US 3096283 A
Description (OCR text may contain errors)
July 2., 1963 G. N. HEIN 3,096,283
CONTAINER FOR BLOOD AND MACHINE FOR SEPARATING PRECIPITATES FROM LIQUID BLOOD CONSTITUENTS Filed June 24, 1959 4 Sheets-Sheet 1 INVEN TOR. GEO/P65 M ////1/ Bax/ 04,444,
July 2, 1963 c; N HElN 3 096,283
9 CONTAINER FOR BLOOD AND MACHINE FOR SEPARATING PRECIPITATES FROM LIQUID BLOOD CONSTITUENTS Filed June 24, 1959 4 Sheets-Sheet 2 FIG. 2
INVENTOR. f 650%,; A/- AE/A/ BY a D July 2,, 1963 G. N. CONTAINER FOR BLOOD AND HEIN MACHINE FOR SEPARATI PRECIPITATES FROM LIQUID BLOOD CONSTITUENTS Filed June 24, 1959 4 Sheets-Sheet 3 July 2., 1963 G. N. HEIN 3,096,283
CONTAINER FOR BLOOD AND MACHINE FOR SEPARATING PRECIPITATES FROM LIQUID BLOOD CONSTITUENTS Filed June 24, 1959 4 Sheets-Sheet 4 FIG. 5
IN V EN T 0R. GEOPE /V- #E/A/ United States Patent Oiiice ,i 2,
3,tl96,283 CONTAINER FQR BLQOD AND MACI-IENE lifiR SEPARATBNG PRECEPITATES FROM LIQUED BLOOD CONSTITUENTS George N. Hein, San Carlos, Calif, assignor to Becton,
Dickinson and (Iompany, Rutherford, Ni, 21 corporation of New Jersey Filed June 24, 1959, Ser. No. 822,561 Claims; (Cl.- 2332tl) This invention relates to a functionally and structurally improved receptacle and machine which, when properly used in association with each other, will substantially completely separate precipitates in blood from the liquid plasma or serum.
It is an object of the invention to teach a novel machine and receptacle for use therewith for separating whole blood into its constituents in a manner such that the remaining liquid (whether plasma or serum) will be clear and in effect free from solids.
A further object is that of teaching an apparatus and receptacle for use therewith whereby these solids will be in turn subdivided in a manner such that white cells, red cells, platelets, fats, etc. may also be separated or stratified with respect to each other.
Still another object is that of furnishing a receptacle suitable for the reception, storage and treatment of blood, and in which, after separation, the clear liquid serum and plasma may readily be used; the solid constituents being subject to ready. examination and analysis, where such a step is to be resorted to. 7 An additional object is that of furnishing a machine of relatively simple design and with which the receptacle may readily be associated in accomplishing such separation.
With these and other objects in mind, reference is had to the attached sheets of drawings illustrating practical embodiments of the invention, and in which:
FIG. 1 is a fragmentary side elevation of one form of the apparatus as well as the receptacle for use in connection with the same, the parts being separated so that their relationship may more readily be visualized;
FIG. 2 is a sectionalside view showing the parts in assembled relationship and in a condition of rest;
FIG. 3 is a view similar to FIG. 2, illustrating the parts of the assembly in diiferent positions and also showing the mechanism in a condition of rest;
FIG. 4 is a sectional side view of one improved form of container for the blood;
FIG. 5 is a sectional side view of an alternative form of operating head to thatillustrated in FIGS. 1, 2 and v3;
FIG. 6 is a similar view of afurther form of operating head; and
FIG. 7 shows an assembly for mounting a receptacle of the present type and by'rneans of which that receptacle maybe collapsed or expanded.
Referring primarily to FIG. 1, the numeral 10 indicates a ring-shaped element forming a part of the machine and which overlies a bowl 11 to be secured against movement with respect thereto by inter-engagingscrew threads 12 formed respectively on the outer face of the bowl and the inner face of ring 10. Below bowl 11 an actuator ring 13 is disposed and has extending from it asuitable number 'of handles 14. This ring is internally threaded and mounted upon the thread 16 of a cup member, in turn supported above a motor casing 19. Extending through the latter is'a hollow tube 20. Conveniently below the motor casing, avtubular extension 21 is provided. Into the latterthere extends a screw-threaded stem 22 terminating in an actuating portion 23.
. As'also shown in this viewand FIG. 2, a blood receptaclesuitable for association with the machine is present.
