Abstract:
An apparatus for substantially horizontally oscillating one or more vessels containing a liquid, a solid, or a mixture thereof that operates to thoroughly disrupt and mix solid and liquid substances in vessels, like test tubes. The apparatus provides superior mixing through oscillation of the vessels both horizontal and vertical directions through the use of one or more springs and rotating mechanical components. The apparatus includes a circular rotating ring having a toothed circumference that rotates horizontally, at least one vessel support having a compartment configured to hold the one or more vessel vessels and a spring attached to the housing and in constant contact with each of the one or more vessel supports.

Description:
BACKGROUND 
     The present invention describes an apparatus and method for mixing and disintegrating materials in test tubes, particularly for those materials that may be difficult to disintegrate, such as tissues. Devices for these purposes are described in U.S. Pat. Nos. 3,819,158; 4,202,634; 4,295,613; 4,883,644; 4,118,801; 4,125,335; 4,305,668; 4,555,183; 4,747,693, 5,708,861, and 5,769,538. 
     The most common test tube disrupters use tube vibration technology. Tube vibration technology involves a vibrating surface against which test tubes are held by the operator. Vibration of the surface induces vibration of the contents of the tubes. Tube vibration disrupters are simple devices that have several drawbacks. They provide low power and are only effective for disruption of cells and tissues of low hardness. Additionally, they require that the operator have their hand in physical contact with the test tubes, thus subjecting them to the same physical vibrations, which may cause discomfort and increases the potential for receiving injury. 
     U.S. Pat. No. 5,769,538 discloses a more advanced tube striking technology to produce vibrations. A popular brand of disruptor or BULLET BLENDER® uses tube striking technology. The advantages of tube striking technology over tube vibration are that multiple tubes may be processed at once, and the operator need not remain in physical contact with the tubes during disruption. However, these striking-style disrupters have several drawbacks. 
     The transfer of the energy of the strike to the contents of the tube may be inefficient, being dissipated by the liquid media within the tube. This causes an increase in power usage and a decrease in effectiveness. This drawback is particularly disadvantageous when using larger test tubes or when disrupting harder tissues. 
     Strong periodic strikes cause significant vibration, which necessitates the use of vibration dampeners. This is particularly important for hard tissues, such as heart or kidney, which require strong strikes to adequately disrupt. These dampeners increase the weight and cost of the disrupters and increase the rigidity of connections between the parts. These strong strikes also increase the chance of destroying the test tube. 
     Tube strikers are extremely loud, and require expensive, large housings with inner sound isolation to dampen noise. 
     All of the above drawbacks limit the number of potential applications and increase the structural complexity of the devices and increase their weight, size, and production cost, and are particularly disadvantageous for disruption of larger test tubes and harder tissues. 
     SUMMARY 
     The present invention overcomes many of these drawbacks, and allows the user to disrupt small or large sample sizes and to process many samples simultaneously without cross-contamination. The invention provides enhanced disruption and mixing of cells and tissues in test tubes, in the presence of liquid substance and beads, by using a spring mechanism. The core innovation is the interacting of spring mechanisms and specific test tube supports to provide horizontal, vertical, or both horizontal and vertical oscillations of the test tubes. The invention combines the two major functions of oscillations and striking. 
     The tube support includes a body having a compartment that can accommodate a test tube. The spring mechanism attached to the test tube holder pushes the test tube holder horizontally, vertically, or both, depending on which type of oscillation (horizontal or vertical) is intended. 
     For horizontal oscillation, the mechanism includes a rotating ring having a number of curved steps along its inner edge, and springs which hold the test tube supports in constant contact with the inner stepped edge of the rotating ring. The curved steps of the rotating ring may be smooth (like a sine wave) or may have sharp edges. Due to the shape of the rotating ring edge, the test tube support receives horizontal oscillation, which is transferred to the contents of the test tube, which in turn generates chaotic movement and clashing of the beads with the cells and tissues. The more frequent and strong the clashes, the more effective the disruption process. In another embodiment, the curved steps are located on the outer edge of the rotating ring and test tubes supports are located around its periphery. 
