US3352486A - Apparatus for washing particle matter - Google Patents

Description

NOV. 14, 1967 51555 3,352,486

APPARATUS FOR WASHING PARTICLE MATTER Filed A ril 30, 1965 2 etsh 1 VACUUM SOURCE .9

mygmoa Mary B. Gibbs ATTORNEY Nov. 14, 1967 M. B. GIBBS 3,352,436

APPARATUS FOR WASHING PARTICLE MATTER.

Filed April 30, 1965 -2 Sheets-Sheet 2 5 FIG.3

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ATTORNEY United States Patent 3,352,486 APPARATUS FOR WASHING PARTICLE MATTER Mary B. Gibbs, Hyattsville, Md., assignor to Clay-Adams, Inc., New York, N.Y., a corporation of New York Filed Apr. 30, 1965, Ser. No. 452,320 6 Claims. (Cl. 233-26) This invention relates to methods and apparatus for washing particle matter; and more particularly concerns procedures and means for washing erythrocytes and other formed elements of the blood free of contaminating serum proteins or other material.

conventionally, erythrocytes are washed by first centn'fuging the blood sample in a test tube to pack the red cells, decanting the supernatant plasma or serum, adding washing solution, inverting and shaking the test tube manually, centrifuging and decanting. This may be repeated, with subsequent additions of wash solution, several times, to achieve the desired degree of removal of contaminating agents. However, contamination may occur due to the tendency of droplets to cling to the test tube wall, leaving residual matter on the test tube wall. Also, decantation may deliberately be incompletely carried out in order to avoid loss of cells.

An object of this invention is to provide improved methods and apparatus for washing centrifuged blood samples and eliminating the temptation to seal test tubes with finger tips while resuspending the sediment. The latter practice is known to cause cross contamination from specimen to specimen.

Another object of this invention is to reduce the danger of loss of sediment when supernatant material is hastily decanted.

A further object of this invention is to provide improved apparatus for simultaneously washing a large number of blood samples in individual test tubes while such tubes are still mounted in a centrifuge, after the centrifuging operation, thereby avoiding inadvertent mistakes in identification of the individual specimens, not uncommon in known manual washing procedures.

Another object of this invention is to provide improved transfer means for supplying washing solutions to blood samples carried in test tubes before centrifugation, the transfer means also serving to remove supernatant material after centrifugation of the samples; the transfer means further serving to supply Washing solution in a manner to dislodge the cells from the tube walls and to carry the formed elements into the upper portions of the tubes.

A further object of this invention is to provide manifold type, improved means for injecting washing solution into test tubes carrying blood samples and for removing super natant material without decantation so as to materially reduce danger of cellular loss when the washing operation is repeated; I

Still another object of this invention is to provide improved washing apparatus in manifold form for ejecting washing solution into individual test tubes carrying centrifuged blood specimens, the ejected stream of washing solution being located and directed in a manner to dislodge the washed, centrifuged pack of red cells from the tube wall in an efiicient manner.

Known, manual procedures for washing cells does not permit standardization of the cell washing operation. Accordingly, yet another object of this invention is to provide a cell washing procedure and apparatus which permits standardization of the same, thereby producing a washed cell residue of a given dryness.

Other objects of this invention will in part be obvious and in part hereinafter pointed out.

In the drawings:

FIG. 1 is a side elevational view of apparatus for washing blood samples and embodying the invention herein;

FIG. 2 is a perspective view of the sample carrying tubes disposed in the centrifuge basket;

FIG. 3 is a perspective view showing the manifold Vdevice for ejecting and aspirating washing solution, embodying the invention;

" stant invention comprises a manifold head arranged for connection to a source of washing solution which may be delivered to the head under a given'pressure for distribution to individual dispensing tubes conveniently arranged in a manner, whereby the dispensing tubes may be inserted into the test tubes carrying the blood specimens to be washed as said test tubes are disposed in a suitable arrangement on the head of a conventional centrifuge. The manifold head may also be subjected to negative pressures whereby supernatant material from the test tubes may be aspirated without unduly disturbing the centrifuged deposit of red cells. Also, the dispensing tubes extending from the manifold head are so located in respect to the test tube walls as to effectively dislodge the centrifuged deposit of cells, when further washing solution is passed to the dispensing tubes under pressure.

Thus, as shown in FIG. 1, 10 designates a conventional centrifuge of the type used in serology and which carries within the upper portion thereof the usual test tube basket 11, see FIG. 2, which includes slotted discs 12, 13 for locating the individual test tubes T, which carry the blood specimens to be washed, in circular arrangement. The discs 12, 13 are formed with radial slots 14, 15 respectively to allow the tubes T to assume inclined positions during the centrifuging operation and to be disposed vertically before and after such centrifuging operation.