That receptacle is conveniently formed of clear plastic, such as vinyl. As illustrated, it will include a pan-shaped base portion 24, above which there is disposed an upper portion 25. That upper portion may in effect include an inverted pan-shaped structure, but in any'event, it is provided with a central opening conveniently defined by an upstanding flange 26, closed by a headed stopper 27, preferably of rubber, having perforable diaphragm portions 23 which may be penetrated by a cannula. The base part 24 and upper part 25 of the receptacle are coupled by an annulus '28, preferably integral therewith. In many instances it will be preferred to make the upper part of a material, or of sufficient gauge, so that it will be relatively rigid. Both the base 24 and part 28 may be relatively flexible.
In FIG. 2, in which the interior of the assembly has been illustrated, it will be understood that as shown, the flexible plastic receptacle may be provided with a cap 2? of aluminum or other suitable material, which will serve to retain stopper 27 against displacement from the neck portion 26 of the receptacle. The ring it? has its underface channeled, as indicated at 39, so as to receive part of the rim portion or annulus 23 of the receptacle. Similarly, the upper inner edge zone of bowl 11 may be recessed, as at 31, to accommodate part of this receptacle portion. Ring ltl is downwardly and inwardly inclined so as to provide at 32 a clamp ledge or zone. A cooperating clamping zone 33 forms a part of a supporting and manipulating member 34 within bowl 11. interposed between the supporting and clamping member 34 and the inner face of the bowl is a bezel 35 provided with an inwardly extending portion 35. The upper edge of this bezel, which functions as an expressing member, is in line with the lower central zone of annulus 28 when the receptacle is in position. The pins 18 extend through packings such as O-rings in the base of bowl 11 and engage the inwardly extending bezel portion 26. If desired, the upper portion 25 of the receptacle may be overlain by a support 37 of frustroconical shape and of any desired material, such as metal.
Secured to bowl 11 by, for example, a screw-threaded connection is a block 3% which provides in its upperv face a cylinder slidably supporting a piston 4d. The skirt of the latter may slidably engage a centrally disposed projecting portion 41 integral with or extending from the support and clamp 34. The tube 20 may either provide the shaft for the motor disposed within casing 19, (FIG. 1), or else be connected so that it will rotate in unison with that shaft. This tube has a bore 42 extending throughout its length. Adjacent the lower end of the tube, the bore is conveniently enlarged to furnish a cylinder within which a piston or plunger 43 is slidably disposed. That plunger is projectable and retractable by securing the inner end of the stem 22 to a cup 44, which supports a bearing 45 carrying a plate 46. The lower end of the plunger is coupled with that plate. Therefore, as stem 22 is projected and retracted, the plunger will be correspondingly moved.
Any suitable number of bearings of the type indicated at 47 may encircle tube or shaft 29 to rotatably support the latter. Also, a thrust bearing or bearings (not shown) may be provided. A further bearing 43 may have one of its raceways secured to the actuating ring 13'. Its opposite raceway bears against plate 15. It is apparent that as the ring or collar is rotated, and due to the engagement of its bore threads with thread 16 of cup 17, that collar will be elevated or lowered, according to the directon of rotation. As it elevates, it will cause pins 18 to bear against flange 36 of bezel ring 35 to project the latter.
Access may be gained to the interior of annulus 28 even while the latter is in association with the machine.
If this result is desired, ring may be provided with one or more openings in its upper face. These openings are sealed by stoppers 4 of the puncture type and which are preferably formed of rubber. The stoppers will overlie the precipitate chamber provided by the annulus. Therefore, a cannula in the form of a hollow needle, as at 50 (FIG. 3), may be caused to puncture a stopper 4-9, so that the inner end of the needle enters the interior of the ring portion. Obviously, as the needle is withdrawn, the stopper will reestablish the seal. The parts may be so constructed that renewal of the stopper or stoppers may be effected when necessary.
A preferred procedure would involve dismounting ring 10 from bowl 11 and disposing in proper position within the latter a receptacle such as 24-25. This receptacle either would contain whole blood, or else might receive it after the reapplication of ring 19 to bowl 11, by, for example, employing hollow needles for filling and venting purposes and forcing the points of those needles through the properdiaphragm portion 28. If before processing, the blood was mixed with an anticoagulant solution, the resultant material, after completion of the operation, would be grouped on the one hand in the form of a clear plasma filtrate liquid, and on the other hand in the form of solids. The latter would consist of white cells, red cells, platelets, etc. If the blood had not been mixed with anti-coagulant prior to the practice of the present method, the resultant liquidafter processingwould be a clear serum filtrate.
