The present disclosure relates to a multiple component sample concentrator/separator. More particularly, the present disclosure relates to a multi-lobed centrifuge configured to separate and concentrate various biological components.
Bone marrow aspiration involves inserting a needle into bone and withdrawing a material from the bone. The withdrawn material, for instance withdrawn bone marrow aspirate or “BMA,” can contain multiple components including plasma, red blood cells, and a buffy coat layer (that includes stem cells). After withdrawal of the multiple component sample the multiple components are often mixed together such that collection of a concentrated sample of any single component can be difficult. The multiple component sample can be separated into various components including, for instance a desired component (such as the buffy coat) and a remaining component (such as the plasma and red blood cells).
One process that can be used to separate the desired component from the remaining component of the multiple component sample is centrifugation. During centrifugation of the multiple component sample, for instance within a centrifuge device, each of the multiple components in the sample will assume a particular radial position within the device based upon the respective densities of each of the components. The multiple components will therefore separate when the centrifuge device is rotated at an appropriate angular velocity for an appropriate period of time.
Referring to
Due to the intermediate position of the buffy coat 5 between the red blood cells 3 and the plasma 7 and also due to the relatively small size of the buffy coat layer 5 relative to the red blood cell layer 3 and the plasma layer 7, extraction of a concentrated volume of the buffy coat 5 after centrifugation can be difficult. One means of eliminating the red blood cell layer 3 is by lysing the red blood cells. A centrifugation device that enables the recovery of a high percentage of the desired component at a high concentration could result in time and cost savings for certain procedures.
The present disclosure provides, in accordance with one embodiment, a collection tray configured to rotate about an axis of rotation to separate a multiple component sample into a desired component and a remaining component. The collection tray can include a ray line that extends perpendicularly from the axis of rotation. The collection tray can include a collection body configured to receive the multiple component sample, and a plurality of lobes supported by the collection body. Each of the lobes can have two lobe base portions, an apex, and two lobe side walls that each extend between one of the lobe base portions and the apex. At least one of the lobes can define a straight lobe line that perpendicularly intersects one of the lobe side walls at a point located radially between the respective lobe base portion and the apex, such that the ray line intersects the point so as to define a lobe angle measured between the ray line and the lobe line. The lobe angle of the collection tray is greater than a specific angle, such that the arctangent of the specific angle is equal to the effective coefficient of friction of the desired component and the lobe side wall.
In accordance with another embodiment, the present disclosure provides a device configured to separate a multiple component sample into a desired component and a remaining component. The device includes a bowl portion defining an interior configured to receive the multiple component sample, and the bowl portion is configured to rotate about an axis of rotation. The device further includes a collection tray configured to be supported by the bowl portion so as to rotate about the axis of rotation. The collection tray defines a ray line that extends perpendicularly from the axis of rotation, and the collection tray includes at least one lobe that has two lobe base portions, an apex, and two lobe side walls that each extend from one of the lobe base portions to the apex. The at least one lobe at least partially defines a basin that is in fluid communication with the interior of the bowl portion such that the multiple component sample is transferable from the interior to the basin during rotation of the bowl portion about the axis of rotation. The at least one lobe further defines a lobe line that is different from the ray line, and the ray line intersects one of the lobe side walls at a point along the lobe side wall. The lobe line perpendicularly intersects the point so as to define a lobe angle between the ray line and the lobe line.
In accordance with another embodiment, the present disclosure provides a process to process a withdrawn BMA sample. The process includes the steps of: combining the withdrawn BMA sample and a red blood cell lysing agent so as to form a multiple component sample; rotating a device about an axis of rotation, the device containing the multiple component sample, so as to separate the multiple component sample into a desired component and a remaining component; and collecting at least a portion of the desired component.
