connector assembly for 23 connecting these containers. Accordingly, use of 24 transferring instruments, such as a micropipet, are not reguîred, and the problems of sample loss and 26 contamination risk are substantially reduced or minimized.
~28 BRTFF DF8CRIPTION OF TR~ DRA~G~
29 Fig. 1 is a schematic diagram of a connection-type transfer and treatment system and method for micro 31 solutions.
32 Fig. 2 is partial sectional view illustrating a 33 specif~rc structure of a centrifugal connection-type micro 34 solution transfer and treat~ent device constructed in accordance with a first embodiment of the present 36 invention.
37 Figs. 3a-3b is a ~eries of diagrams illustrating the 38 structure of the tube 10 shown in Fig. 2.
W093/1~33 2 1 2 3 2 0 3 4 PCT/USs2/09789 1 Figs. 4a-4b is a series of diagrams illustrating 2 structure of the dual tube connector 16 shown in Fig. 2.
3 Fig. 5 is a four part seriss diagram illustrating 4 structure of the filter element supporting member 22 shown in Fig. 2.
6 Figs. 6a-6b is a serie~ of diagrams illustrating 7 structure of the stopper 34 shown in Fig. 2.
-8 Fig. 7 is partial sectional view of a centrifugal 9 connection-type micro solution tran~f~r and treatment system con~tructed in accordance with a second e~bodiment 11 of the present invention.
12 Fig. 8 is a partial section view of a tube of the 13 second embodiment micro solution transfer/treatment device 14 of Fi~. 7 shown here provided with a screw-on cap 122.
Fig. 9 is a cross-sectional exploded view of the 16 second embodiment micro solution transfer/treatment device 17 of Fig. 7 shown with the upper or source tube llOb 18 omitted.
19 Fig. 10 is a top end view of the stopper lS0 of the second embodiment micro golution treatment device of Fig.
21 7 taken along the line and in the direction of arrows 10-22 10 of Fig. 9.
23 Fig. 11 is an isometric view of the stopper 150 of 24 the second embodiment device of Fig. 7.
Fig. lla is a perspective view of a tool 162 for 26 inserting the stopper 150 into the inner cylinder 130 of 27 the connector 126.
28 Fig. 12 is a top end view of the connector 126 of the 29 second embodiment micro solution treatment device illustrating the membrane support region of the connector.
31 Fig. 13 is a fragmentary cross section view of the 32 membrane support region of the connector of the second 33 embodlment micro solution treatment device taken along the 34 line and looking into the direction of arrows 13-13 of Fig. 12.
36 Fig. 14 is a side elevation view of an adapter 170 37 u~ed for securing the second embodiment for the 38 micro~olution treatment device of the present invention in 39 a centrifuge rotor.
212320~
W093/1~33 -5- PCT/US92/09789 1 Fig. 15 is a side elevation view in cross section of 2 the adap~er 170 of Fig. 14.
3 Fig. 16 is an isometric view illustrating how the 4 second embodiment micro solution treat~ent device fits within the adapter (shown in cross-section).
6 Fig. 17 i8 a functional schQmatic view in partial 7 cro~s-~ection of the second embodiment micro solution 8 treatment device of the pre~ent invention held by the 9 adapter and positioned in a fixed angle rotor.
11 D~8CRTPT~ON 0~ T9~ P~F~RR~D ~M~ODI~ENT8 12 The following detailed description illustrates the 13 invention by way of example, not by way of limitation of -14 the principles of the invention. This description will clearly enable one skilled in the art to make and use the 16 invention, and describes several embodiments, adaptations, 17 variations, alternatives and uses of the invention, 18 including what Applicant presently believes is the best 19 mode of carrying out the invention.
Fig. 1 is a diàgram which describes in schematic 21 fashion the overall system principles and method steps for 22 the connection-type micro solution transfer and treatment 23 system and method of the present invention. The presently 24 preferred embodiments of the present invention relate to a treatment system and method for pretreatment of 26 solutions for high performance liquid chromatography 27 (HPLC) using an ultrafiltration membrane.
28 Referring to Fig. 1 (a), a researcher first carries 29 out a predetermined che~ical reaction such as, for example, an enzyme reaction, in a container or tube A
31 schematically shown in Fig. 1 (a). The resulting solution 32 or product is designated by oblique lines in Fig. 1. A
33 cap (not shown) may be used on the open end of the tube.
34 Next, as is shown in Fig. 1 (b), at the end of the reaction, the experimenter then removes a cap (not shown) 36 from tube A and attaches one end of a connector C to the 37 tube A opening. A second container, indicated in the 38 drawing as container or tube B, having substantially the ~ 39 same shape as tube A, is connected in upside-down fashion WO g3/1~33 ~ 1 2 ~ 2 0 3 -6- PCT/US92/09789 1 to the other side of the connector c. The connector c 2 includes an ultrafiltration membrane (not shown) therein.
3 Next, as shown in Fig. 1 (c), the treatment 6ystem 4 integrally formed of two tubes A, B and connector C is S inverted, as shown by the intertwined arrows, and in~erted 6 in a centrifugal ~eparator D, and then the centrifugal 7 separator i8 spun. In this example, tube A i8 referred to -8 as the "source tube" or "reaction tube~ and tube B i8 9 referred to as the "target tuben.
As a result of the centrifugation, as shown in Fig.
11 1 (d), the sample solution inside reaction (source) tube 12 A passes through the ultrafiltration membrane included 13 inside connector C into the target tube B. Molecules, 14 stripped of ~olvent, having predetermined or larger molecular weights are trapped by the ultrafiltration 16 membrane.
17 As described above, according to several embodiments 18 of the present invention, the centrifugation is executed 19 with the reaction tube containing the sample solution and the tube for the centrifugation treatment being integrally 21 connected with the connector having an ultrafiltration 22 membrane therein. Therefore, by eliminating the need for 23 use of a micropipet to transfer the solution between 24 source and target tubes, there is no solution loss due to solution remaining in the micropipet instrument. Also, 26 possible contamination of the pipet is avoided. Further, 27 as compared to when solution transfer is performed by a 28 "direct pour" method whereby the contents of the reaction 29 (source) tube are poured into the target tube, virtually no sample solution residue remains on the inner source 31 tube wall in the present invention in view of the 32 completeness afforded by filtration through 33 centr~ugation.
34 When executing a reaction in a plurality of steps, the treatment in the above figures 1 (a) - (d) may be 36 repeated in each step after the second step using tube B
37 (originally the target tube), now containing the filtered 38 solution (Fig. 1 (d)), as the new reaction (source) tube 39 A', and adding a new target tube B', and so on.
21~3203 ~vo 93/10433 -7- PCI/US92/Og789 1 Fig. 2 is a partial ~ectional view of a micro 2 solution transfer/treat~ent system apparatus constructed 3 in accordance with a first embodiment of the pre~ent 4 - invention. The micro solution treatment system apparatus 1 is illustrated in a connected state corresponding to the 6 cchematic repr~sentations of Figs. 1 (c) and (d).
7 The micro solution treatment ~ystem apparatus 8 comprises reaction or source tube lOa and target tube lOb, 9 each having an open Qnd 12a, 12b oriented opposed faoing one another and ~oined together by a connector assembly 1~ 16. The tubes lOa, lOb are similarly shaped and are 12 preferably fabricated from a known plastic material of 13 the type commonly used in micro-centrifuge applications, 14 such as for example, polypropylene or polyethylene. The tubes lOa, lOb correspond to the tubes A and B of Fig. 1, 16 respectively, and the connector assembly 16 corresponds to 17 the connector C of Fig. 1.
