U.S. patent number RE38,757 [Application Number 09/482,653] was granted by the patent office on 2005-07-12 for automatic multiple-decanting centrifuge and container therefor.
This patent grant is currently assigned to Harvest Technologies Corporation. Invention is credited to Steven M. Gann, Lin A. Jakary, John R. Wells.
United States Patent |
RE38,757 |
Wells , et al. |
July 12, 2005 |
Automatic multiple-decanting centrifuge and container therefor
Abstract
A centrifuge is capable of holding a sample container in
selected orientations, either during or after centrifugation, to
drain supernatants between two or more chambers of the container.
The draining may be gravity or centrifugal draining. This allows an
automated process to subject a sample to a first physical or
chemical treatment to produce a first supernatant, the first
supernatant to be subjected to a second physical or chemical
treatment, and a second supernatant to be separated from a desired
component.
Inventors: |
Wells; John R. (late of Culver
City, CA), Jakary; Lin A. (La Jolla, CA), Gann; Steven
M. (Huntington Beach, CA) |
Assignee: |
Harvest Technologies
Corporation (Plymouth, MA)
|
Family
ID: |
23729282 |
Appl.
No.: |
09/482,653 |
Filed: |
January 13, 2000 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
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Reissue of: |
435662 |
May 5, 1995 |
05707331 |
Jan 13, 1998 |
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Current U.S.
Class: |
494/20; 220/501;
494/32 |
Current CPC
Class: |
B04B
5/0421 (20130101); B04B 9/14 (20130101) |
Current International
Class: |
B04B
5/00 (20060101); B04B 5/02 (20060101); B65D
41/50 (20060101); B65D 41/32 (20060101); B04B
005/02 (); B65D 041/50 () |
Field of
Search: |
;494/13,14,16,17,20,32,33,35,85 ;210/781,782 ;215/6 ;422/102
;220/501,507,523,554 |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
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461698 |
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Dec 1949 |
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CA |
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4323844 |
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Dec 1949 |
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DE |
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43 23 844 |
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Jul 1993 |
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DE |
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936560 |
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Jul 1948 |
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FR |
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Primary Examiner: Cooley; Charles E.
Attorney, Agent or Firm: Clark & Brody
Claims
We claim:
1. A centrifuge comprising means for removably receiving a unitary
container having a plurality of chambers for receiving substances
to be centrifuged, means for rotating said container to subject
said substances to centrifugation, and means for locking said
container in a first predetermined position to allow a supernatant
in a first of said chambers to transfer into a second of said
chambers and for locking said container in a second position to
transfer a supernatant in said second chamber to another of said
chambers.
2. Apparatus according to claim 1 wherein said means for locking,
when activated, locks said container such that a supernatant in one
of said chambers transfers into another of said chambers by gravity
draining.
3. Apparatus according to claim 1 wherein said means for locking,
when activated, locks said container such that a supernatant in one
of said chambers transfers into another of said chambers by
centrifugal transferring.
4. Apparatus according to claim 1 wherein said means for locking,
when activated to a first position, locks said container such that
a supernatant in said first chamber drains into said second chamber
by gravity draining and, when activated to a second position, locks
said container such that a supernatant in said second chamber
transfers into said first chamber by centrifugal transferring.
5. Apparatus according to claim 1 wherein said locking means
comprises a movable plate and means for controlling the position of
said plate.
6. Apparatus according to claim 5 wherein means for controlling is
electrical.
7. Apparatus according to claim 6 wherein said means for
controlling is magnetic.
8. Apparatus according to claim 1 further comprising means for
controlling said means for locking and said means for rotating to
provide automatic multiple decanting by activating said means for
rotating for a predetermined period of time, activating said means
for locking to allow a supernatant in said first chamber to
transfer into said second chamber, activating said means for
rotating a second time, and activating said means for locking a
second time to allow a supernatant in said second chamber to
transfer into said first chamber.