The foregoing summary, as well as the following detailed description of the preferred embodiments of the application, will be better understood when read in conjunction with the appended drawings. For the purposes of illustrating the surgical instruments and methods of the present application, there is shown in the drawings preferred embodiments. It should be understood, however, that the application is not limited to the specific embodiments and methods disclosed, and reference is made to the claims for that purpose. In the drawings:
Certain terminology is used in the following description for convenience only and is not limiting. The words “upper”, “lower”, “above” and “below” designate directions in the drawings to which reference is made. The terminology includes the above-listed words, derivatives thereof and words of similar import. Additionally, a radial or polar coordinate system is provided and described herein. The polar coordinate system includes a two dimensional radial plane that is centered on and normal to an axis, for instance an axis of rotation. The polar coordinate system defines a radial component that is measured as the distance from the axis along the plane. The words “inner” and “outer” designate locations closer to and farther away from the axis respectively. The polar coordinate system further defines an angular component that is measured as the angular position about the axis. The radial coordinate system can be converted to a three dimensional coordinate system, for instance a right-hand coordinate system that includes a first or longitudinal direction L, a second or lateral direction A that is perpendicular to the longitudinal direction L, and a third or transverse direction T that is perpendicular to both the longitudinal direction L and the lateral direction A. The longitudinal direction L and the lateral direction A can define a plane that corresponds to the radial plane and position along the radial axis corresponds to position in the transverse direction T.
The term “plurality”, as used herein, means more than one. When a range of values is expressed, another embodiment includes from the one particular value and/or to the other particular value. Similarly, when values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another embodiment. Further, reference to values stated in ranges includes each and every value within that range. All ranges are inclusive and combinable. Certain features of the invention which are described herein in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention that are described in the context of a single embodiment, may also be provided separately or in any subcombination.
Referring to
Referring to
As will be described in greater detail below, the device 18 can be configured such that the desired component, for example the cell pellet 15, of the lysed and centrifuged BMA 1, has a volumetric concentration of stem cells, that is greater than the average volumetric concentration of stem cells of the withdrawn BMA 1 prior to lysing and centrifugation. In accordance with one embodiment, the volumetric concentration of stem cells in the cell pellet 15 can be at least a multiple, such as four fold, of the average volumetric concentration of stem cells in the withdrawn BMA 1. In one embodiment, the device 18 can be configured such that the cells of the desired component, for instance the stem cells of the cell pellet 15, maintain at least 95% viability during separation and collection by the device 18. In one embodiment, the device 18 is configured to complete the separation and collection of the desired component from the remaining component in 30 minutes or less, such that the device 18 can be used intraoperatively.
The device 18 can be configured to accept a range of volumes of withdrawn BMA 1. The volume of withdrawn BMA 1 can be separated into the desired component and the remaining component and the desired component can then be collected. In one embodiment, the device 18 is configured to accept and separate any volume of withdrawn BMA 1 as desired, for example between about 8 cc to about 50 cc. Furthermore, it should be appreciated that the withdrawn BMA 1 can be lysed either prior to introduction into the device 18 or after introduction to the device 18. In one embodiment, the device 18 is configured to be ergonomic and intuitive such that the device 18 is easy to use in an operating room environment. The device 18 can be configured such that the separator 20, collector 100, and the housing 300 can be double packaged and sterilized. The device 18 can also be configured to be disposable, such that after the separation and collection of the desired component of a multiple component sample, the device 18 can be thrown away. The device 18 can further be configured to provide maximum portability such that the device 18 is cordless or self-contained, for instance battery powered with no external power source needed.
Referring to
The bowl body 32 includes a bowl bottom 34, an upper lip 36, and a height H1 measured from the bowl bottom 34 to the upper lip 36. The bowl body 32 further includes a bowl wall 38 that extends from the bowl bottom 34 to the upper lip 36 and an inner diameter D1 that is measured from one side of the bowl wall 38 to another side of the bowl wall 38 along a straight line that passes perpendicularly through the axis of rotation 22. The bowl wall 38 is angularly offset from the axis of rotation 22 such that the inner diameter D1 of the bowl body 32 gradually increases from a minimum value at the bowl bottom 34 to a maximum value at the upper lip 36. The bowl body 32 can be configured with a height H1 and an inner diameter D1 such that the bowl portion 24 can be filled with a range of volumes of multiple component sample and still produce effective separation of the desired component from the remaining component.
In one embodiment the height H1 and the inner diameter D1 of the bowl body 32 are configured such that the bowl portion 24 is capable of receiving any desired volume of withdrawn BMA 1, such as between about 8 cc to about 50 cc in addition to a volume of lysing agent. The amount of lysing agent can be, for example, twice the volume of the volume of withdrawn BMA 1. Thus for a volume of withdrawn BMA 1 between about 8 cc to about 50 cc, the volume of lysing agent can be between about 16 cc to about 100 cc. Thus, the dimensions of the bowl body 32, including the height H1 and the inner diameter D1 can be chosen from a range of values such that the bowl portion 24 is configured to receive a range of total volume of withdrawn BMA 1 and lysing agent between about 24 cc to about 150 cc.