With the ring applied to the bowl, the receptacle will be properly supported by the mounting and clamp member 34. It will be retained against turning with respect to the rotor assembly by having its periphery clamped between the element 10 and bowl member 11. Its precipitate chamber, as defined by the annulus 28, will be housed and supported by the recessed surfaces incorporated in ring 10, the peripheral portion of bowl 11 and the adjacent portions of members 35 and 34.
In any event, with the parts properly disposed and plunger 43 and pins 18 retracted, the precipitate chamber 23 will be in fully distended position. Also, the inner edge zones of that chamber, which continue in the form of parts 24 and 25, will be spaced in lines with the clamping portions 32 and 33. Such spacing may be on the order of .020". The motor which serves to rotate shaft or tube 20 will now be energized. As it operates, it may cause a turning of that shaft at a speed of around 10,000 rpm. With centrifugal force thus created, the solids or other components of high specific gravity within the whole blood will be thrown outwardly and will pass into the annulus 28. This will leave within the main chamber of the receptacle, as defined by parts 24 and 25, a clear liquid free from solids. While the parts continue to rotate at high speed, any excess liquid unnecessarily deposited within the precipitate chamber may be returned to the main body of the receptacle. This is achieved by grasping handles 14 and turning the actuating ring 13. With such turning in a proper direction, that ring will shift toward bowl 1-1. So shifted, pins 18 will engage flange 36 of bezel 35 to move the latter from the position shown in FIG. 2 to that illustrated in FIG. 3. This action may be continued until substantially all serum or plasma has been displaced inwardly from chamber 28 through the annular space existing between clamping portions 32 and 33.
Now, by actuating stem 22, plunger 43 will be projected. As shown, O-rings are provided in association with the liquid control system of the apparatus. Therefore, with liquid, such as oil within tube 20, it will be forced up through the cylinder within which piston 45 rides, and so through bore 42 into the cylinder formed in block 39. Within the latter, it will serve to shift piston 40 toward the receptacle 2425. So shifting, it will move from the position shown in FIG. 2 to that shown in FIG. 3. Under these circumstances, its skirt will engage the underface of the supporting and clamping member 34 and elevate the latter to thus close the gap between sealing or clamping portions 32-33. Therefore, the chamber defined within annulus 28 will be completely sealed from the main body of the receptacle, and the solids within that chamber will be isolated from the liquid. If at this time it is desired to withdraw samples of the solids, this may be done without removing the container from the machine, by simply causing a hollow needle to perforate the stopper or stoppers 4?. Of course, prior to such procedure, rotation of the parts will have been interrupted.
It is preferred that the motor within casing 19 be subject to control so that it can rotate at relatively slow speeds. A preferred method of procedure involves such a rotation with a receptacle in position within the ring and bowl. That receptacle would not be provided with a sealing cap, and in fact might not include a pouring neck or flange 26. Rather, the upper end of the receptacle would simply present an opening through which oxalated blood could be poured or passed into the interior of the receptacle. Now the speed of rotation is gradually increased up to around 10,000 rpm, with which a centrifugal force of not less than two thousand times gravity may be developed. This will minimize hemolysis. The plasma will quickly become clear. After sealing of the precipitate chamber has been effected, then that seal should be maintained until the plasma or serum has been removed from the container.
Thus, it will be understood that the present invention offers a radical improvement over procedures as heretofore practiced. One such procedure involves the placing of the whole blood in a bottle or other receptacle and permitting it to stand for three or four hours. Under these circumstances, the majority of the solids will settle to the bottom of the receptacle. Such a procedure does not, of course, compact the cells, and produces a plasma or sen-um which will still retain many solids in suspension. Also, the plasma or serum must be Withdrawn without disturbing the solids which have settled to the bottom.
With the other heretofore accepted method, suitable receptacles are used to receive the blood, and rotation is effected at a speed of approximately 2000 r.p.m. Under these conditions, the precipitates are forced to the bottom of the vessel. This force, however, is. not sufficient to remove all solids from the plasma or serum. This is especially true in that when the centrifuge is once stopped, some of the solids will again mix with the filtrate. At best, the problem of removing that filtrate is a diflicult one, and solids again remain in suspension within it.
One preferred form of container has been generally illustrated in FIG. 4. In that view it will be seen this unit involves an upper section 51, preferably in the form of a truncated cone, and having a central opening 52 through which liquids may be introduced or withdrawn. A base section 53 corresponds to section 51, except that it is imperforate. Both sections terminate in peripheral portions providing in aggregate an annulus 54, and may have their outer edge zones sealed in face-to-face contact, as indicated at 55. It is apparent that the receptacle might take various other alternative forms, but the two foregoing examples, in addition to those of FIGS. 5 and 6, will be adequate for the purpose of visualizing the range of these forms.