18 The connector assembly 16 comprises a connector 19 member 17, a membrane support ~2, and a stopper 34. The connector member or connector 17 is provided with two 21 different connector ends for engagement with the tube 22 openings 12a, 12b of the respective tubes lOa, lOb 23 including a first connector end 18 defined as an open 24 mouth-type member having tapered receiving inner walls 19 dimensioned for snug, slip-fit engagement with an outer 26 peripheral wall 14a, 14b of a corresponding tube opening 27 12a or 12b, and a second connector end 20 having a male ~28 screw portion 19 provided along its outer peripheral wall 29 for engagement with a corresponding female screw portion lSa, 15b provided to an inner peripheral wall of a 31 corresponding tube opening 12a, 12b. In Fig. 2, the 32 connector 17 i8 shown having its first connector end 18 33 fitted over the outer peripheral wall 14a of tube opening 34 12a of the source tube lOa, while the male screw portion 19 of the second connector end 20 threadingly engages the 36 inner female screw portion l5b of tube opening 12b of the 37 target tube lOb.
38 The membrane support 22 is provided with a male screw 39 portion 24 formed along an outer peripheral wall and W093/1~33 2 læ 3 2 0 3 -8- PCT/US92/09789 1 having threads sized for receivingly engaging the threads 2 of the inner peripheral wall female screw portions 15a, 3 15b of a tube opening 12a, 12b. In this example, the 4 outer peripheral wall male screw portion 24 of membrane support 22 engages the inner peripheral wall female screw 6 portion 15a of the ~ource tube opening 12a. The membrane 7 support 22 i~ adjusted for receiving an ultrafiltration 8 membrane 30 placed along a bottom supporting surface 26 9 thereof (See Fig. 5). A stopper 34 i8 provided for ensuring that the membrane remains fixed within the 11 membrane support 22.
12 Fig. 3 i8 an enlarged two view diagram ~howing in 13 more detail the structure of the tube 10. In this case 14 tube 10 may be either source tube lOa or target tube lOb.
In Fig. 3, part (a) is a plan view of the tube 10 looking 16 into the tube opening 12, and part (b) is a cross-section 17 view showing the flat outer peripheral wall 14 and female 18 screw portion inner peripheral wall 15 of the tube opening 19 12. The wall thickness ~t" of the tube opening 12 preferably taper~ slightly towards its free end to permit 21 ease of insertion within the receiving connector end 18 of 22 the connector m~mber 17.
23 Fig. 4 is an enlarged two view series diagram showing 24 structure of the connector 17 of Fiq. 2 wherein part (a) is a plan view and part (b) is a cross-section view. The 26 connector 17 is generally circular in cross section and 27 includes an inner stop surface or ledge 19 against which 28 end portions of the tube opening 12 and membrane support 29 24 are constrained in abutting engagement when the system apparatus 1 is fully connected together (see Fig. 2). The 31 connector 17 is provided with a central bore hole 23 for 32 permitting transfer of solution material from a first tube 33 to a second tube connected thereto.
34 Fig. 5 is an enlarged four view series diagram illustrating the structure of the membrane support member 36 22 of Fig. 2 wherein part (a) is a top plan view 37 (supporting surface 26 omitted); part (b) is a cross 38 ~ectional view; part (c) is a side elevation view; and 39 part (d) is an enlarged bottom plan view showing the W093/1~33 -9- - PCT/US92/09789 1 configuration of a plurality of through holes or ducts 28 2 formed in the bottom wall 26 shown in part (a). Note, for 3 purpoces of clarity, the ducts 28 are not shown in the 4 cro~s ~ectional view of part (b).
Fiq. 6 is a two view series diagram illustrating 6 structure of tubular stopper 34 of Fig. 2 wherein part (a) - 7 i~ a side elevation view, and part (b) i8 a top plan view.
8 Stopper 34 resembles a ring or tubular member and includes 9 a circumferential rib 36 provided on its outer peripheral 10 wall 38 which is adapted for snap fit insertion within a 11 corresponding convex groove 27 provided to the inner 12 peripheral wall 29 of the membrane support 22 (see Fig.
13 5).
14 Combination of two tubes lOa and lOb as described 15 above can simultaneously achieve efficient transfer of 16 solutions and the centrifugation treatment as shown in 17 Fig. 1.
18 Figs. 7-13 illustrate a second embodiment for the 19 microsolution/transfer treatment system apparatus of the 20 present invention which is designated generally as element 21 100 in the drawings. Referring to ~ig. 7, the second 22 embodiment 100 for the microsolution treatment system 23 apparatus comprises two similarly shaped containers or 24 tubes llOa, llOb each having an open end 112a, 112b which 25 in use are connected together by a connector assembly 126.
26 The connector assembly 126 of the second embodiment 27 comprises two principle elements including a 28 connector/filter retainer member 127 and a stopper 150.
29 As is best seen in Figs. 7 and Fig. 9, the connector 30 member 127 is formed as a bi-annular structure having an 31 outer perimeter cylindrical shell portion or sleeve 128 32 surrounding an inner cylinder portion 130 and connected 33 integrally thereto by a lateral, radially extending web 34 132. The outer shell (sleeve) 128 and inner cylinder 35 define two connector ends including a first threaded 36 con~ector end 136 and a second slip-on connector end 140.
37 The outer shell portion or sleeve 128 is preferably 38 serrated or knurled at 137 to facilitate handling by a W093/lW33 2 1 2 ~ 2 0 ~ -lo- PCT/US92/09789 1 user. Similar grip facilitating surfaces 120a, 120b may 2 be provided to the outer ~urfaces of the tubes llOa, llOb.
3 In thi~ example, the threaded connector end 136 4 includes female screw threads disposed along an inner peripheral wall of the outer cylindrical portion 128 6 ~dapted to sngage the male screw threads 114a disposed 7 along the outer peripheral wall of the tube opening 112a 8 of tube llOa. Also, the slip on connector end 140 fits 9 over the open end 112b (and the male threads 114b) of the target tube llOb. The inner cylinder portion 130 of the 11 connector 127 al80 includes a transverse ~embrane 6upport 12 surface or region 134. In use, the connector member 127 13 i~ attached to the tube opening such that the membrane 14 supporting inner cylinder 130 is oriented to fit within the tube opening 112b of the target tube llOb. The 16 membrane support surface 134 of the inner cylinder 130 17 defines ~ foram~nous plate on which the ultrafiltration 18 membrane 156 rests. The ultrafiltration membrane 156 is 19 tightly held in place by a stopper 150 which fits within the inner cylinder 130 during use.
21 The preferred height dimension of the wall for the 22 tubular stopper 150 and inner cylinder 130 is ufficiently 23 high to ensure that all solution remains within the 24 cylindrical volume defined by the bore of tubular stopper 150 during centrifuge operation such that a meniscus, 26 which represents loss of solution, is not permitted to 27 form above the stopper 150 or cylinder 130. This volume ~28 or capacity is typically on the order of 500 ~1 to 600 ~1 29 for microsolution work. Also, the wall height of the stopper 150 is preferably slightly less than the 31 surrounding wall portion of the inner cylinder 130 so that 32 the inwardly tapered ends 158 of the stopper 150 form a 33 gradualltransition to promote full flow of fluid in the 34 downward direction from the source tube into the target tube during centrifuge operation. Also, the end walls 36 forming the mouth opening of the inner cylinder 1~0 are 37 preferably provided with a slight chamfer at 166 (see Fig~
38 9) to further promote complete flow of fluid down into the 39 inner cylinder 130.