9. Apparatus according to claim 8 wherein said means for locking
locks said container such that a supernatant in said first chamber
transfers into said second chamber by gravity draining and locks
said container such that a supernatant in said second chamber
transfers into said first chamber by centrifugal transferring.
10. Apparatus according to claim 1 further comprising means for
controlling the temperature of the contents of said second
chamber.
11. Apparatus according to claim 10 wherein said means for
controlling the temperature is capable of freezing said contents
for cryoprecipitation.
12. Apparatus for separation of a precipitate from a liquid
comprising a unitary container having first and second adjacent
chambers, wherein said first chamber is located with respect to
said second chamber such that a first supernatant in said first
chamber drains by gravity into said second chamber when said first
and second chambers are held in a first orientation and a second
supernatant in said second chamber transfers from said second
chamber into said first chamber by centrifugal transferring when
said first and second chambers are held in a second orientation and
subjected to centrifugation.
13. Apparatus according to claim 12 wherein said first and second
chambers are joined by a wall that forms a fluid flow path between
said first and second chambers.
14. Apparatus according to claim 13 further comprising divider
means for dividing said first chamber into two .[.pads.].
.Iadd.parts.Iaddend., said divider means being located near the
expected location of the interface between said precipitate and
said liquid.
15. Apparatus according to claim 14 wherein said divider means
includes a periphery having at least one groove therein for
allowing fluid communication between said two parts.
16. Apparatus according to claim 12 further comprising a covering
on said first and second chambers for preventing spillage of the
contents of said chambers while allowing a syringe to inject fluids
into or remove fluids from said chambers.
17. Apparatus according to claim 16 wherein said covering includes
access port means for each of said chambers for allowing a fluid to
be introduced into a chamber and means for sealing said access port
means until opened to allow said fluid to pass.
18. Apparatus according to claim 17 wherein at least one of said
chambers includes a hollow tube aligned with a said access port for
conducting said fluid into said at least one of said chambers.
19. Apparatus according to claim 18 further comprising air vent
means for allowing air in said container to exit from said
container.
20. Apparatus according to claim 12 in combination with a
centrifuge for subjecting said liquid to centrifugation, locking
said chambers in said first orientation to allow said first
supernatant to drain into said second chamber, and locking said
chambers in said second orientation while rotating said chambers to
provide said centrifugal transferring.
21. A centrifuge comprising a first chamber for receiving a fluid
substance and a second chamber for receiving a fluid substance,
means for rotating said first and second chambers to subject said
substances to centrifugation, and means for locking said chambers
in first predetermined positions and for locking said chambers in
second predetermined positions, means for transferring a
supernatant in said first chamber into said second chamber by
gravity when said chambers are in said first predetermined
positions and for transferring a supernatant in said second chamber
to said first chamber by centrifugal transfer when said chambers
are in said second predetermined positions..Iadd.
22. A system for treating physiological products, comprising: a
centrifuge; a container having at least a first chamber and an
adjacent second chamber, wherein each of the first and second
chambers has a top portion, a bottom portion and a set of walls,
wherein the top portions of the first chamber and second chamber
are adjacent each other and connected by a bridge that transfers
fluid therebetween when said container is in a predetermined
orientation; and a holder assembly attached to the centrifuge and
effective to removably receive the container and orient the
container in said predetermined orientation..Iaddend..Iadd.
23. The system of claim 22, wherein the chambers include removable
lid portions, thereby forming a closed
container..Iaddend..Iadd.
24. The system of claim 23 wherein at least one of the chambers
includes an access port for transference of a
liquid..Iaddend..Iadd.
25. A container comprising: a first sterile chamber having a first
top portion, a first bottom portion and a first set of walls; a
second sterile chamber adjacent said first sterile chamber and
having a second top portion adjacent said first top portion, a
second bottom portion and a second set of walls; a bridge
connecting said first top portion of the first chamber and said
second top portion of the second chamber, such that a liquid can be
transferred from the first chamber to the second chamber while the
container is positioned at a predetermined angle, and means for
sterile transfer of a liquid to or from at least one of said
chambers independently of the other of said chambers and located
near the top of at least one of said chambers..Iaddend..Iadd.