A bowl assembly as shown in FIG. 5 may be resorted to, in which a ring 51' is provided corresponding to the ring 10 heretofore described. Also, a bowl 52', similar to bowl 11, has screw-threaded connection therewith. Within this bowl, a support and clamping member 53 is disposed. The base of the bowl conveniently includes an extension 54 formed with sockets 55'. Also, set screws 56 are carried by the bowl, and when projected, bear against the underface of supporting member 53'. Thus, after the parts have been rotated and the Solids 5. separated'fromthe liquid, the clamping member 53' may be shifted by piston 40- (FIG. 2). So projected, its clamping surface 57 will be moved toward thecorresponding surface 58 of ringSl' to seal the precipitate chamber and thus isolate the solids within the same. When rotation has ceased, set screws'56may be projected to maintain clamping pressure, if the bowl assembly is to be removed from the machine for use. Sockets 55' receive the ends of a bail for carrying the container.
In FIG. 6 a container 60' has been shown to provide a bowl assembly corresponding. generally to that illustrated in FIG. 4. A ring 61 has screw-threaded connection with a bowl 62. The base of the latter is formed with openings through which rods 63 slidably project; The outer or upper ends of the rods bear against a bezel 64. The latter encircles a clamping and supporting member 65. If desired, a common ring 66 may support all the posts or rods 63, so that they will be projected as a unit; Ring 61 may be formed with openings 67 corresponding to those heretofore described and within which the preferredstoppers 49 are disposed.
A device such as is illustrated in FIG. 6 is especially valuable when used in laboratory work. It will usually have a capacity of from 4 to 10 cc. The volume of the main chamber of the container 60'may, for example, be 6 cc., and the precipitate-chamber, as defined by the annulus, in expanded condition conveniently involves cc. When the bezel- 64 has beenprojected, thecapacity of this solid-receiving chamber is conveniently reduced to 2 cc. or less.
As in FIG. 7, an apparatus is furnished with which the receptacle may be associated after its removal from the centrifuge. This apparatus will assure a ready collapse or expansion of the container 68. While no clamp has been shown in association with the latter for the purpose of sealing the precipitate-receiving chamber of the container, it will be understood that such a seal may be associated therewith or incorporated therein by any suitable expedient, as a-fore suggested. Of course, in many instances, and under conditions of filling, no seal will be necessary. In any event, a ring 69 may bear against the extended portion 70 of the receptacle. This extended portion may incorporate the heat-sealed zone of the container which maintains the sections against displacement with respect to each other. Preferably having screw-threaded attachment to ring 69 is a bowl 71. From the latter, a conduit 72 extends through to a suitable apparatus (not shown) whereby fluids under pressure may be introduced, or a condition of vacuum may be created within the space between bowl 71 and the container. With pressure introduced into that space, the base of the container will flex upwardly, as shown in full lines. With a condition of less than atmospheric pressure, the base will flex outwardly, as indicated in dot-and-dash lines. Thus, the container may have its contents expelled, or else an aspiraring action may be resorted to which will fill the container with a desired liquid.
To briefly recapitulate the sequence of operations as heretofore traversed in connection with FIGS. 1, 2 and 3, it will be understood that after the receptacle is associated with the head of the machine as defined by the ring 10 and bowl 11 thereof, the auxiliary or annular portion of the receptacle has its interior in communication with the main part of the cavity defined by the receptacle. Through the limited-area passage connecting these cavities, the solids will pass as the head rotates at high speed, to be retained within the annulus 28. If a structure such as bezel 35 is employed, then it is feasible to displace any excess serum or plasma deposited within the annulus 28 back into the main cavity of the receptacle. In any event, when the separation of the components has been effected within the limits of the machine design involved, the cooperating clamping portions 32 and 33 will be caused to seal off one cavity from the other, to thus 6 entrap the different material in those cavities. The difference between the molecular weights of the substances will assure that all the solids are within the annulus, while the serum or plasma is contained within the main body of the receptacle.