21~3203 "'O 93/10~33 -1~ - PCI'/US92/0978g 1 Fig. 8 shows a single tube ~lo having a screw top 2 cap 122 for threading onto the outer male scrQw threads 3 114 of the tube opening 112. The cap 122 includes an 0-4 ring 124 to ensure against fluid loss. The screw on cap 122 is useful for sealing a source tube llOa, such as for 6 example after an enzyme reaction has occurred, or for 7 sealing a target tube after the desired treatment for the 8 microsolution has b~en obtained.
g Referring to Figs. 9-11, the stopper 150 includes plurality of notched relieved portions 160 spaced 11 equidistant along the top perimeter wall 154. These 12 notched portions ~60 facilitate press fit insertion of the 13 stopper within the inner cylinder membrane support 130 of 14 the connector assembly 126. The stopper 150 preferably includes a longitudinal groove (not shown) formed along 16 its outer cylindrical wall to facilitate air exchange and 17 thereby relieve any trapped air within the inner cylinder 18 membrane support 130 and the stopper 150 when the stopper 19 150 i~ fitted within the membrane inner cylinder membrane support 130. Fig. ~la illustrates an example tool 162 21 useful for inserting the stopper 150 within the inner 22 cylinder 130. The tool 162 preferably includes axially 23 extending peripheral tab members 164 for enqaging the 24 notched relieved portion 160 of the tubular stopper 150.
The top perimeter edge 154 of the ~topper 150 is 26 preferably tapered at 158 to ensure that all micro~olution 27 drains towards the ultrafiltration membrane during use and ~28 does not get trapped above the stopper perimeter edge 154.
29 Similarly, all the edges contours of the notches 160 are preferably rounded to promote and ensure fluid flow.
31 Figs. 12 and 13 illustrate in more detail the 32 generally, foramenous plate-like membrane support region 33 134 of ~he inner cylinder 130 of the connector 127. The 34 porous plate region 134 includes a plurality of arcuate and semi-arcuate througb holes or ducts 142 interspaced by 36 ribs or land portions 144. At its outer periphery the 37 membrane's support region of foramenous plate 134 includes 38 a slightly upraised rib member 146 having a peak disposed 39 coordinately aligned with lower end wall 152 of the W093/1~33 2 1 % 3 2 0 3 -12- PCT/US92/Og789 1 tubular stopper ~50 when the stopper 150 is fitted within 2 the inner cylinder 130. This is be~t seen with reference 3 to Fiq. 13 (stopper 150 and membrane ~52 ase indicated in 4 phantom). In this way, the membr~ne lS6 is maintained taut and prevented from moving by the ~ngagement of the 6 bottom end wall 152 stopper against the upraised rib 7 member 146.
8 Figs. 14-16 show an adapter 170 which may be used for g fitting the first or second embodiments of the microsolution transfer/treatment ~ystam 100 within a 11 receiving ~ocket of a centri~uge rotor. In view of the 12- added circumferential girth psovided by its additional 13 connecting elemen~s, the microsolution treatment system 14 has a slightly increased outer radiuæ as compared to conventional centrifuge tubes. Accordingly, a wider 16 diameter socket in a centrifuge rotor is preferably 17 provided for receiving the dual tube/connector system.
18 For this purpose an adapter 170 is provided to ensure 19 proper fit and support of the microsolution system 100 within the centrifuge rotor. The adapter 170 is generally 21 cylindrical`in cross section and has an inner diameter 22 sized for a close tolerance fit with the connection-type 23 microsolution system when inserted in it. The outer 24 surface of the adapter 170 is provided with a laterally extended circumferential ledge member 174 (an annular 26 flange), which acts as a stop member and rest support when 27 fitted into a receiving socket 176 of a centrifuge rotor.
28 Fig. 17 shows the system apparatus 100 placed within 29 the adapter 170 and inserted within an appropriate receiving socket or hole 176 of a rotor 178. The adapter 31 includes at its bottom end a reduced radius opening 180 32 sized to engage an outer portion of one of the tubes of 33 the microsolution system 100 at a location along the 34 bottom tube adjacent the connector assembly, such that the bottom end 182 of the system apparatus 100 is prevented 36 from contacting a base portion 184 or side wall 185 of the 37 centrifuge rotor 178. The upstanding walls 186 of the 38 adapter 170 above the ledge member 174 are of sufficient 39 length to ensure adequate support of the conne~ction-type 2123~03 ~093/1~33 -13- PCT/US92/09789 1 microsolution treatment system apparatus during centrifuge 2 operation.
3 As is best seen in Fig. 16 the forward portion of the 4 adapter may be cut away (indicated in phantom) at 188, thereby leaving only a high back supporting portion of the 6 upper adapter walls above the annular flange or ledge 7 member 174. (The cut away portion is indicated as element 8 171.) In this way, a lightweight adapter having 9 sufficient support for reducing stres~es placed on the system apparatus from centrifuged forces i8 achieved.
11 Although the above-described embodiment concerns a 12 system for pretreatment of high performance liquid 13 chromatography in which an ultrafiltration membrane is 14 provided in a connector portion, in another embodiment of the present invention a pretreatment system utilizing 16 affinity can be implemented by providing an affinity 17 functional membrane in the connection. For example, the 18 connector m~mhrane may contain antibody or antigens and 19 lectins or an ion-exchange membrane, or a membrane having other suitable functions.
21 Although the present invention has been described and 22 illustrated in detail, it should be understood that 23 various modifications within the scope of this invention 24 can be made by one of ordinary skill in the art without departing from the spirit thereof. I therefore wish my 26 invention to be defined by the scope of the appended 27 claims in view of the specification as broadly as the ~28 prior art will permit.
What is claimed is:
4for ~oro 801ut~0n an~ ~ t~o~ of Tr-atm ~t 6 BAC~RO~ND OF ~ INVENTIO~
7 ~ of t~- Inv ntlo~
~The present invention relat~s to a connection-type 9 transfer and treat ent syst _ and method for micro 601utions capable of perfor~ing efficient and continuous 11 transfer and/or treat~ent of a ~mall amount of sample 12 solution.
14 Br$-f D s~riptlo~ of tb- Prlor Art Conventionally, studies in the fields of analytical 16 biochemistry and clinical chemistry have been generally 17 made on the basis of working with sample treatment 18 solutions of milliliter amounts. With recent development 19 of biotechnology and immunoch~istry, however, the studies in these fields are made on the basis of results of 21 treatment of sample solutions of size on the order of 22 microlitQrs. As the treatment unit of the sample ~olution 23 becomes smaller, the following are becoming problems.
i4 In the analysis of biological samples by high performance liquid chromatography (HPLC), high performance 26 c~pillary zone electrophoresis or many other techniques, 27 pretreatment of a samples prior to analysis is often ~28 requir~d. In other cases, two or more enzymatic 29 digestions must be conducted in succes6ion to obtain the desired products. In such instances, it is necessary for 31 the sample solution, obtained by an enzyme reaction in a 32 reaction tube, to be filtered through an ultrafiltration 33 m~mhrané to remove molecules having larger molecular 34 weights or insoluble fine particles in order to prevent clogging of the high performance liquid chromatography 36 column~.