26. The container of claim 25, wherein the chambers include a
removable lid portion..Iaddend..Iadd.
27. A system for treating physiological products and maintaining
sterility of said products during said treating comprising: a
container having a plurality of closed, sterile fluid-receiving
chambers, a bridge forming a fluid path allowing fluid
communication between a first of said chambers and a second of said
chambers when said container is in a predetermined orientation, and
at least one access port allowing sterile access to at least one of
said chambers, and a centrifuge having a holder removably receiving
said container and allowing said container to assume a first
orientation wherein a physiological product in one of said chambers
is subjected to centrifugation and said predetermined orientation
wherein fluid in said first of said chambers flows along said fluid
path to said second of said chambers and said centrifuge comprises
a locking element that selectively holds said container in said
predetermined orientation..Iaddend..Iadd.
28. A system according to claim 27 wherein said holder comprises a
frame pivotally mounted to a rotor of said
centrifuge..Iaddend..Iadd.
29. A system according to claim 27 wherein said locking element
comprises a movable locking plate that is movable between free and
locking positions, wherein said plate allows said container to
assume said first orientation when in said free position and holds
said container in said predetermined position when in said locking
position..Iaddend..Iadd.
30. A system according to claim 29 further comprising an
electromagnet for moving said locking plate to one of said locking
and free positions..Iaddend..Iadd.
31. A system according to claim 27 wherein said holder comprises a
frame pivotally mounted to a rotor of said centrifuge, and said
locking element comprises a movable locking plate that is movable
between free and locking positions, wherein said movable locking
plate engages said frame to allow said container to assume said
first orientation when in said free position and to hold said
container in said predetermined position when in said locking
position..Iaddend..Iadd.
32. A container comprising a base forming a plurality of sterile
chambers, each of said chambers having a bottom and a top, a bridge
connecting top portions of at least two of said chambers and
arranged to provide a sterile fluid channel from a first of said at
least two sterile chambers to a second of said at least two sterile
chambers when said container is in a predetermined orientation, a
lid closing said top of each of said plurality of chambers, and an
access port near the top of at least one of said chambers forming
an opening covered by an element that allows sterile transfer of a
liquid through said opening to or from said at least one of said
chambers independently of the other of said
chambers..Iaddend..Iadd.
33. A container according to claim 32 wherein said plurality of
sterile chambers and said bridge comprise a molded base
part..Iaddend..Iadd.
34. A container according to claim 33 wherein said container is
substantially rigid..Iaddend..Iadd.
35. A container according to claim 32 further comprising a
separation disk in one of said chambers..Iaddend..Iadd.
36. A container according to claim 32 wherein said plurality of
chambers comprise first and second adjacent chambers having
adjacent sidewalls and said bridge is formed at the tops of said
adjacent sidewalls..Iaddend.
Description
TECHNICAL FIELD
This invention relates to the art of automatic centrifugation. In
particular, the invention relates to apparatus and procedures using
automatic, multiple decanting with centrifugation. In a preferred
embodiment, an automated procedure separates blood components and
proteins including the separation of fibrinogen from blood.
BACKGROUND
The separation of components through centrifugation is well known.
For example, in the medical field it is common to subject a sample
of blood to centrifugation to produce a precipitate of cellular
material and a supernatant of plasma. The plasma is then decanted
to complete the separation of these components.
U.S. Pat. No. 5,178,602 (Wells) and U.S. Pat. No. 5,047,004 (Wells)
show an automated centrifuge, which includes structure for holding
a centrifuge tube, after centrifugation, in a position that allows
the supernatant to drain from the tube and into another container
by gravity. The holding structure shown in these patents comprises
a locking mechanism mounted for axial movement with respect to the
axis of rotation of the centrifuge. An electromagnet that is easily
controlled causes the axial movement.