Referring to
Each of the lobes 46 includes two base portions 48 and an apex 50 disposed radially farther from the axis of rotation 22 than each of the two base portions 48. In another embodiment, the lobes 46 can include more than two base portions 48. Each of the lobes 46 further includes two lobe side walls 52 that each extends between one of the two base portions 48 and the apex 50. In one embodiment, the lobe side wall 52 is the radially outward most component of the collection body 44. Each of the lobe side walls 52 can include an inner side wall 53 and an outer side wall 55, the outer side wall 55 being disposed radially farther from the axis of rotation 22 than the inner side wall 53. In one embodiment, the outer side wall 55 is the radially outward most component of the collection body 44. Each of the lobes 46 can further include a floor 51 that extends at least partially radially in a first direction between the inner rim 40 and the apex 50 and angularly in another direction between the inner side wall 53 of each of the side walls 52 of the respective lobe 46. The inner side walls 53 of the two side walls 52 and the floor 51 together define a basin 57 of the lobe 46 that is configured to receive a volume of the multiple component sample during rotation of the separator 20 about the axis of rotation 22.
The lobes 46 can be configured such that the cumulative volume of all of the basins 57 of all of the lobes 46 is greater than or equal to the total volume of the desired component that will be separated from the remaining component after centrifugation of the multiple component sample. For example if a 50 cc sample of withdrawn BMA 1 is placed in the separator 20 along with a 100 cc sample of lysing agent, the desired component is a fraction, for instance one-twelfth or 12.5 cc of the total volume of lysed BMA 11. In this example, if the separator 20 includes a collection tray 26 with four lobes 46, each of the basins 57 of the four lobes 46 could be configured to define a volume of at least 3.2 cc. A variety of collection trays 26 can include a number of different configurations of lobes 46 with basins 57 that define various volumes to accommodate samples of various volumes and desired ratios of total sample to desired component.
In one embodiment, the side walls 52 define a midpoint 59 that is located radially halfway between the base portion 48 and the apex 50. The side walls 52 each include a proximal portion 61 located between the base portion 48 and the midpoint 59 and a distal portion 63 located between the midpoint 59 and the apex 50. The side walls 52 can be curved, for instance such that the inner side wall 53 is concave and the outer side wall 55 is convex, as shown in the illustrated embodiment. In one embodiment, the inner side wall 53 within the proximal portion 61 of lobe 46 is curved such that no portion of the inner side wall 53 is parallel to a radial ray 65 that extends straight out from the axis of rotation 22 and intersects the apex 50 of the respective lobe 46. In another embodiment, the inner side wall 53 is curved such that no portion of the inner side wall 53 extends purely radially (or only in the radial direction).
The lobes 46 each define a lobe angle β that is measured between a radial ray 54 (a straight line extending from and perpendicular to the axis of rotation 22 to a point on the inner side wall 53) and a lobe line 56 (a straight line that perpendicularly intersects the inner side wall 53 at the point). In one embodiment, the collection body 44 of the collection tray 26 is configured such that the lobe angle β has a desired value greater than a certain value (referred to herein as the “specific value”). The specific value of the lobe angle β is defined such that when a component of the sample, such as a mononucleated cell, is in contact with the inner side wall 53 and a radial force is applied to the component of the sample (such as centripetal force when the bowl portion 24 and the collection tray 26 are spinning, or rotating, about the axis of rotation 22), the component of the sample will move relative to the inner side wall 53. In one embodiment, the specific value of the lobe angle β can be determined by calculating the inverse tangent or the arctangent (TAN−1) of the effective coefficient of friction of the desired component and the inner side wall 53. The calculation can be represented by the following equation: (specific value)=TAN−1 (effective coefficient of friction).
For example, referring to
The material of the inner side wall 53, the surface smoothness of the inner side wall 53, and the constituents of the multiple component sample can all affect the effective coefficient of friction and therefore the specific value. In one embodiment a coating, can be applied to inner side wall 53 to change the effective coefficient of friction between the inner side wall 53 and the desired component. In one embodiment, PTFE (Teflon) coating can be applied to the inner side wall 53, for instance by spraying or a mechanical process. Once the specific value for the lobe angle β has been determined the separator 20 can be configured with a lobe angle β that is chosen to be greater than the specific value. The actual lobe angle β can be chosen based on additional factors related to ease of construction and operation, size restrictions, ease of collection, etc.