37 Typically, an instrument, such as for example a 38 micropipet, is used to transfer the sample solution from 39 the reaction tube into another device for ultrafi~tration.
W093/l~33 2 1 2 ~ 2 0 3 -2- PCT/US92/09789 1 In this method, however, a certain amount of loss of the 2 sample is inevitable in the process of transferring the 3 sample solution. The loss is greater when the sample 4 quantities are smaller.
In another example, a protein may be labeled using 6 radioisotope~, and then the labeled protein constituent 7 and the i~otopes should be ~eparated. In such cases, it -8 is conventional that, after labeling with the isotope in 9 a reaction tube, part or all of the sample solution is transferred, by micropipet or the like, into a device for 11 radiation measurement. Accordingly, the above-described 12 problem of loss of the sample also arises in the process 13 of transferring the sample solution. Also, the risk of 14 radiation contamination of instruments used in liquid transfer cannot be avoided.
16 Furthermore, when carrying out sample handling 17 procedures which by their nature require a plurality of 18 steps, such as the enzyme reaction and the sample radio-19 i~otope labeling procedures described above, the problems a~sociated with the amount of sample 108g and degree of 21 instrument contamination get progressively worse, since 22 these sample handling procedures r~quire multiple 23 transfers of the ~ample.
8~MMARY OF T~ INV~NTION
26 The invention comprises a connection type treatment 27 system and method for micro solution transfer which 28 includes: 1) a first container (source or reaction tube) 29 having a tubular shape with a first end open and an opposed second end closed, in which a reaction of a ~ample 31 solution takes place; 2) a second container (target tube) 32 of substantially the same shape as the first container 33 with oné end opened and the other end clo~ed; and 3) a 34 connector assembly for connecting the open end of the first container and the open end of the second container, 36 and also for applying predetermined treatment while 37 pa~sing the 6ample ~olution from the first container 38 (~ource tube) to the ~econd container (target tube). The 39 - connector assembly includes a connector member having a WOg3/1W33 -3- PCT/US92/09789 1 central through bore adapted to receive a membrane support 2 containing a ultra filtration membrane. A stopper fits 3 within the membrane support to hold the membrane in place.
4 In an alternate embodimQnt, the membrane support is formed integral with the connector.
6 According to another aspect of the present invention, 7 a method of treating micro solutions, using a connection 8 type treatment system for micro solutions, includes the 9 ~teps of executing a reaction of the sample solution inside the first container, connecting the open end of the 11 second container to the open end of the first container 12 u~ing the connector assembly, turning the connected first 13 and second containers upside down, and applying 14 predetermined treatment using the connector assembly while passing the sample solution from the first container into 16 the second container. At least one screw-on cap is 17 provided for sealing either or both the source and/or 18 target tubes.
19 The pre~ent invention per~it~ simultaneous transfer of a sample solution between containers as well as ~
21 predetermined treatment of the solution using two 22 containers and a specially adapted connector assembly for 23 connecting these containers. Accordingly, use of 24 transferring instruments, such as a micropipet, are not reguîred, and the problems of sample loss and 26 contamination risk are substantially reduced or minimized.
~28 BRTFF DF8CRIPTION OF TR~ DRA~G~
29 Fig. 1 is a schematic diagram of a connection-type transfer and treatment system and method for micro 31 solutions.
32 Fig. 2 is partial sectional view illustrating a 33 specif~rc structure of a centrifugal connection-type micro 34 solution transfer and treat~ent device constructed in accordance with a first embodiment of the present 36 invention.
37 Figs. 3a-3b is a ~eries of diagrams illustrating the 38 structure of the tube 10 shown in Fig. 2.
W093/1~33 2 1 2 3 2 0 3 4 PCT/USs2/09789 1 Figs. 4a-4b is a series of diagrams illustrating 2 structure of the dual tube connector 16 shown in Fig. 2.
3 Fig. 5 is a four part seriss diagram illustrating 4 structure of the filter element supporting member 22 shown in Fig. 2.
6 Figs. 6a-6b is a serie~ of diagrams illustrating 7 structure of the stopper 34 shown in Fig. 2.
-8 Fig. 7 is partial sectional view of a centrifugal 9 connection-type micro solution tran~f~r and treatment system con~tructed in accordance with a second e~bodiment 11 of the present invention.
12 Fig. 8 is a partial section view of a tube of the 13 second embodiment micro solution transfer/treatment device 14 of Fi~. 7 shown here provided with a screw-on cap 122.
Fig. 9 is a cross-sectional exploded view of the 16 second embodiment micro solution transfer/treatment device 17 of Fig. 7 shown with the upper or source tube llOb 18 omitted.
19 Fig. 10 is a top end view of the stopper lS0 of the second embodiment micro golution treatment device of Fig.
21 7 taken along the line and in the direction of arrows 10-22 10 of Fig. 9.
23 Fig. 11 is an isometric view of the stopper 150 of 24 the second embodiment device of Fig. 7.
Fig. lla is a perspective view of a tool 162 for 26 inserting the stopper 150 into the inner cylinder 130 of 27 the connector 126.
28 Fig. 12 is a top end view of the connector 126 of the 29 second embodiment micro solution treatment device illustrating the membrane support region of the connector.
31 Fig. 13 is a fragmentary cross section view of the 32 membrane support region of the connector of the second 33 embodlment micro solution treatment device taken along the 34 line and looking into the direction of arrows 13-13 of Fig. 12.
36 Fig. 14 is a side elevation view of an adapter 170 37 u~ed for securing the second embodiment for the 38 micro~olution treatment device of the present invention in 39 a centrifuge rotor.
212320~
W093/1~33 -5- PCT/US92/09789 1 Fig. 15 is a side elevation view in cross section of 2 the adap~er 170 of Fig. 14.
3 Fig. 16 is an isometric view illustrating how the 4 second embodiment micro solution treat~ent device fits within the adapter (shown in cross-section).
6 Fig. 17 i8 a functional schQmatic view in partial 7 cro~s-~ection of the second embodiment micro solution 8 treatment device of the pre~ent invention held by the 9 adapter and positioned in a fixed angle rotor.
11 D~8CRTPT~ON 0~ T9~ P~F~RR~D ~M~ODI~ENT8 12 The following detailed description illustrates the 13 invention by way of example, not by way of limitation of -14 the principles of the invention. This description will clearly enable one skilled in the art to make and use the 16 invention, and describes several embodiments, adaptations, 17 variations, alternatives and uses of the invention, 18 including what Applicant presently believes is the best 19 mode of carrying out the invention.
Fig. 1 is a diàgram which describes in schematic 21 fashion the overall system principles and method steps for 22 the connection-type micro solution transfer and treatment 23 system and method of the present invention. The presently 24 preferred embodiments of the present invention relate to a treatment system and method for pretreatment of 26 solutions for high performance liquid chromatography 27 (HPLC) using an ultrafiltration membrane.
28 Referring to Fig. 1 (a), a researcher first carries 29 out a predetermined che~ical reaction such as, for example, an enzyme reaction, in a container or tube A
31 schematically shown in Fig. 1 (a). The resulting solution 32 or product is designated by oblique lines in Fig. 1. A
33 cap (not shown) may be used on the open end of the tube.