It is also known to decant a supernatant by the process of
centrifugal draining. According to that process, a centrifuge
rotates a centrifuge tube while the tube is held in a position such
that the supernatant is drained from the tube by centrifugal
forces.
Fibrin sealants for treating wounds are known and are typically
produced by combining a fibrinogen/Factor XIII component with
bovine thrombin. When these are mixed, a fibrin tissue adhesive
results, which is applied to the wound. Descriptions of
compositions for use as tissue sealants are given in U.S. Pat. No.
5,292,362 and U.S. Pat. No. 5,209,776 (Bass et al.). The fibrinogen
is obtained from plasma, either pooled or autologous, and
cryoprecipitation is one known technique for separating fibrinogen
from plasma. One cryoprecipitation technique is described in U.S.
Pat. No. 5,318,524 and includes the centrifugation of thawing
plasma to produce a precipitate containing fibrinogen/Factor XIII.
Other techniques for producing fibrinogen/Factor XIII include
inducing precipitation of the component by addition of such agents
as Ammonium Sulfate or polyethylene glycol (PEG) to blood
plasma.
SUMMARY OF THE INVENTION
Several known chemical procedures include repeated steps of
physical separation between two or more components. Separation
based on density differences between the components is often by
centrifugation, and the resulting supernatant is decanted to
complete the separation. Each step provides an opportunity for
error, which would be reduced by automation of the process.
In accordance with the invention, chemical procedures requiring
several centrifugation steps are automated, to reduce the time
required by a clinician and eliminate the potential for errors.
Apparatus in accordance with the invention includes a
multiple-chamber container and a centrifuge designed to receive the
container and subject its contents to predetermined centrifugation
steps as well as gravity and centrifugal decanting of the
supernatant.
A preferred container in accordance with the invention includes
first and second chambers separated by an intermediate wall. The
first chamber is designed to receive a first liquid, such as human
blood. The second chamber is located adjacent the first chamber,
and the wall between the chambers is such that a supernatant in the
first chamber will flow over the top of the wall and be drained
into the second chamber by gravity when the container is held in
the proper orientation. The supernatant in the second chamber may
then be subjected to a mixing action and then may be subjected to a
second centrifugation. The container can also be held in a second
position whereby a second supernatant is caused to flow back over
the wall into the first chamber by centrifugal forces resulting
from a second centrifugation.
A centrifuge in accordance with the invention includes a rotatable
support with a swinging frame for receiving the multiple-chamber
container and means for locking the container in either of at least
two positions for draining supernatant fluids from the chambers.
Preferably, the locking means is an electro-magnetically operated
disk mounted for movement axially with respect to the axis of
rotation of the rotatable support. The centrifuge is preferably
operated under the control of an electronic circuit, which may
include a programmed array logic (PAL) or other circuitry, that
causes the rotor to operate in accordance with a predetermined
program and controls the locking means such that it locks the
container in predetermined orientations in conjunction with
operation of the rotor.
While many different programs for operation of the centrifuge can
be developed, depending on the desired results, a preferred
operation is for the production of autologous fibrinogen. Prior
techniques for production of fibrinogen require several distinct
steps, each of which requires a skilled technician but does not
eliminate an opportunity for error. These steps include separation
of plasma from cellular components, treatment of the plasma with a
precipitating agent, and separation of a fibrinogen precipitate
"pellet" from the plasma. The separation of plasma from blood and
the separation of the fibrinogen pellet from plasma typically
require centrifugation first of the blood and then of the plasma,
with addition of at least one precipitating agent between the
steps. Thus, the production of fibrinogen in the prior art has been
complex and error-prone.