Means is provided for delivering washing solution, such asv the usual saline solution, simultaneously to each test tube T while such tubes T are still carried in basket 11 in centrifuge 10. To this end, there is provided a manifold head 16 in the form of a flat circular hollow member having a top wall 17 and having a centrally located opening 18 therein from which extends an upstanding connector, tubular member 19, and a bottom wall 20, thus forming a circular chamber 21 which is in communication with tube 19. The bottom wall 20 of chamber 21 is formed with a circular arrangement of openings 22 disposed adjacent the periphery thereof. Small bore tubings 23 are fitted at their upper ends into openings 22 and are thus in communicating relation with chamber 21.

The head 16 is adapted to be disposed in aligned relation to test tubes T so as to locate each tubing 23 over a test tube T, for insertion and withdrawal relative thereto. When tubings 23 are fully inserted into test tubes T, the tips thereof are located adjacent the lower ends of the test tubes.

A collar 24 formed with through openings 24A arranged in a circle is adapted to be slidably mounted on tubes 23, chain means 25 interconnecting head 16 with collar 24 to limit the lowered movement of said collar on tubings 23.

As shown in FIG. 1, upstanding tube 19 on head 16 is adapted to be connected to a supply of washing solution as may be carried in a container 26, by way of a flexible tubing 27, means for delivering measured quantities of washing solution, such as saline, as by a hypodermic syringe 28 and vflexible tubing 29 connected to connector member 19.

In use, the manifold head 16 is located over the test tubes T while they are in their vertical positions in centrifuge the movable collar 24 facilitating the proper vertical positioning of tubings 23 for entry into the respective test tubes T.

Syringe 28 is operated to withdraw a selected amount of washing solution from container 26 to chamber 21 of the head 16 and thence to tubings 23.

The manifold 16 is then removed from its dispensing position to allow the cover of centrifuge 10 to be closed and the centrifuge to be operated in the usual manner. After centrifugation has been completed, supernatants in each of test tubes T may be removed without decantation, by means of a second manifold head 16A entirely similar to that previously described, except that its flexible tubing 29A is connected to a source of reduced pressure as a vacuum pump 30 and a receiver 31.

The blood samples in test tubes T may be repeatedly washed with saline washing solution from container 26 by repeating the operations described above; the syringe 28 being retracted to fill the same with new charges of washing solution in measured amount which is ejected into dispensing tubes 23 when the plunger of syringe 28 is pressed forwardly.

When the supernatants are aspirated out of the test tubes T after the previous washing operation, fresh washing solution is ejected from tubings 23 by appropriate movement of the syringe plunger and the deposited red cells C or other formed elements collected on the outer wall portion 32 or bottom wall portion 33 of each of the test tubes T due to successive centrifugation, will be dislodged and carried to the upper portions of tubes T in the wash solution. The washing cycle may be repeated two or more times with intermediate short period centrifugation. The washed cells may then be used in conventional test procedures.

The forced concentration of formed elements in the upper portion of test tubes T is an extremely effective washing operation inasmuch as the greater distance be tween the centroid of the formed elements and the bottom of said test tubes allows for more contact between the washing solution and each particle during centrifugation.

It is understood that the Washing operation may be carried out with a single manifold head 16, using the same for dispensing washing solution and for aspirating supernatants in alternation. However, in such case, the head 16, its dispensing tubes 23 and connecting flexible tubing 29 are well rinsed between such operations to avoid any possibility of contamination.

While the manifold head 16 and its tubes 23 are shown as particularly adapted for use with specimen tubes normally in a vertical position, it is understood that such head may be modified to dispose its dispensing-aspirating tubes at a selected angle to correspond to the disposition of test tubes arranged at a corresponding angle.

While the apparatus and process herein disclosed is shown in connection with the washing of erythrocytes, it is understood that the same may be used in washing other particle matter such as latex, bentonite, bacteria, spores, tissue cells, chemical and bacteriological precipitates.