34 Next, as is shown in Fig. 1 (b), at the end of the reaction, the experimenter then removes a cap (not shown) 36 from tube A and attaches one end of a connector C to the 37 tube A opening. A second container, indicated in the 38 drawing as container or tube B, having substantially the ~ 39 same shape as tube A, is connected in upside-down fashion WO g3/1~33 ~ 1 2 ~ 2 0 3 -6- PCT/US92/09789 1 to the other side of the connector c. The connector c 2 includes an ultrafiltration membrane (not shown) therein.
3 Next, as shown in Fig. 1 (c), the treatment 6ystem 4 integrally formed of two tubes A, B and connector C is S inverted, as shown by the intertwined arrows, and in~erted 6 in a centrifugal ~eparator D, and then the centrifugal 7 separator i8 spun. In this example, tube A i8 referred to -8 as the "source tube" or "reaction tube~ and tube B i8 9 referred to as the "target tuben.
As a result of the centrifugation, as shown in Fig.
11 1 (d), the sample solution inside reaction (source) tube 12 A passes through the ultrafiltration membrane included 13 inside connector C into the target tube B. Molecules, 14 stripped of ~olvent, having predetermined or larger molecular weights are trapped by the ultrafiltration 16 membrane.
17 As described above, according to several embodiments 18 of the present invention, the centrifugation is executed 19 with the reaction tube containing the sample solution and the tube for the centrifugation treatment being integrally 21 connected with the connector having an ultrafiltration 22 membrane therein. Therefore, by eliminating the need for 23 use of a micropipet to transfer the solution between 24 source and target tubes, there is no solution loss due to solution remaining in the micropipet instrument. Also, 26 possible contamination of the pipet is avoided. Further, 27 as compared to when solution transfer is performed by a 28 "direct pour" method whereby the contents of the reaction 29 (source) tube are poured into the target tube, virtually no sample solution residue remains on the inner source 31 tube wall in the present invention in view of the 32 completeness afforded by filtration through 33 centr~ugation.
34 When executing a reaction in a plurality of steps, the treatment in the above figures 1 (a) - (d) may be 36 repeated in each step after the second step using tube B
37 (originally the target tube), now containing the filtered 38 solution (Fig. 1 (d)), as the new reaction (source) tube 39 A', and adding a new target tube B', and so on.
21~3203 ~vo 93/10433 -7- PCI/US92/Og789 1 Fig. 2 is a partial ~ectional view of a micro 2 solution transfer/treat~ent system apparatus constructed 3 in accordance with a first embodiment of the pre~ent 4 - invention. The micro solution treatment system apparatus 1 is illustrated in a connected state corresponding to the 6 cchematic repr~sentations of Figs. 1 (c) and (d).
7 The micro solution treatment ~ystem apparatus 8 comprises reaction or source tube lOa and target tube lOb, 9 each having an open Qnd 12a, 12b oriented opposed faoing one another and ~oined together by a connector assembly 1~ 16. The tubes lOa, lOb are similarly shaped and are 12 preferably fabricated from a known plastic material of 13 the type commonly used in micro-centrifuge applications, 14 such as for example, polypropylene or polyethylene. The tubes lOa, lOb correspond to the tubes A and B of Fig. 1, 16 respectively, and the connector assembly 16 corresponds to 17 the connector C of Fig. 1.
18 The connector assembly 16 comprises a connector 19 member 17, a membrane support ~2, and a stopper 34. The connector member or connector 17 is provided with two 21 different connector ends for engagement with the tube 22 openings 12a, 12b of the respective tubes lOa, lOb 23 including a first connector end 18 defined as an open 24 mouth-type member having tapered receiving inner walls 19 dimensioned for snug, slip-fit engagement with an outer 26 peripheral wall 14a, 14b of a corresponding tube opening 27 12a or 12b, and a second connector end 20 having a male ~28 screw portion 19 provided along its outer peripheral wall 29 for engagement with a corresponding female screw portion lSa, 15b provided to an inner peripheral wall of a 31 corresponding tube opening 12a, 12b. In Fig. 2, the 32 connector 17 i8 shown having its first connector end 18 33 fitted over the outer peripheral wall 14a of tube opening 34 12a of the source tube lOa, while the male screw portion 19 of the second connector end 20 threadingly engages the 36 inner female screw portion l5b of tube opening 12b of the 37 target tube lOb.
38 The membrane support 22 is provided with a male screw 39 portion 24 formed along an outer peripheral wall and W093/1~33 2 læ 3 2 0 3 -8- PCT/US92/09789 1 having threads sized for receivingly engaging the threads 2 of the inner peripheral wall female screw portions 15a, 3 15b of a tube opening 12a, 12b. In this example, the 4 outer peripheral wall male screw portion 24 of membrane support 22 engages the inner peripheral wall female screw 6 portion 15a of the ~ource tube opening 12a. The membrane 7 support 22 i~ adjusted for receiving an ultrafiltration 8 membrane 30 placed along a bottom supporting surface 26 9 thereof (See Fig. 5). A stopper 34 i8 provided for ensuring that the membrane remains fixed within the 11 membrane support 22.
12 Fig. 3 i8 an enlarged two view diagram ~howing in 13 more detail the structure of the tube 10. In this case 14 tube 10 may be either source tube lOa or target tube lOb.
In Fig. 3, part (a) is a plan view of the tube 10 looking 16 into the tube opening 12, and part (b) is a cross-section 17 view showing the flat outer peripheral wall 14 and female 18 screw portion inner peripheral wall 15 of the tube opening 19 12. The wall thickness ~t" of the tube opening 12 preferably taper~ slightly towards its free end to permit 21 ease of insertion within the receiving connector end 18 of 22 the connector m~mber 17.
23 Fig. 4 is an enlarged two view series diagram showing 24 structure of the connector 17 of Fiq. 2 wherein part (a) is a plan view and part (b) is a cross-section view. The 26 connector 17 is generally circular in cross section and 27 includes an inner stop surface or ledge 19 against which 28 end portions of the tube opening 12 and membrane support 29 24 are constrained in abutting engagement when the system apparatus 1 is fully connected together (see Fig. 2). The 31 connector 17 is provided with a central bore hole 23 for 32 permitting transfer of solution material from a first tube 33 to a second tube connected thereto.
34 Fig. 5 is an enlarged four view series diagram illustrating the structure of the membrane support member 36 22 of Fig. 2 wherein part (a) is a top plan view 37 (supporting surface 26 omitted); part (b) is a cross 38 ~ectional view; part (c) is a side elevation view; and 39 part (d) is an enlarged bottom plan view showing the W093/1~33 -9- - PCT/US92/09789 1 configuration of a plurality of through holes or ducts 28 2 formed in the bottom wall 26 shown in part (a). Note, for 3 purpoces of clarity, the ducts 28 are not shown in the 4 cro~s ~ectional view of part (b).
Fiq. 6 is a two view series diagram illustrating 6 structure of tubular stopper 34 of Fig. 2 wherein part (a) - 7 i~ a side elevation view, and part (b) i8 a top plan view.
8 Stopper 34 resembles a ring or tubular member and includes 9 a circumferential rib 36 provided on its outer peripheral 10 wall 38 which is adapted for snap fit insertion within a 11 corresponding convex groove 27 provided to the inner 12 peripheral wall 29 of the membrane support 22 (see Fig.
13 5).
14 Combination of two tubes lOa and lOb as described 15 above can simultaneously achieve efficient transfer of 16 solutions and the centrifugation treatment as shown in 17 Fig. 1.
18 Figs. 7-13 illustrate a second embodiment for the 19 microsolution/transfer treatment system apparatus of the 20 present invention which is designated generally as element 21 100 in the drawings. Referring to ~ig. 7, the second 22 embodiment 100 for the microsolution treatment system 23 apparatus comprises two similarly shaped containers or 24 tubes llOa, llOb each having an open end 112a, 112b which 25 in use are connected together by a connector assembly 126.
26 The connector assembly 126 of the second embodiment 27 comprises two principle elements including a 28 connector/filter retainer member 127 and a stopper 150.
29 As is best seen in Figs. 7 and Fig. 9, the connector 30 member 127 is formed as a bi-annular structure having an 31 outer perimeter cylindrical shell portion or sleeve 128 32 surrounding an inner cylinder portion 130 and connected 33 integrally thereto by a lateral, radially extending web 34 132. The outer shell (sleeve) 128 and inner cylinder 35 define two connector ends including a first threaded 36 con~ector end 136 and a second slip-on connector end 140.
37 The outer shell portion or sleeve 128 is preferably 38 serrated or knurled at 137 to facilitate handling by a W093/lW33 2 1 2 ~ 2 0 ~ -lo- PCT/US92/09789 1 user. Similar grip facilitating surfaces 120a, 120b may 2 be provided to the outer ~urfaces of the tubes llOa, llOb.
3 In thi~ example, the threaded connector end 136 4 includes female screw threads disposed along an inner peripheral wall of the outer cylindrical portion 128 6 ~dapted to sngage the male screw threads 114a disposed 7 along the outer peripheral wall of the tube opening 112a 8 of tube llOa. Also, the slip on connector end 140 fits 9 over the open end 112b (and the male threads 114b) of the target tube llOb. The inner cylinder portion 130 of the 11 connector 127 al80 includes a transverse ~embrane 6upport 12 surface or region 134. In use, the connector member 127 13 i~ attached to the tube opening such that the membrane 14 supporting inner cylinder 130 is oriented to fit within the tube opening 112b of the target tube llOb. The 16 membrane support surface 134 of the inner cylinder 130 17 defines ~ foram~nous plate on which the ultrafiltration 18 membrane 156 rests. The ultrafiltration membrane 156 is 19 tightly held in place by a stopper 150 which fits within the inner cylinder 130 during use.
21 The preferred height dimension of the wall for the 22 tubular stopper 150 and inner cylinder 130 is ufficiently 23 high to ensure that all solution remains within the 24 cylindrical volume defined by the bore of tubular stopper 150 during centrifuge operation such that a meniscus, 26 which represents loss of solution, is not permitted to 27 form above the stopper 150 or cylinder 130. This volume ~28 or capacity is typically on the order of 500 ~1 to 600 ~1 29 for microsolution work. Also, the wall height of the stopper 150 is preferably slightly less than the 31 surrounding wall portion of the inner cylinder 130 so that 32 the inwardly tapered ends 158 of the stopper 150 form a 33 gradualltransition to promote full flow of fluid in the 34 downward direction from the source tube into the target tube during centrifuge operation. Also, the end walls 36 forming the mouth opening of the inner cylinder 1~0 are 37 preferably provided with a slight chamfer at 166 (see Fig~
38 9) to further promote complete flow of fluid down into the 39 inner cylinder 130.
21~3203 "'O 93/10~33 -1~ - PCI'/US92/0978g 1 Fig. 8 shows a single tube ~lo having a screw top 2 cap 122 for threading onto the outer male scrQw threads 3 114 of the tube opening 112. The cap 122 includes an 0-4 ring 124 to ensure against fluid loss. The screw on cap 122 is useful for sealing a source tube llOa, such as for 6 example after an enzyme reaction has occurred, or for 7 sealing a target tube after the desired treatment for the 8 microsolution has b~en obtained.
g Referring to Figs. 9-11, the stopper 150 includes plurality of notched relieved portions 160 spaced 11 equidistant along the top perimeter wall 154. These 12 notched portions ~60 facilitate press fit insertion of the 13 stopper within the inner cylinder membrane support 130 of 14 the connector assembly 126. The stopper 150 preferably includes a longitudinal groove (not shown) formed along 16 its outer cylindrical wall to facilitate air exchange and 17 thereby relieve any trapped air within the inner cylinder 18 membrane support 130 and the stopper 150 when the stopper 19 150 i~ fitted within the membrane inner cylinder membrane support 130. Fig. ~la illustrates an example tool 162 21 useful for inserting the stopper 150 within the inner 22 cylinder 130. The tool 162 preferably includes axially 23 extending peripheral tab members 164 for enqaging the 24 notched relieved portion 160 of the tubular stopper 150.
The top perimeter edge 154 of the ~topper 150 is 26 preferably tapered at 158 to ensure that all micro~olution 27 drains towards the ultrafiltration membrane during use and ~28 does not get trapped above the stopper perimeter edge 154.
29 Similarly, all the edges contours of the notches 160 are preferably rounded to promote and ensure fluid flow.
31 Figs. 12 and 13 illustrate in more detail the 32 generally, foramenous plate-like membrane support region 33 134 of ~he inner cylinder 130 of the connector 127. The 34 porous plate region 134 includes a plurality of arcuate and semi-arcuate througb holes or ducts 142 interspaced by 36 ribs or land portions 144. At its outer periphery the 37 membrane's support region of foramenous plate 134 includes 38 a slightly upraised rib member 146 having a peak disposed 39 coordinately aligned with lower end wall 152 of the W093/1~33 2 1 % 3 2 0 3 -12- PCT/US92/Og789 1 tubular stopper ~50 when the stopper 150 is fitted within 2 the inner cylinder 130. This is be~t seen with reference 3 to Fiq. 13 (stopper 150 and membrane ~52 ase indicated in 4 phantom). In this way, the membr~ne lS6 is maintained taut and prevented from moving by the ~ngagement of the 6 bottom end wall 152 stopper against the upraised rib 7 member 146.
8 Figs. 14-16 show an adapter 170 which may be used for g fitting the first or second embodiments of the microsolution transfer/treatment ~ystam 100 within a 11 receiving ~ocket of a centri~uge rotor. In view of the 12- added circumferential girth psovided by its additional 13 connecting elemen~s, the microsolution treatment system 14 has a slightly increased outer radiuæ as compared to conventional centrifuge tubes. Accordingly, a wider 16 diameter socket in a centrifuge rotor is preferably 17 provided for receiving the dual tube/connector system.
18 For this purpose an adapter 170 is provided to ensure 19 proper fit and support of the microsolution system 100 within the centrifuge rotor. The adapter 170 is generally 21 cylindrical`in cross section and has an inner diameter 22 sized for a close tolerance fit with the connection-type 23 microsolution system when inserted in it. The outer 24 surface of the adapter 170 is provided with a laterally extended circumferential ledge member 174 (an annular 26 flange), which acts as a stop member and rest support when 27 fitted into a receiving socket 176 of a centrifuge rotor.