In accordance with this embodiment of the invention, a patient's
anticoagulated blood is placed in the first chamber of the
disposable container, and a precipitation agent is placed in the
second of the chambers. The container is then placed in the
swinging frame of the centrifuge, and the control circuit is
activated to initiate the operation of the centrifuge. The
centrifuge first rotates the container for a time period that has
been determined to be adequate for separating the cellular
components from the supernatant plasma. During this time, the
swinging frame will have rotated outwardly substantially due to
centrifugal forces on the container. While the frame is in the
outwardly rotated position, the locking means is activated to lock
it there. The rotation of the support is then terminated. As the
rotational velocity of the support decreases, the supernatant
fluid, being no longer subject to the centrifugal forces, flows out
of the first chamber and into the second chamber by gravity. The
cellular component is more viscous and, thus, flows toward the
second chamber at a rate less than that of the plasma. Preferably,
however, a divider in the form of a disk is placed in the first
chamber to restrict the flow of the cellular components and plasma
below the disk. The disk is at a depth that provides a
predetermined volume of plasma, which is normally near the expected
boundary between the supernatant and cellular components. After a
period of time that has been determined to allow an adequate amount
of the plasma to flow into the second chamber, the locking means is
deactivated to release the container, whereby it assumes an upright
position with the cellular component remaining in the first chamber
and the plasma now in the second chamber. The rotatable support is
then alternately activated and deactivated for short intervals to
mix the plasma with the precipitating agent in the second chamber.
Interaction between the precipitating agent and the plasma
initiates precipitation of fibrinogen and Factor XIII from the
plasma. The support is then again rotated to accelerate the
precipitation of the fibrinogen/Factor XIII and to create a pellet
in the bottom of the second chamber. As a final step, the locking
means is again activated to lock the container in a position such
that the supernatant resulting from precipitation of the fibrinogen
is decanted by centrifugal draining into the first chamber. In this
step, the container is held substantially upright, and the support
is rotated to apply centrifugal forces to the supernatant, whereby
it flows over the wall between the chambers and into the first
chamber. The locking means is then inactivated, the container
removed from the centrifuge, and the fibrinogen/Factor XIII removed
from the second chamber for further processing. In a preferred
embodiment, the fibrinogen/Factor XIII is reconstituted and then,
combined with thrombin, and applied to a patient to treat a
wound.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is a perspective of a container and centrifuge in accordance
with the invention.
FIG. 2 is a vertical cross section of a preferred embodiment of a
container.
FIGS. 3a and 3b are partial vertical cross sections of the
centrifuge of FIG. 1.
FIGS. 4a through 4f are schematic diagrams illustrating a preferred
method of operation of the centrifuge of the invention.
DETAILED DESCRIPTION OF THE INVENTION
With reference to FIGS. 1 and 2 of the drawings, a centrifuge 2 is
designed to receive a container 4 in accordance with the invention.
The centrifuge is capable of subjecting the container to a series
of steps that will be described in detail below. The container
includes at least two chambers, 6 and 8. Chamber 6 is designed to
receive a first fluid to be treated, such as blood. Chamber 8 is
designed to receive fluids that have been decanted from chamber 6,
such as a supernatant plasma resulting from centrifugation of blood
in chamber 6.
A preferred form of the container is shown in detail in FIG. 2. As
shown, the container comprises three primary parts. A base part is
preferably molded and includes the chambers 6 and 8 and a bridge 7,
which connects the two chambers. A lid 11, also preferably molded,
fits over the tops of the chambers to close them. The lid includes
cup shaped extensions 12 and 14, each of which is centrally aligned
with a respective one of the chambers 6 and 8. Extension 12 has a
access port in the form of centrally located opening 13, while
extension 14 has a centrally located opening 15. The openings
receive syringe needles to permit fluids to be injected into the
chambers or withdrawn therefrom. Membranes 16 and 17 cover the
openings 13 and 15 to maintain sterility. The membranes are
preferably heat sealed into the extensions 12 and 14 during
construction by providing a cavity for receiving the membranes.
After a membrane is inserted, the upper edges of the cavity are
folded over and welded, e.g., ultrasonically, to retain the
membrane.
The lid also includes a bridge T that cooperates with bridge 7 in
the base to form a fluid channel 18, connecting chambers 6 and 8.