While the receptacle in its upper, intermediate and lower sections should include a certain amount of flex ibility, it is preferred that at least certain of these parts be sufiiciently firm to embody stability in the receptacle as a whole. Also, it is preferred that the receptacle be provided with a seal of the type shown at 26, 27, 28' and 29, rather than have a permanently open upper face. The attachment of ring 10 to bowl 11 may be effected in-many different manners to expose the cavities within the head. It is apparent that while the present invention is of great value when applied to the separation of blood components, itmay be advantageously employed for the separation of materials having different specific gravitiesi In many instances it will be desirable to employ liquid; as indicated in FIGS. 2 and 3, within bowl 11 for the purpose of assuring :a proper balance or equilibrium in the assembly. However, the apparatus will function entirely satisfactorily in the absence of that liquid, if the speed of operation isheld within design limitations.
Thus, among others, the several objects of theihvention :as specifically aforenoted are achieved. It is apparent that the construction of the machine and receptacle may be revised in numerous respects without departing from the spirit of the invention as defined by the claims.
1. Apparatus for use in separating materials into components of relatively higher and relatively lower specific gravity which comprises: a drive shaft and IE1 motor for causing the rotation thereof, a rotor head operatively connected to the drive shaft to be rotated thereby and formed With a circular recess concentric with the center of rotation, a separate removable liner in the recess in the rotor head in the form of a hollow circular body having upper and lower walls of sheet material spaced from each other to provide a central chamber and connected together around their peripheries to provide an annular chamber closed around the outer edge and communicating with the central chamber at its inner edge, said walls having annular confronting sealing zones between the two chambers, at least one of the walls being flexible so that sealing zones can be brought into sealing engagement with each other, said rotor having clamping means spaced inwardly from the periphery of the recess in registry with the sealing zones of the liner to clamp said sealing zones into sealing engagement with each other, means for compressing the flexible wall of said liner around the annular chamber while the rotor head is rotating to express some or" the materials from the annular chamber into the central chamber and means for thereafter operating said clamping means while said rotor head is rotating to cause said sealing zones to have sealing engagement and thereby seal off the materials of relatively higher specific gravity in the annular chamber from the materials of relatively lower specific gravity in the central chamber.
' 2. Apparatus for use in separating materials into com ponents of relatively higher and relatively lower specific gravity comprising a drive shaft and a motor for causing the rotation thereof, a rotor head operatively connected to the drive shaft to be rotated thereby and formed with a circular recess concentric with the center of rotation, a separate removable liner in the recess in the rotor head in the form of a hollow circular body having upper and lower walls of sheet material spaced from each other to provide a central chamber and connected together around their peripheries to define an annular chamber closed around the outer edge and communicating with the central chamber at its inner edge, said walls having .annular confronting sealing zones between the two chambers,
at least one of the walls being flexible so that sealing zones can be brought into sealing engagement with each other, said head having clamping means space-d inwardly from the periphery of the recess in registry with the sealing zones of the liner to clamp said sealing zones into sealing engagement with each other, and means for operating said clamping means While said rotor head is rotating to cause said sealing zones to have sealing en gagernent and thereby sealing off materials of relatively higher specific gravity in the annular chamber from materials of relatively lower specific gravity in the central chamber.
3. A receptacle for use as a disposable liner in a centrifuge rotor of the type which during rotation seals off one annular area thereof to segregate material of relatively higher specific gravity from material of relatively lower specific gravity, said receptacle comprising: a hollow circular body having an upper wall and a lower wall made of sheet material spaced from each other to provide a central chamber and being connected together continuously around the periphery of the body to provide an annular chamber extending completely around the body, said annular chamber being enclosed around its outer edge and communicating with the central chamber along its inner edge, said walls having annular sealing zones extending around said body between said chambers in confronting relationship with each other, with at least one of said Walls being flexible so that the sealing zones can 8, be brought into sealing engagement with each other to seal off the annular chamber from the central chamber and said upper wall being provided with a central portion through which materials separated in the receptacle can be withdrawn.
4. A receptacle for use as a disposable liner in a centrifuge rotor of the type which during rotation seals ofi one annular area thereof to segregate material of relatively higher specific gravity from material of lower specific gravity as set forth in claim 3 in which the lower wall of the receptacle body is flexible and the upper wall is relatively more rigid.
5. The invention in accordance with claim 3 wherein access means are on said receptacle at said annular chamber for providing access to the annular chamber.
References Cited in the file of this patent UNITED STATES PATENTS 2,023,762 Fawcett Dec. 10, 1935 2,272,675 Knudsen Feb. 10, 1942 2,656,263 Larsen Oct. 20, 1953 2,883,103 Whitehead et al Apr. 21, 1959 2,898,037 Dega Aug. 4, 1959 FOREIGN PATENTS 66,298 Sweden Oct. 30, 1928 252,907 Germany Oct. 28, 1912 971,978 France Aug. 23, 1950