28 Fig. 17 shows the system apparatus 100 placed within 29 the adapter 170 and inserted within an appropriate receiving socket or hole 176 of a rotor 178. The adapter 31 includes at its bottom end a reduced radius opening 180 32 sized to engage an outer portion of one of the tubes of 33 the microsolution system 100 at a location along the 34 bottom tube adjacent the connector assembly, such that the bottom end 182 of the system apparatus 100 is prevented 36 from contacting a base portion 184 or side wall 185 of the 37 centrifuge rotor 178. The upstanding walls 186 of the 38 adapter 170 above the ledge member 174 are of sufficient 39 length to ensure adequate support of the conne~ction-type 2123~03 ~093/1~33 -13- PCT/US92/09789 1 microsolution treatment system apparatus during centrifuge 2 operation.
3 As is best seen in Fig. 16 the forward portion of the 4 adapter may be cut away (indicated in phantom) at 188, thereby leaving only a high back supporting portion of the 6 upper adapter walls above the annular flange or ledge 7 member 174. (The cut away portion is indicated as element 8 171.) In this way, a lightweight adapter having 9 sufficient support for reducing stres~es placed on the system apparatus from centrifuged forces i8 achieved.
11 Although the above-described embodiment concerns a 12 system for pretreatment of high performance liquid 13 chromatography in which an ultrafiltration membrane is 14 provided in a connector portion, in another embodiment of the present invention a pretreatment system utilizing 16 affinity can be implemented by providing an affinity 17 functional membrane in the connection. For example, the 18 connector m~mhrane may contain antibody or antigens and 19 lectins or an ion-exchange membrane, or a membrane having other suitable functions.
21 Although the present invention has been described and 22 illustrated in detail, it should be understood that 23 various modifications within the scope of this invention 24 can be made by one of ordinary skill in the art without departing from the spirit thereof. I therefore wish my 26 invention to be defined by the scope of the appended 27 claims in view of the specification as broadly as the ~28 prior art will permit.
What is claimed is:
Claims (11)
1. A connection-type transfer and treatment system for micro solutions comprising in operative combination:
a) a first container of a generally tubular shape having a first open end and a second closed end, said first container adapted to contain a sample solution therein;
b) a second container having substantially the same shape as said first container and having a first open end and a second closed end;
c) connector means for connecting the open end of said first container to the open end of said second container; and d) means for retaining a membrane for filtering or/and applying a predetermined treatment to said sample solution while passing said sample solution from said first container into said second container, said membrane retaining means disposed internal of said connecting means to contact fluid passing from said first to said second container.
a) a first container of a generally tubular shape having a first open end and a second closed end, said first container adapted to contain a sample solution therein;
b) a second container having substantially the same shape as said first container and having a first open end and a second closed end;
c) connector means for connecting the open end of said first container to the open end of said second container; and d) means for retaining a membrane for filtering or/and applying a predetermined treatment to said sample solution while passing said sample solution from said first container into said second container, said membrane retaining means disposed internal of said connecting means to contact fluid passing from said first to said second container.
2. A connection-type treatment system for micro solutions as in claim 1 wherein:
a) said connecting means is reversibly connectable for exchanging the position of said first and second containers to permit successive treatments to the micro solution while passing between said first and second containers.
a) said connecting means is reversibly connectable for exchanging the position of said first and second containers to permit successive treatments to the micro solution while passing between said first and second containers.
3. A connection-type treatment system for micro solutions as in claim 2 wherein:
a) each of said containers includes threads disposed along a peripheral wall surface adjacent its open end; and b) said connecting means is a connector assembly for mechanically connecting said first and second containers and wherein said connector assembly includes:
i) a sleeve member having a first connector end for slip fit connection with said open end of either said first or second containers, and a second end including threads for engagement with said threads of the other one of said first or second container open ends;
ii) a membrane support adapted for fitted insertion within said sleeve connector member adjacent said first connector end and having a central base for passing micro solution therethrough and a generally flat foramenous surface for retaining said membrane means; and iii) means for securing said membrane means to said membrane support in a manner to provide a liquid tight seal between the periphery of said membrane support and said membrane means.
a) each of said containers includes threads disposed along a peripheral wall surface adjacent its open end; and b) said connecting means is a connector assembly for mechanically connecting said first and second containers and wherein said connector assembly includes:
i) a sleeve member having a first connector end for slip fit connection with said open end of either said first or second containers, and a second end including threads for engagement with said threads of the other one of said first or second container open ends;
ii) a membrane support adapted for fitted insertion within said sleeve connector member adjacent said first connector end and having a central base for passing micro solution therethrough and a generally flat foramenous surface for retaining said membrane means; and iii) means for securing said membrane means to said membrane support in a manner to provide a liquid tight seal between the periphery of said membrane support and said membrane means.
4. A connection-type treatment system for micro solutions as in claim 3 wherein said membrane support includes threads along an outer peripheral wall thereof for threaded engagement with an unoccupied one of said screw means of either said first or second container located on the interior wall of said containers adjacent the open ends thereof.
5. A connection-type treatment system for micro solutions as in claim 4 wherein said membrane means is an ultrafiltration membrane and said securing means for said membrane means is a stopper member in the shape of a ring having an outer surface configuration adapted for snap fit insertion within said central cavity of said membrane support.
6. A connection-type treatment system for micro solutions as in claim 2 wherein;
a) each of said containers includes threads disposed along a peripheral wall surface adjacent its open end;
b) said connecting means is a connector assembly for mechanically connecting together said first and second containers wherein said connector assembly includes a connector member including an outer sleeve portion and integrally attached inner tubular membrane support, said connector member having a first connector end for slip fit connection with said open end of either said first or second containers and a second connector end including threads for engagement of said threads of the other one of said first or second container open ends; said membrane support includes a generally flat foramenous surface for receiving said membrane means; and means for securing said membrane means to said membrane support in a manner to provide a liquid tight seal between a periphery of said membrane support and said membrane means and to minimize retention of fluid in said connector and said first container.
a) each of said containers includes threads disposed along a peripheral wall surface adjacent its open end;
b) said connecting means is a connector assembly for mechanically connecting together said first and second containers wherein said connector assembly includes a connector member including an outer sleeve portion and integrally attached inner tubular membrane support, said connector member having a first connector end for slip fit connection with said open end of either said first or second containers and a second connector end including threads for engagement of said threads of the other one of said first or second container open ends; said membrane support includes a generally flat foramenous surface for receiving said membrane means; and means for securing said membrane means to said membrane support in a manner to provide a liquid tight seal between a periphery of said membrane support and said membrane means and to minimize retention of fluid in said connector and said first container.
7. A connection-type treatment system for micro solutions as in claim 6 wherein:
a) said securing means for said membrane means includes a generally tubular stopper member adapted for fitted insertion within said membrane support and having a bottom end wall co-aligned with an upraised perimeter rib member provided to said foramenous surface of said membrane support for pinning said membrane means to said membrane support.
a) said securing means for said membrane means includes a generally tubular stopper member adapted for fitted insertion within said membrane support and having a bottom end wall co-aligned with an upraised perimeter rib member provided to said foramenous surface of said membrane support for pinning said membrane means to said membrane support.
8. A connection-type treatment system for micro solutions as in claim 7 which includes an ultrafiltration membrane and wherein said outer surface portions of said first and second containers and said outer sleeve of said connector member are knurled to facilitate handling by a user.