Referring again to
In one embodiment, the lobe angle β can be substantially constant measured at any point along the side wall 52. As shown in the illustrated embodiment, the lobe angle β can be measured at a first point 52a near the base portion 48, at a second point 52b near the apex 50, or at a third point 52c nearly midway between the base portion 48 and the apex 50. In one embodiment, the lobe angle β is substantially the same at first point 52a, second point 52b, and third point 52c. In another embodiment, the lobe angle β can vary as measured at different points along the side wall 52. For example the lobe angle β measured at each of the first, second, and third points 52a, 52b and 52c, can be different, but always greater than the specific value.
The lobes 46 can further include an inner tray surface 58 and an opposed outer tray surface 60. As shown in the illustrated embodiment, the inner tray surface 58 can define a negative slope such that the inner tray surface 58 extends downward (in a direction from the inner rim 40 toward the bowl bottom 34 and parallel to the axis of rotation 22) and radially outward (in a direction from the inner rim 40 toward the tray outer periphery 42 and perpendicular to the axis of rotation 22).
The inner tray surface 58 defines a collection area, such as a pocket 62 that is configured to collect a concentrated sample of the densest component of the multiple component sample during rotation of the separator 20 about the axis of rotation 22. In one embodiment the pocket 62 is the radially most distant part of the basin 57. The negative slope of the inner tray surface 58 is configured such that when the bowl portion 24 stops rotating about the axis of rotation 22, the densest component of the multiple component sample, for instance the cell pellet 15 in a sample of lysed BMA 11, is retained in the pocket 62 for collection. In one embodiment the inner tray surface 58 defines a vertical offset 64 that is the distance between the pocket 62 and the inner rim 40 as measured along a direction parallel to the axis of rotation 22 (or in the transverse direction T). As shown in the illustrated embodiment, the vertical offset 64 can be configured such that a portion of the basin 57 is located below (or downward relative to) the inner rim 40. In one embodiment, the cumulative volume of the basin 57 of each of the lobes 46 that is located below the inner rim 40 is equal to or greater than the volume of the desired component of the multiple component sample.
Although the collection tray 26 and bowl portion 24 are shown as integral or monolithic parts in the illustrated embodiment, in another embodiment, the collection tray 26 can be a separate or separable part with respect to the body portion 24 such that a collection tray 26 with a desired lobe angle β can be chosen from a kit containing a plurality of collection trays 26 with a plurality of lobe angles β, based on the particular multiple component sample that is to be separated. In this embodiment the collection tray can be a monolithic body such that each of the lobes 46 are integral (or not easily separable) with one another. Once the collection tray 26 with the desired lobe angle β is chosen, the collection tray can be attached to the bowl portion 24.
As shown in
Referring to
Referring to
The lid 70, as shown in the illustrated embodiment can be centered on the axis of rotation 22 such that the lid 70 is configured to spin or rotate about the axis of rotation 22 when the lid 70 is secured to the collection tray 26. The lid 70 defines a lid outer periphery 72, and the lid 70 includes a lid body 74 that extends radially between the axis of rotation 22 and the lid outer periphery 72. The lid body 74 can include a lobe portion 76 and a dome portion 78. The lobe portion 76 can include lid lobes 80 that correspond (for example, in number and shape) to the lobes 46 of the collection body 44. The lobe portion 76 can further include a lid inner surface 81 that along with the inner tray surface 58 defines the pocket 62 when the lid body 74 is properly secured to the collection body 44.
Referring to
Referring to
In one embodiment, for instance as shown in
In another embodiment, the lid body 74 may be secured to the collection body 44 using an adhesive, which may also fill any potential gaps between the lid body 74 and the collection body 44.
Referring to
As the bowl portion 24 begins to rotate about the axis of rotation 22, as shown in
After substantially all of the cell pellet 15 has been separated from the supernatant 13 and concentrated in the pocket 62, rotation of the bowl portion 24 and the lid 70 about the axis of rotation 22 can be terminated. As shown in
Referring to
The collector 100, as shown in the illustrated embodiment, can further include a collection container, for instance a syringe 118, that is configured to be supported by the housing 104, for example at an attachment point 125, and collect and contain an amount of a concentrated sample of a desired component of a multiple component sample. The collection container is connected to the free end 108 of the probe 102 such that the desired component collected by the probe 102 is transferred to the collection container. For example, the syringe 118 can be pneumatically connected to the free end 108 of the probe 102.
The collector 100 can further include a guide rod 112 with a first end 114 and a second end 116 opposite the first end 114. In one embodiment, the housing 104 is configured to translate along the guide rod 112 from a first contracted configuration (as shown in
The collector 100 can additionally include one or more scrapers 120, that are configured to aid in the collection a concentrated sample of a desired component of a multiple component sample. Each of the one or more scrapers 120 can be attached to the housing 104, for instance to a flange 121 of the housing 104 such that the probe 102, the housing 104, and the at least one scraper 120 are all translationally locked relative to each other, such that as the housing translates along the guide rod 112, for example in the radial or specifically in the longitudinal direction L, the probe 102 and the at least one scraper 120 also translate along with the housing 104 in the same direction as the housing 104. As shown in the illustrated embodiment, the collector 100 can include a body 103 that functions both as the scraper 120 and as the probe 102 are described within the present disclosure.
The collector 100 defines a passage 122 from the free end 108 of the probe 102 to the attachment point 125 of the syringe 118. The passage 122 provides a path for the collected sample to pass through the collector 100 from the free end 108 of the probe 102 to a receiving chamber 119 of the syringe 118. As shown in the illustrated embodiment, the probe 102 defines a cannula 110 (shown in dashed lines) that extends through the body 105 from the free end 108 to the attached end 106. The collector 100 can further include a tube 123 that is connected, for example pneumatically, to the attached end of the probe 102. In one embodiment the tube 123 at least partially defines the passage 122, and is pneumatically connected to the attachment point 125.
Once the collector 100 has been moved into the second expanded configuration such that the free end 108 of the probe 102 is positioned within the desired component of the multiple component sample, a plunger 128 of the syringe 118 can be actuated to draw the desired component into the passage 122 for collection. As the plunger 128 is actuated, the desired component adjacent the free end 108 of the probe is drawn into the cannula 110 of the probe 102 at the free end 108. The desired component is then drawn in a direction toward the attached end 106 of the probe 102 (or proximally). The desired component is next drawn into the tube 123 which connects the probe 102 to the syringe 118. It will be apparent to one of skill in the art that the arrangement and selection of the components of the collector 100 that form the passage 122, for example the tube 123, could be changed or substituted without deviating from the teachings of the present disclosure.
Referring to
Referring to
In one embodiment the bracket 130 includes an inner bore that is configured to receive the guide rod 112. Once the guide rod 112 has been received within bracket 130, the guide rod 112 can be secured relative to the bracket 130 such that the guide rod 112 and the bracket 130 do not move relative to one another, for instance by a friction fit between the guide rod 112 and the inner bore of the bracket 130. In another embodiment the bracket 130 can include a set screw or other fastener configured to be received within a recess of the bracket 130 and tightened against the guide rod 112 to secure the guide rod 112 relative to the bracket 130. The housing 104 and probe 102 can translate along the guide rod 112 from the first contracted configuration (as shown in
As described in detail above, separation of a multiple component sample (lysed BMA 11) into its separate components (cell pellet 15 and supernatant 13) can be performed by rotation of the bowl portion 24 and the lid 70 about the axis of rotation 22. As shown, the desired component (cell pellet 15) is concentrated within the pocket 62 near the apex 50 of the lobes 46 of the collection body 44. The collector 100 can then be moved into the second expanded configuration such that the free end 108 of the probe 102 is positioned within the desired component, for instance cell pellet 15, of the multiple component sample. The collector 100 can then collect a sample of the cell pellet 15 or other desired component.
In one embodiment the bracket 130 can be secured to the housing 300 such that bracket 130 and the secured collector 100 are in a fixed radial position relative to the separator 20. Thus to collect the desired component from a first of the lobes 46, the collection body 44 can be rotated about the axis of rotation 22 until a reference point of the collector 100, for example the guide rod 112 is aligned with the apex 50 of one of the lobes 46. In another embodiment, the reference point can be the free end 108 of the probe 102. The collector 100 can then be transitioned from the first retracted configuration to the second expanded configuration enabling the probe 102 to collect a sample of the desired component of the multiple component sample. During the transition from the first retracted configuration to the second expanded configuration the housing 104 translates along the guide rod 112 in a direction toward the apex 50 of the lobe 46 with which the collector 100 has been aligned. The housing 104 is translated until the free end 108, and the cannulation 110, of the probe 102 is positioned within the cell pellet 15.