     For vertical oscillation, the mechanism includes a rotating disk which has wedges, and springs which hold the test tube supports in contact with the rotating disk. The wedges on the disk are created in similar arrangement to those of the rotating ring, and may also be smooth or sharp-edged, and which are arranged on the disk in a circular pattern about the axis. Due to the shape of the rotating disk, the test tubes receive vertical oscillations, which are transferred to the contents of the test tube, which in turn generates chaotic movement and clashing of the beads with the cells and tissues. The more frequent and strong the clashes, the more effective the disruption process. 
     Horizontal and vertical mechanisms as described above may be used individual within a device, or in a device that includes both mechanisms. Some embodiments may include removable or replaceable rotating disks or rings, allowing the user to configure the type of oscillation to their intended purpose. 
     Embodiments of the invention permit use of a variety of test tube sizes and dimensions. 
    
    
     
       BRIEF DESCRIPTION OF DRAWINGS 
         FIG. 1  shows a vertical cross-sectional view of an embodiment for horizontal oscillation intended for standard small volume 1.5-2.0 ml test tubes. 
         FIG. 2  shows a top section view of an embodiment for horizontal oscillation with a smooth, wave-like rotating ring and one small volume test tube in each of eight tube supports. 
         FIG. 3  shows a top section view of an embodiment for horizontal oscillation with a jagged, rotating ring and one small test tube in each of eight supports. 
         FIG. 4  shows a top section view of an embodiment with a jagged rotating ring and four tube supports, each supporting two test tubes. 
         FIG. 5  shows a vertical cross-sectional view of an embodiment for both horizontal and vertical oscillation for disruption of large volume 50 ml test tubes. 
         FIG. 6  shows a top view of an embodiment&#39;s rotating disk, supporting vertically-extended wedges. 
         FIG. 7  shows a side view of one section of the embodiment in  FIG. 5 . The section illustrates the vertical pulsing motion of the test tubes due to contact with wedges when the rotating disk is rotated. 
         FIG. 8  shows a top view of an embodiment for horizontal oscillation in which the rotating ring is located centrally and test tubes are placed around the periphery. 
         FIG. 9  shows a vertical cross-sectional view of an embodiment for vertical oscillation with a rotating disk and top-mounted springs. 
         FIG. 10A  shows a top view of the rotating disk in  FIG. 9 , with vertically-extending wedges.  FIG. 10B  shows a side view of the wedges on the rotating disk. 
         FIG. 11  shows a vertical cross-sectional view of an embodiment in which test tubes are positioned horizontally rather than vertically. 
         FIG. 12  shows a top section view of the arrangement of test tubes in  FIG. 11 . 
     
    
    
     DEFINITIONS 
     Rotating Ring: A ring with either a curved or jagged inner or outer edge that is rotated laterally on a central axis. 
     Rotating Disk: A disk on which vertical wedges are configured in a circular shape around the axis. 
     DETAILED DESCRIPTION 
       FIGS. 1-12  show embodiments of a spring mechanism used for disruption of cells and tissues in test tubes. 
       FIG. 1  is a side view of an embodiment wherein standard small test tubes  2  are placed in test tube supports  9  which are attached to centrally-mounted stationary springs  6 . The springs  6  keep the test tube supports in constant contact with the inner edge of the rotating ring  8 . Cell or tissue samples  17  along with beads are placed with a liquid buffer  15  in the tubes  2 . The tubes  2  are displaced with respect to the central axis as the rotating ring rotates  8 , causing the liquid buffer  15  clash with the beads. When the liquid buffer  15  and the beads clash, the tissue breaks up and disperses within the test tubes  2 . The stronger and more frequent the clashes, the more effective the disruption and dispersion. 