9. A connection-type treatment system for micro solutions comprising in operative combination:
a) a first container of a generally tubular shape having a first open end and a second closed end, said first container is adapted to retain a sample solution therein;
b) a second container having a substantially similar shape as said first container and having a first open end and a second closed end;
c) connector means for connecting the open end of said first container to the open end of said second container;
d) means for retaining a membrane for filtering and/or applying a predetermined treatment to said sample solution while passing said sample solution from said first container into said second container, said membrane means disposed internal of said connecting means;
e) a tubular adapter receivingly engaging at least said connector for properly positioning and supporting said dual container and connector assembly within a centrifuge rotor when said first and second containers are connected by said connector means; and f) an ultrafiltration membrane including a component for treating said solution during passage therethrough.
a) a first container of a generally tubular shape having a first open end and a second closed end, said first container is adapted to retain a sample solution therein;
b) a second container having a substantially similar shape as said first container and having a first open end and a second closed end;
c) connector means for connecting the open end of said first container to the open end of said second container;
d) means for retaining a membrane for filtering and/or applying a predetermined treatment to said sample solution while passing said sample solution from said first container into said second container, said membrane means disposed internal of said connecting means;
e) a tubular adapter receivingly engaging at least said connector for properly positioning and supporting said dual container and connector assembly within a centrifuge rotor when said first and second containers are connected by said connector means; and f) an ultrafiltration membrane including a component for treating said solution during passage therethrough.
10. A method of transferring and treating micro solutions using a connection type micro solution treatment system which includes a pair of tubular containers of substantially similar shape, each container having one end opened and the other end closed, connector means for connecting the open ends of said containers together, and membrane means carried by said connector means for filtering and/or applying predetermined treatment to the micro solution, said method comprising the steps of:
a) introducing a sample solution into a first of said container pair;
b) emplacing an ultrafiltration membrane having a solution treatment component associated therewith in said connector;
c) connecting the open end of a second container to the open end of a first container using said connector means with said first container being below said second container;
d) inverting said connected pair of containers so that said sample solution can pass through said membrane into said second container therebelow; and d) treating said solution while passing the sample solution from said first container into said second container.
a) introducing a sample solution into a first of said container pair;
b) emplacing an ultrafiltration membrane having a solution treatment component associated therewith in said connector;
c) connecting the open end of a second container to the open end of a first container using said connector means with said first container being below said second container;
d) inverting said connected pair of containers so that said sample solution can pass through said membrane into said second container therebelow; and d) treating said solution while passing the sample solution from said first container into said second container.
11. The treatment method as in claim 10 wherein said step of treating includes the steps of:
a) placing said connection type treatment system for micro solution within a centrifuge rotor; and b) centrifuging until said micro solution from said first container passes through said membrane means and is collected in said second container.
a) placing said connection type treatment system for micro solution within a centrifuge rotor; and b) centrifuging until said micro solution from said first container passes through said membrane means and is collected in said second container.
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US79183791A | 1991-11-14 | 1991-11-14 | |
| US07/791,837 | 1991-11-14 | ||
| US93001792A | 1992-08-13 | 1992-08-13 | |
| US07/930,017 | 1992-08-13 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2123203A1 true CA2123203A1 (en) | 1993-05-27 |
Family
ID=27121206
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002123203A Abandoned CA2123203A1 (en) | 1991-11-14 | 1992-11-13 | Connection-type treatment system for micro solution and method of treatment |
Country Status (7)
| Country | Link |
|---|---|
| EP (1) | EP0612403B1 (en) |
| JP (1) | JPH07501150A (en) |
| AU (1) | AU3135093A (en) |
| CA (1) | CA2123203A1 (en) |
| DE (1) | DE69228386T2 (en) |
| TW (1) | TW209853B (en) |
| WO (1) | WO1993010433A1 (en) |
Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB9315779D0 (en) * | 1993-07-30 | 1993-09-15 | Stoneplan Limited | Apparatus and methods |
| AU702874B2 (en) * | 1993-07-30 | 1999-03-11 | Kidde Products Limited | Smoke detection system |
| DE9418060U1 (en) * | 1994-11-11 | 1996-03-14 | SC - Sanguis Counting Kontrollblutherstellungs- und Vertriebs GmbH, 51588 Nümbrecht | Sample tube and end cap, especially for capillary blood collection |
| US5603900A (en) * | 1995-05-19 | 1997-02-18 | Millipore Investment Holdings Limited | Vacuum filter device |
| GB2321857B (en) * | 1997-02-05 | 2000-05-24 | Intersep Ltd | Improvements in filters |
| DE10141817B4 (en) | 2001-08-27 | 2005-03-03 | Eppendorf Ag | Membrane device for receiving samples and method for producing a device |
| WO2003019131A2 (en) | 2001-08-29 | 2003-03-06 | Hexal Pharma Gmbh | Method and device for preparing a sample of biological origin in order to determine at least one constituent contained therein |
| EP2768594B1 (en) * | 2011-10-18 | 2023-06-07 | The Trustees of Columbia University in the City of New York | Medical apparatus and method for collecting biological samples |
| EP3489651B1 (en) | 2012-11-20 | 2021-08-04 | The Trustees of Columbia University in the City of New York | Medical apparatus for collecting biological samples |
| PL3433527T3 (en) * | 2016-03-23 | 2022-03-21 | Husqvarna Ab | Tap connector with flow stabilization |
| CN110146583B (en) * | 2019-04-26 | 2020-08-11 | 中国科学院地质与地球物理研究所 | Method for decomposing sample by Carius tube and method for analyzing isotope of Re-Os |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3701434A (en) * | 1971-03-15 | 1972-10-31 | Hugh C Moore | Test tube system for separating blood into serum and red cells |
| US3802843A (en) * | 1971-05-28 | 1974-04-09 | American Hospital Supply Corp | Fluid testing apparatus |
| US4632761A (en) * | 1983-08-15 | 1986-12-30 | W. R. Grace & Co. | Centrifugal microconcentrator and methods for its use |
| US4678559A (en) * | 1984-07-23 | 1987-07-07 | Andreas Szabados | Test specimen container for pasty specimen material |
| US4675110A (en) * | 1986-01-31 | 1987-06-23 | Biomedical Polymers, Inc. | Filter device and apparatus with multiple gas return passages |
| US4832678A (en) * | 1987-12-03 | 1989-05-23 | E. I. Du Pont De Nemours And Company | Adapter for a centrifuge tube and a removal tool therefor |
-
1992
- 1992-11-13 DE DE69228386T patent/DE69228386T2/en not_active Expired - Fee Related
- 1992-11-13 JP JP50941393A patent/JPH07501150A/en active Pending
- 1992-11-13 AU AU31350/93A patent/AU3135093A/en not_active Abandoned
- 1992-11-13 CA CA002123203A patent/CA2123203A1/en not_active Abandoned
- 1992-11-13 WO PCT/US1992/009789 patent/WO1993010433A1/en active IP Right Grant
- 1992-11-13 EP EP92925198A patent/EP0612403B1/en not_active Expired - Lifetime
- 1992-11-20 TW TW81109320A patent/TW209853B/zh active
Also Published As
| Publication number | Publication date |
|---|---|
| WO1993010433A1 (en) | 1993-05-27 |
| DE69228386T2 (en) | 1999-09-30 |
| JPH07501150A (en) | 1995-02-02 |
| DE69228386D1 (en) | 1999-03-18 |
| EP0612403A4 (en) | 1994-12-28 |
| AU3135093A (en) | 1993-06-15 |
| EP0612403B1 (en) | 1999-02-03 |
| TW209853B (en) | 1993-07-21 |
| EP0612403A1 (en) | 1994-08-31 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| FZDE | Discontinued | ||
| FZDE | Discontinued |
Effective date: 20001114 |