The collector 100 can then be actuated, for example by moving the plunger 128 to create a negative pressure within the cannulation 110, which is pneumatically connected to the syringe 118. The negative pressure within the cannulation 110 draws the cell pellet 15 into the free end 108 of the probe 102 and through the passage 122 until the cell pellet is deposited within the receiving chamber 119 of the syringe 118. Once the desired component has been collected from the lobe 46, the collector 100 is transitioned back into the first retracted configuration. Then the collection body 44 can be rotated again until the probe 102 is aligned with the apex 50 of another lobe 46. The process described above can then be repeated until the desired component has been collected from each of the lobes 46.
Referring to
The moveable bracket 130 relative to the lid 70 enables a reference point of the collector 100, for example the guide rod 112 or the probe 102, to be aligned with the apex 50 of one of the lobes 46. The collector 100 can be transitioned from the first retracted configuration into the second expanded configuration such that the free end 108 of the probe 102 is disposed within the desired component. After a concentrated sample of the desired component has been collected (as described above) the collector 100 can be transitioned back into the first retracted configuration. The bracket 130 and the attached collector 100 can then translate along the lip 94 of the opening 82 such that the collector 100 rotates about the axis of rotation 22 relative to the lid 70 and the bowl portion 24 until the collector 100 is aligned with the apex 50 of another lobe 46. The process can then be repeated until a concentrated sample of the desired component has been collected from each lobe 46.
Referring to
Referring to
According to one embodiment, the collector 100 includes a housing 104 that is movably attached to a guide rod 112. The collector 100 further includes a probe 102 that is supported by the housing 104, such that the probe 102 is configured to collect the cell pellet 15. The collector 100 can further include a scraper 120 that is supported by the housing 104. In one embodiment, the probe 102 and the scraper 120 are each attached on opposite sides of the housing 104, for example the probe 102 and the scraper 120 can be attached to the flanges 121 of the housing 104. The probe 102 and the scraper 120 are each secured relative to the housing 104 such that the probe 102 and the scraper 120 each translate along with the housing 104 as the collector 100 is transitioned from the first contracted configuration to the second expanded configuration.
As shown in the illustrated embodiment, the scraper 120 includes an attached end 134 that can be secured to the housing 104, a free end 136 opposite the attached end 134, and a scraper body 138 that extends from the attached end 134 to the free end 136 along a central scraper axis 140. In one embodiment, the central scraper axis 140 can be curved as shown. In another embodiment, the central scraper axis 140 can be substantially straight. Similarly, the probe 102, in one embodiment can extend from the attached end 106 to the free end 108 along a central probe axis 141. In one embodiment, the central probe axis 141 can be curved as shown. In another embodiment, the central probe axis 141 can be substantially straight. In another embodiment, the collector 100 can include the probe 102 that is configured to collect substantially the entire cell pellet 15 without the inclusion of the scraper 120.
In use, the collector 100 is configured to be aligned with one of the lobes 46, for example such that the guide rod 112 is aligned with the apex 50. Once the collector 100 is aligned with the lobe 46 the collector 100 is transitioned from the first retracted configuration to the second expanded configuration, the probe 102 translates with the housing 104 along the guide rod 112 in a direction, for example radially or specifically in the longitudinal direction L, toward the apex 50. As the housing 104 and the probe 102 translate in the radial direction toward the apex 50, the free end 108 of the probe 102 can, in one embodiment, be advanced into the lobe 46 until the free end 108 is positioned within the supernatant 13. The collector 100 can then be actuated, as described in greater detail below, to remove a portion, for example substantially all, of the supernatant 13 from the lobe 46. In one embodiment, the removal of the supernatant 13 can be repeated for all of the lobes 46 of the collection tray 26.
The free end 108 of the probe 102 can then be advanced further in the radial direction until the free end 108 is positioned within the pocket 62 and within the cell pellet 15. The collector 100 can then be actuated, as described in greater detail below, to remove a portion, for example substantially the entirety of the cell pellet 15 from the lobe 46. In one embodiment, the removal of the cell pellet 15 can be repeated for all of the lobes 46 of the collection tray 26. In another embodiment, the free end of the probe 102 can be advanced through the supernatant 13 and into the cell pellet 15 without withdrawing the supernatant 13.