     The device comprises a housing  1  with a top plate  4  and a base  3 . An electric motor  10  drives a shaft  11  which rotates a bowl  7 . A rotating ring  8  is integral to the inner side of the upper edges of the bowl  7 . Tube supports  9  and compartments  13  are positioned radially about the motor shaft  11 , inside and in constant contact with the rotating ring  8  as it rotates. There is also no gap between the tube support  13  and test tube  2 . 
     In the embodiment shown in  FIG. 1 , flat L-shaped springs  6  are attached with screws to a top plate  4 . The L-shaped springs  6  are also attached with screws to the tube supports  9 . The springs  6  keep the tube supports  9  and tubes  2  in their suspended position, pressed against the rotating ring&#39;s inner edge. 
     Test tubes  2  may be loaded into their tube supports  9  manually. Therefore, the top plate  4  includes holes  5  whose diameter is larger than the diameter of the tube caps  145 . Tubes  2  are inserted into their supports  9  though the holes  5 . 
     The housing  1  has a removable cover  12  which is closed during operation. The main function of the closure is to prevent the tubes  2  from being ejected from their supports  9  while the supports are moving due to contact with the rotating ring  8 . The gap between the tube caps  145  and the closure  12  should be minimal; in some embodiments a spring exerts vertical pressure on the tube cap to keep the tube in place. A sound damping gasket  14  is placed between the closure  12  and the top plate  4  along the outer perimeter. The gasket may be loose and secured by pressure or adhered to either the closure  12  or the top plate  4 . 
     Also shown on  FIG. 1  are feet  100  which support the entire structure. The feet are made of metal but could be made of rubber, or plastic or any material common in the art. The drum  7  and rotating ring  8  are rotated radially by axle  11  which is rotated by an electrical motor  10 . As shown in  FIG. 2 , when rotating ring  8  is rotated counter-clockwise, each tube support  9 , and its respective tube and contents, move laterally. Each support  9  is pressed against the ring&#39;s teeth  20 . The ring&#39;s toothed surface  20  travels along the outer edge of the supports  9 . This radial displacement and force inwardly on the tube supports  9  moves the tube&#39;s  2  beads and liquid buffer  15 , which clash with one another inside the tubes  2 . 
     For example, with eight tubes  2  agitated by eight teeth in the ring  8 , rotating at 600 rpm, the tubes will be oscillated 80 times per second. Vibrations cause the beads to clash with each other and with the cells and tissues in between. These clashes cause disruption of the cells and tissues inside of the test tubes  2 . Varying the speed of rotation of rotating ring  8 , the size of teeth, and the distance between teeth, will cause corresponding changes to the amount and magnitude of the oscillation. 
     For some types of tissues, the disruption process must be accelerated to prevent long-term overheating. This acceleration is accomplished by sharpening the shape of the ring&#39;s teeth  40  ( FIG. 3 ). The angular velocity is labeled as “w” in  FIG. 3 . In this embodiment, each tooth&#39;s recess point  18  and end protrusion point  19  is connected along the shortest radial line  43 , forming a step between adjacent teeth. Due to this drastic step, the tube contents (liquid, cells/tissues and beads) jump or move suddenly with increased acceleration. These jumps release additional energy applied to beads, which intensify and increase their number of clashes within the test tube  2  with each other and the test tube contents. 
     To reduce noise of clashes between tube supports  9  and rotating ring teeth  8 , a rubberized or otherwise sound-dampening layer can be applied to the ring  8  or to the tube supports  9 . 
     Another embodiment shown in  FIG. 4  shows a doubled tube capacity, holding eight tubes, radially paired. This embodiment has tube supports  91  which each house two tubes. This embodiment works in substantially the same way as the previously described embodiment, and it is more compact and efficient than other disrupters in the industry. 