As shown, the bridge 7 extends above the tops of the chambers 6 and
8 to prevent communication between the chambers by "splashing."
Intentional fluid communication between the two chambers will be
described in detail below.
A separation disk 20 is preferably placed in chamber 6 near, but
always above, the expected vertical position of the boundary
between supernatant plasma and cellular components after a first
centrifugation of a blood sample. The hematocrit is known to vary
among individuals, and the exact amount of plasma that will result
from a blood sample cannot be accurately predicted without prior
testing of the sample. Thus, disk 20 is located such that the
plasma above the disk after centrifugation of a predetermined
volume of blood is a predetermined volume of plasma. The upper
surface of the disk 20 is tapered toward an edge, and the edge
includes at least one groove 22 that allows fluid communication
between the parts of the chamber 6 that are above and below the
disk 20.
In a preferred embodiment, a cylindrical support 24 is attached to
the lower surface of the disk to set the location of the disk
during assembly.
A hollow tube 26 is provided to facilitate introduction of the
blood sample to the portion of the chamber 6 that is below the disk
20. The tube 26 extends from just below the opening 13 through disk
20. Thus, a syringe needle inserted through opening 13 pierces
membrane 16 and communicates with tube 26 to allow injection of the
blood sample into the bottom of the chamber 6. The groove 22
permits downward movement of the plasma and cellular components
during centrifugation but retards movement of the cellular
components during decanting. Also, an air vent 27 is provided for
chamber 8 to facilitate introduction and withdrawal of fluids.
In use, a container 4 is placed in a holder on the rotor of the
centrifuge as indicated in FIG. 1. To balance the rotor, two such
containers are preferably placed in the centrifuge in diametrically
opposed positions. Of course, only one container may be used and a
weight or "dummy" container used to balance the rotor.
FIGS. 3a and 3b are partial cross sections of a preferred
embodiment of a centrifuge showing the container locked in two
different positions. A rotor shaft 28 is connected to a motor (not
shown), which rotates the shaft. A rotor 30 is mounted to the shaft
for rotation and has a frame 32 pivotally mounted to the rotor 30
at pivot connection 34. The top surface (not shown) of the frame 32
has two circular openings for receiving the chambers 6 and 8
whereby the container can be placed in the frame such that the
contents of the container will be subjected to centrifugal forces
as the rotor is rotated. A bias spring 35 ensures that the frame 32
will pivot to an upright position when centrifugation is
terminated. The frame 32 may also be shaped to reduce wind
resistance, as known in the art.
A locking plate 36 is mounted coaxially with the shaft 28 for
engaging the frame 32 to lock the container in desired
orientations. The plate and the mechanism for controlling the
positions of the plate may be the substantially the same as that
shown in my previous U.S. Pat. No. 5,178,602. For example, an
electromagnet 38 may be provided to control the position of the
locking plate by action on a permanent magnet 40, which is attached
to the locking plate.
Preferably, the electromagnet 38 and magnet 40 are positioned such
that the locking plate can be placed in either of two positions. In
a first position, shown in phantom lines, the plate does not engage
the frame 32, and the frame 32 is free to rotate about pivot 34. In
a second position, shown in solid lines at 36', the locking plate
engages one of two parts of the frame 32 to hold it in one of two
selected orientations. In the position shown in FIG. 3a, a lip of
the plate engages a protuberance 42 on the frame 32 to lock the
container in the orientation shown in FIG. 3a. In the position
shown in FIG. 3b, the plate 36 engages an upper edge of the frame
32 to lock the container in the tilted position shown in FIG. 3b.
The locking plate preferably rotates with the rotor whereby it can
be moved to engage the frame during centrifugation of the contents
of the container.
The operation of the centrifuge in a preferred embodiment of the
invention will be described with regard to FIGS. 4a through 4f. In
a first step, blood is introduced into chamber 6 of the container
through opening 13. The blood has preferably been obtained from a
patient, but it may be pooled or obtained from another. A
precipitating agent 43, e.g., PEG, is then placed in chamber 8,
preferably by injection through opening 15. The container with
blood and precipitating agent are then placed in the centrifuge for
automated operation.