What is claimed is:

1. Cell washing means for use with a plurality of test tubes, each test tube carrying a centrifuged blood specimen portion deposited at the lower end thereof, said 4 washing means comprising means for supporting said test tubes in an upstanding arrangement, manifold means movable towards and away from said supporting means, said manifold means comprising a plurality of transfer tubings depending therefrom and in communication therewith, said transfer tubings being in a vertical arrangement, said manifold means being movable toward said supporting means with said transfer tubings in respective alignment with said supported test tubes for introducing said transfer tubings into the respective test tubes, said tubings having a length to locate the lower ends thereof at a point within said test tubes adjacent the deposit therein, said manifold means including inlet means for admitting washing solution therein and the distribution thereof via said transfer tubings to the lower end portion of each test tube for washing the deposit therein.

2. Cell washing means as in claim 1 wherein said inlet means on the manifold means is adapted to be con.- nected to a source of reduced pressure whereby to aspirate supernatants from each of said test tubes by way of said transfer tubings and passages in said manifold means.

3. Cell washing means as in claim 1 wherein each of said transfer tubings comprises small bore tubing, and collar means having a plurality of through openings arranged for passing the respective tubings therethrough, said collar means being slidably mounted on said tubings for movement to positions to properly locate the lower portions of said tubings for entry into the respective test tubes.

4. For use with test tubes carrying blood specimens to be subjected to washing solutions and centrifugation to effect the separation of liquid from the formed elements thereof, said test tubes being in a centrifuge head, improved cell washing means comprising a pair of similar manifold heads, movable towards and away from said centrifuge heads, each manifold head comprising a chamber and inlet means communicating with said chamber, and a plurality of transfer tubings mounted on said manifold head, said tubings communicating at one end thereof with said chamber, the arrangement of tubings on said manifold heads being correlated to the arrangement of the test tubes on said centrifuge to allow the tubings to be respectively inserted into the test tubes with each tubing lying within the test tube associated therewith, said tubings having a length to locate the outer ends thereof adjacent the lower ends of said test tubes when said tubings are fully inserted into said test tubes, one of said heads having the inlet means thereof arranged for connection to a source of washing solution, the other of said heads having the inlet means thereof arranged for connection to a source of reduced pressure whereby said heads may be used in succession to (1) inject washing solution into each of said test tubes; and (2) to aspirate supernatants from each of said test tubes.

'5. Cell washing means as in claim 4 wherein the tubings on said manifold head are in a circular arrangement.

6. In combination with a manifold comprising a chamber portion inlet means on said manifold in communication with said chamber portion and a plurality of flexible transfer tubings mounted on said manifold and in communication with the chamber portion thereof, said tubings being arranged in a predetermined pattern and in parallel relation to each other, means for maintaining the outer ends of said tubings in parallel relation to each other comprising collar means having a plurality of through openings of a diameter for slidably passing the respective tubings therethrough, whereby said collar means may be slidably moved relative to said tubings toward the outer ends thereof to bring the outer ends of said tubings into parallel relation to each other.

(Other references on following page) 6 References Cited 3,202,348 8/1065 Strohmaier 23326 UNITED STATES PATENTS 3,208,468 9/1965 Hansen 137-266 1/1937 Lux 23272.7 1 014 348 8 igfi Gq PATENTS 8/1956 Hatcher 239 229 5 ermany' 10/ 1958 Snyder 23-272] HENRY T. KLINKSIEK, Primary Examiner.

2/1965 Berg 239--229 X M. CARY NELSON, Examiner.

Claims (1)

1. CELL WASHING MEANS FOR USE WITH A PLURALITY OF TEST TUBES, EACH TEST TUBE CARRYING A CENTRIFUGED BLOOD SPECIMEN PORTION DEPOSITED AT THE LOWER END THEREOF, SAID WASHING MEANS COMPRISING MEANS FOR SUPPORTING SAID TEST TUBES IN AN UPSTANDING ARRANGEMENT, MANIFOLD MEANS MOVABLE TOWARDS AND AWAY FROM SAID SUPPORTING MEANS, SAID MANIFOLD MEANS COMPRISING A PLURALITY OF TRANSFER TUBINGS DEPENDING THEREFROM AND IN COMMUNICATION THEREWITH SAID TRANSFER TUBINGS BEING IN A VERTICAL ARRANGEMENT, SAID MANIFORLD MEANS BEING MOVABLE TOWARD SAID SUPPORTING MEANS WITH SAID TRANSFER TUBINGS IN RESPECTIVE ALIGNMENT WITH SAID SUPPORTED TEST TUBES FOR INTRODUCING SAID TRANSFER TUBINGS INTO THE RESPECTIVE TEST TUBES, SAID TUBINGS HAVING A LENGTH TO LOCATE THE LOWER ENDS THEREOF AT A POINT WITHIN SAID TEST TUBES ADJACENT THE DEPOSIT

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