     A third embodiment shown in  FIGS. 5-8  is capable of simultaneously processing big standard tubes of 50 ml, containing tissues of any hardness. The embodiment of  FIG. 5  processes the tissues along two perpendicular axes. In  FIG. 5 , spring  36  extends from top plate  34  to the bottom plate  33  of housing  1 . Spring  36  keep the tube supports  39  touching the revolving ring  38 . Springs  36  are located adjacent to each tube support  39  and are hidden for clarity in  FIG. 5 . Tube supports  39  hold the tubes  32 . The support&#39;s  39  height is approximately ⅓ of its tube&#39;s  32  length. The support is positioned toward the bottom of the tube  32 . The bottom end of tube  32  is supported by rotating disk  310  which is rotated together with the rotating ring  38  by a motor  35 . The support  39  is long enough to prevent wobbling of tube  32  inside the tube support  39  when the apparatus is rotating. Integrally attached to the rotating disk  310  are wedges  314 , which are configured in circles about the axis of the rotating disk. The shape of wedges  314  vary much the same way that teeth  40  vary; the teeth may form a steady sine wave, may resemble jagged teeth, or may be virtually any shape in between. 
     Wedges  314 , which are attached to the rotating disk  310 , provide vertical lift to tubes. The rotating disk  310  rotates together with the ring  38 . Wedges  314  extend upward from the rotating disk  310 ; the wedges are integrated onto the rotating disk. When the rotating disk  310  rotates, wedges  314  periodically travel under the tubes&#39; bottoms and lift up the tubes, adding to the tube&#39;s oscillation. 
     To prevent tubes  32  from being ejected vertically out of their supports  39  when the tubes are pushed up by wedges  314 , springs  336  press down on tube caps  321  as shown in  FIG. 5 . These springs  336  also enhance the energy of internal bead clashes because they produce an opposite downward force on the tubes  32  equal to the input force from the rotating disk. The beads inside the solution react with the contents of the tube as the tube is moved down by the spring or up rotating disk, causing more complete mixing and disruption of the substance  351 . Springs  336  and  337  are attached to the top cover via traditional fastening means, and lay freely against the tube cap. F 1  and F 2  represent the varying forces applied by the springs  336  and  337 . The force varies depending on whether a wedge is causing vertical displacement on the tube. 
     For larger tube processing, which can generate unwanted heat, one or more fans  41  powered by a motor  35  can blow air toward the tubes. A dry air box may also be placed beneath the housing knot shown). Both the one or more fans  41  and the dry air box may be a means for cooling the motor  35  and one or more vessels. 
     A fourth embodiment is shown in  FIGS. 9 and 10 .  FIG. 9  shows a vertical cross-section of a device for processing small tubes by only vertical pulsation of the tubes, similar to the method for vertical disruption described above. In this embodiment, no horizontal displacement occurs. 
     In  FIG. 9 , tubes  104  are shown moving vertically caused by the springs  107  providing forces F 1  and F 2  to react the vertical upward force created when the bottom end of the tubes  101  and  104  interface with a wedge  110  on the rotating disk  109 . The springs  107  are attached to a cover  102  using bolts or other standard fastening means. The rotating disk  109  is powered by a motor  112 . The tube is supported by a tube support  106  which is integral to the housing  105  which has openings into which the tubes are placed  106 . 
       FIG. 10B  shows a side view of the wedges  110  on the rotating disk  109 . The profile and shape of the wedges can take many different forms from smooth curves to sharp steps as shown in  FIG. 10B . Also shown in  FIG. 10A  is a top view of the rotating disk which shows the location of the various tube bottoms  104  and the placement of the wedges  110  on the rotating disk  109 . 
     A fifth embodiment is shown in  FIGS. 11-12 , in which test tubes supports  203 , compartments  205 , and test tubes  200  are arranged horizontally rather than vertically. Tube supports  203  and compartments  205  are built to accommodate tubes in a horizontal position. The rotating ring  204  is constructed substantially the same way as in the other embodiments. To prevent test tubes  200  from being ejected, top cover  202  has extensions  201  made of a material with a low friction factor with the material of the tubes.