Referring to
In one embodiment, the collector 100 includes a probe 102, for example the probe/scraper body 103, and a scraper 120 that are each supported by the housing 104 of the collector 100. In one embodiment, the probe 102 and the scraper 120 are each attached on opposite sides of the housing 104, for example the probe 102 and the scraper 120 can be attached to the flanges 121 of the housing 104. The probe 102 and the scraper 120 are each secured relative to the housing 104 such that the probe 102 and the scraper 120 each translate along with the housing 104 as the collector 100 is transitioned from the first contracted configuration to the second expanded configuration. As shown in the illustrated embodiment, the scraper 120 includes an attached end 134 that can be secured to the housing 104 as shown, a free end 136 opposite the attached end 134, and a scraper body 138 that extends from the attached end 134 to the free end 136 along a central scraper axis 140.
In one embodiment, the central scraper axis 140 can be curved as shown. Similarly, the probe 102, in one embodiment can extend from the attached end 106 to the free end 108 along a central probe axis 141. The scraper body 138 defines a length measured from the attached end 134 to the free end 136 along the central scraper axis 140. The scraper body 138 can further include a tip portion 142 that is configured to aid in the collection of a concentrated sample of a desired component of a multiple component sample.
In use, as the collector 100 is transitioned from the first retracted configuration to the second expanded configuration, the probe 102 and the scraper 120 each translate with the housing 104 along the guide rod 112 in a direction, for example radially or specifically in the longitudinal direction L, toward the apex 50. As the housing 104, the probe 102 and the scraper 120 translate in the direction toward the apex 50, the tip portion 142 of the scraper 120 and the free end 108 of the probe 102 each move into contact one of the side walls 52 near the base portion 48 (as shown in
As the collector 100 continues to transition from the first retracted configuration to the second expanded configuration, and the probe 102 and the scraper 120 continue to advance in the radial direction toward the apex 50, the tip portion 142 of the scraper 120 and the free end 108 of the probe 102 each translate along the side wall 52 gathering and moving the additional portion of the cell pellet 15′ toward the portion of the cell pellet 15 in the pocket 62 adjacent the apex 50 (as shown in
Once the collector 100 has fully transitioned into the second expanded configuration (as shown in
Referring to
Referring to
The housing 300 includes a top surface 302, a bottom surface 304, and a housing body 306 that extends from the top surface 302 to the bottom surface 304. The housing body 306 can include a base portion 308 and a cap portion 310. The base portion 308 defines an inner cavity 312 that is configured to enclose the separator 20. The separator 20 can be mounted within the inner cavity 312 such that the separator can rotate without interference from the housing body 306. The inner cavity 312 can additionally enclose a motor 400 and a drive shaft 402 rotationally coupled to the motor 400. The drive shaft 402 can be rotationally coupled to the recess 37 of the engagement mechanism 33 of the separator 20 such that the motor 400 can provide a rotational force to the separator 20 that causes the separator to rotate about the axis of rotation 22.
The base portion 308 can further include a window 314 (or other opening) such that an operator of the device 18 can see the separator 20. The window 314 can be configured such that the pocket 62 of the separator is visible through the window 314 allowing for visualization of the pocket 62 during alignment of the pocket 62 with the collector 100 and collection of the desired component from the pocket 62 by the collector 100. Additionally, the device 18 can include a power supply, for example batteries, to power any electrical components of the device 18. The device 18 can further include a printed circuit board that is configured to support and connect electronic components of the device 18 and provide various logic functions. One or more LEDs 320 can be included to indicate the status of the device 18 (e.g., ready to centrifuge, centrifuging, centrifuging complete and ready for collection).
The cap portion 310 is configured to be secured to the base portion 308 to at least partially enclose the separator 20 and the collector 100. During rotation of the separator 20 about the axis of rotation 22, the cap portion 310 can prevent an operator of the device 18 from touching any moving parts of the device 18 during the centrifugation process. In one embodiment, the device 18 includes a cap sensor switch and linkage configured to detect if the cap portion 310 is correctly in place relative to the base portion 308, and allow the motor 400 to spin only if the cap portion 310 is correctly in place relative to the base portion 308. After rotation of the separator 20 has completed and during collection of the desired component, the cap portion 310 can be removed from the base portion 308 such that access to the collector 100 is provided to an operator of the device 18. The housing body 306 can further include a ledge 316 that is positioned between the base portion 308 and the cap portion 310. The ledge 316 is configured to receive the bracket 130 such that the collector 100 is positioned relative to the separator 20 such that when the collector 100 is in the first retracted configuration (as shown in
Referring to
In use, the separator 20 containing the lysed BMA 11 can rotate around the axis of rotation 22 at a desired angular velocity for a desired amount of time, for example 3000 RPMs (or about 500 G's) for about 5 minutes, such that the cell pellet 15 will ride up the bowl wall 38 (due to the bowl angle θ as described above), over the upper lip 36 and into the collection tray 26. As the separator 20 continues to rotate about the axis of rotation 22 the cell pellet 15 will pass into the basin 57 of the lobe 46 and move radially away from the axis of rotation 22 and collect in the pocket 62. The cell pellet 15 can then be collected from the pocket 62 of each of the lobes 46 by the collector 100.
In one embodiment, if a relatively smaller volume of lysed BMA 11 has been centrifuged, the resulting cell pellet 15 may only fill a portion of the pocket 62. The collector 100 can be transitioned to a third intermediate configuration in which the collector 100 is partially transitioned from the first retracted configuration to the second expanded configuration. In the third intermediate configuration the free end 108 of the probe 102 is positioned within the remaining component and close to, but not within the desired component, for example the cell pellet 15. In one embodiment the third intermediate position is determined visually, through the window 314 in the base portion 308. In another embodiment, the collector 100 can include a series of markings 127, for example on the guide rod 112, such that when the housing 104 is aligned with the appropriate marking 127 (based on the initial volume of BMA), the collector 100 is in the third intermediate configuration.
A waste syringe 118 can be connected to the attachment point 125 and the collector 100 can be actuated such that the remaining component is removed from the pocket 62 and drawn into the waste syringe 118. Once the remaining component has been removed from the pocket 62, the waste syringe 118 can be removed from the attachment point 125 and replaced by a second syringe 118. In another embodiment, once the remaining component has been removed from the pocket 62, the collector 100 can be transitioned into the first retracted configuration. The collector 100 can then be aligned with another of the lobes 46 and the remaining steps above repeated until the remaining component has been removed from all of the lobes 46. The waste syringe 118 can be removed from the attachment point 125 and replaced by a second syringe 118.
The collector 100 can then be fully transitioned into the second expanded configuration such that the free end 108 of the probe 102 is disposed within the desire component. The collector 100 can then be actuated to draw the desired component into the second syringe 118 for collection. The collector can then be transitioned back into the first retracted configuration and the second syringe 118 can be removed from the attachment point 125. This process can then be repeated as needed for the remaining lobes 46.
In another embodiment, if a relatively larger volume of lysed BMA 11 has been centrifuged, the resulting cell pellet 15 may substantially fill the pocket 62. In this case, a syringe 118 can be attached to the attachment point 125, the collector 100 can be transitioned from the first retracted configuration to the second expanded configuration, the collector 100 can be actuated to create a negative pressure within the passage 122, drawing the desired component into the probe 102 through the passage 122 and into the syringe 118. The collector 100 can then be transitioned from the second expanded configuration to the first retracted configuration. The collector 100 can then be aligned with the apex 50 of another lobe 46, and the process repeated as needed for any remaining lobes 46.
In one embodiment, the collector can be used to remove at least a portion of the supernatant 13 from the basin 57 of each of the lobes 46. Then the collector can also be transitioned from the first retracted configuration to the second expanded configuration causing the scrapers 120 of the collector 100 to ride along the inner side walls 53 of each of the lobes 46 to gather the cell pellet 15 in each of the pockets 62. The collector 100 is then transitioned from the second expanded configuration to the first retracted configuration before the separator 20 is again rotated about the axis of rotation 22 to concentrate the cell pellet 15 in the pockets 62 at the most radially distant location within the lobes 46. The collector 100 is then again transitioned to the second expanded configuration and a sample of the cell pellet 15 is collected from the pocket 62 of each of the lobes 46. If any cell pellet 15 remains in the lobes 46 the rotation and collection steps can be repeated as desired.
In another embodiment, a solution that loosens the cell pellet 15 from the inner side walls 53 of the lobes can be used between rotation cycles to increase the amount of cell pellet 15 gathered during each collection phase. Once the desired amount of cell pellet 15 has been collected the device 18 can either be disposed of or broken down and sterilized for re-use.
It will be appreciated by those skilled in the art that changes could be made to the embodiments described above without departing from the broad inventive concept thereof. It is understood, therefore, that this disclosure is not limited to the particular embodiments disclosed, but it is intended to cover modifications within the spirit and scope of the present disclosure as defined by the claims.