In the first step of automated operation, the container is allowed
to swing freely as the blood is subjected to centrifugation. As
illustrated in FIG. 4a, the cellular component 44 of the blood will
be separated from the plasma component 46 in this step. After a
predetermined time period, e.g., five minutes, the locking plate 36
is moved to a position shown at 36' whereby the container 4 is held
in the position shown in FIGS. 3b and 4b, and rotation of the rotor
is stopped. In this position, the plasma component 46 flows through
channel 18 by the force of gravity. The chamber is held in the
position of FIG. 4b for preferably about 3 seconds, which is
adequate to allow the plasma to drain by gravity into the chamber 8
but is not so long that the more viscous cellular component 44
drains into the chamber 8. The plasma 46 and precipitating agent
43, which was previously placed in chamber 8, are now both in
chamber 8. To provide complete mixing of these fluids, the locking
plate is lowered, and the rotor is caused to accelerate and
decelerate alternately for 10-20 seconds, as illustrated in FIG.
4c. The precipitating agent causes the fibrinogen/Factor XIII to
separate from the plasma, and this separation is assisted by
centrifuging the contents of the container a second time. This
second centrifugation may be for a period of about five minutes. A
fibrinogen pellet 48 is, thus, formed in the bottom of the chamber
8, as illustrated in FIG. 4d. At this stage of the process, the
plasma supernatant 46 remains in chamber 8.
Plasma 46 is separated from the fibrinogen pellet 48 by stopping
rotation of the centrifuge rotor to allow the container to pivot to
the upright position shown in FIGS. 3a and 4e. The locking plate 36
is then activated to lock the container in that orientation by
engagement with protuberance 42, and the container is again rotated
by the rotor for a period of about three to eight seconds. This
rotation causes the supernatant plasma 46 to flow back through
channel 18 and into chamber 6 by centrifugal draining, as
illustrated in FIG. 4e. Thus, the fibrinogen pellet and plasma have
now been separated. As a final step, the container is subjected to
another centrifugation illustrated in FIG. 4f for about fifteen
seconds, whereby the fibrinogen pellet is forced into the bottom of
the chamber 8.
The automated process for production of fibrinogen is at this point
complete, and the fibrinogen pellet is preferably extracted from
the container 8 by a syringe for further processing. For example,
the fibrinogen may be reconstituted and combined with thrombin to
produce a sealant or an adhesive.
The apparatus of the invention may be used for other automated
processes. For example, another technique for the separation of
fibrinogen from blood in accordance with the structure of the
invention uses cryoprecipitation. According to this technique,
plasma is frozen to a temperature of about minus 20.degree. C.,
thawed, and then centrifuged to separate the fibrinogen from
plasma. The multiple-decanting apparatus of this invention may be
used to automate cryoprecipitation by inclusion of a temperature
control device 50 in thermal contact with the centrifuge. The
temperature control device may comprise any of several known
structures, including liquid nitrogen or liquid oxygen based
devices and refrigeration devices.
To effect automated cryoprecipitation, a sample of blood is placed
in the first chamber 8, and the container is then placed in the
centrifuge and subjected to a first centrifugation. The plasma is
then drained into the second chamber 8, for example by gravity
draining. The temperature control device is then activated first to
freeze the plasma and then to allow the plasma to thaw. The thawed
plasma is subjected to a second centrifugation, which separates
fibrinogen from the remainder of the plasma. The supernatant plasma
is then separated from the fibrinogen by draining it back into the
first chamber, for example by centrifugal draining, whereby only
fibrinogen remains in the second chamber. The container is then
removed from the centrifuge, and the fibrinogen removed from it for
use as described above. Of course, the freeze-thaw-centrifuge
process may be carded out any number of times before the
supernatant is drained back into the first chamber.
Modifications within the scope of the appended claims will be
apparent to those of skill in the art.
* * * * *