Note: Descriptions are shown in the official language in which they were submitted.
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.
13 -
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.
24
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
2123203
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.
27
~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
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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:
