CA1178183A - Disaggregation device for cell suspensions - Google Patents
Disaggregation device for cell suspensionsInfo
- Publication number
- CA1178183A CA1178183A CA000402150A CA402150A CA1178183A CA 1178183 A CA1178183 A CA 1178183A CA 000402150 A CA000402150 A CA 000402150A CA 402150 A CA402150 A CA 402150A CA 1178183 A CA1178183 A CA 1178183A
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- Canada
- Prior art keywords
- rotor
- beaker
- diameter holes
- cap
- sample suspension
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M45/00—Means for pre-treatment of biological substances
- C12M45/02—Means for pre-treatment of biological substances by mechanical forces; Stirring; Trituration; Comminuting
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- Mechanical Engineering (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Sustainable Development (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Centrifugal Separators (AREA)
- Sampling And Sample Adjustment (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
ABSTRACT
Disclosed is a disaggregating device for separating clusters of biological cells in a sample suspension, comprising: a beaker containing the suspension, a rotor rotatably mounted inside the beaker, a motor for rotating the rotor, the rotor being spaced apart from the walls of the beaker and rotated at a sufficient speed to create shear forces on the clusters of cells located in the sample suspension between the rotor and the beaker to break up the aggregates or clusters of cells.
Disclosed is a disaggregating device for separating clusters of biological cells in a sample suspension, comprising: a beaker containing the suspension, a rotor rotatably mounted inside the beaker, a motor for rotating the rotor, the rotor being spaced apart from the walls of the beaker and rotated at a sufficient speed to create shear forces on the clusters of cells located in the sample suspension between the rotor and the beaker to break up the aggregates or clusters of cells.
Description
~78~;3 The present invention relates to disaggregation devices for cellular suspensions to prepare samples for subsequent analysis~
Disaggregation of clusters of cells in tissue samples, scrapes, and body fluids is often desirable for visual analysis and is of extreme importance for quantitative analysis, especially where automated devices are used. Different techniques, both mechanical and chemical, have been tried to disaggregate cell clusters. For certain applications, particularly in cell cultures, the chemical techniques work well. For many applications, nowever, these techniques are too aggressive. Mechanical method~ which have been used include shaking and stirring, forced filtration, homogenization, syringing~ and ultrasonic agitation, aa ~hown by the following articles: 1) Garcia, G.L., and Tolles, W.E., "Ul'~rasonic disaggregation of cell clusters", J. Histochem. Cytochem., 25:508, 1977; ~) Mayall, 8.H., "Monodisperse cell 9amples: the problem and posaible solutions", in The Automation of Uterine Cances Cytology, ed. by G.L. Wied, G.F. Bahr, and P.H.
Bartels, Tutorials of Cytology, Chicago 1976, p. 61; 3) Miller, F., "Cytopreparatory methods: collection snearing, staining, screening, reporting", Compendium of Cytopreparatory Techniques, ed. by C.M.
Keebler, J.W. Reagan, and G.L. Wied, Tutorials of Cytology, Chicago, 1976, p. 59; 4) Wheeless, L.L., Jr., and Onderdonk, M.A., "Preparation of clinical gynecologic specimens for automated analysis: an overview", J. Histochem. Cytochem., 22:522, 1974; and 5) Rosenthal, D.L., Stern, E., McLatchie, C., Wu, A., Lagasse, L.D., Wall, R., and Castleman, K.R., "A Simple Method of Producing a Monolayer of Cervical Cells for Digital Image Processing", Anal. Quant. Cytol., 1:8h, 1979.
,~
lL1~8~L~;3 Of all the possible methods described in these articles, syringing is without a doubt the most successful method of cell disaggregation discovered to date. With syringing the cell suspension is repeatedly forced through a syringe needle. The shear forces at the tip of the needle are strong enough to break apart clusters of cells.
The present invention is directed toward a disaggregation device for separating clusters of biological cells in a sample suspeQsion, the device comprising a beaker containing the sample suspension, a rotor rotatably mounted inside the beaker, and means for rotating said rotor at a sufficient speed to create shear forces on the clusters of cells located in the sample suspension between the rotor and the beaker. In one modification to the device, a concentric inner vial can be positioned at the botto~ of the beaker between the rotor and the inner wall of the beaker to permit collection of fractions of differing densities. In another modification of the device, an inner cavity can be Eormed in the rotor with a plurality of holes extending through the rotor 90 as to allow the centrifugal force to push liquid from the cavity through the holes. In yet another modiiication to the device, the rotor can be provided with a helical, worm-like outer surface and is positioned inside a cap having a plurality of large diameter holes positioned in an upper region and a plurality of s~all diameter holes positioned in a lower region.
By virtue of the device embodying the present invention, high yields of single cells are obtainable, with no substantial additional cellular damage due to the technique, in time periods much shorter than those previously obtainable by prior art techniques.
781~33 By way of example only, illustrative embodiments of the invention now will be described with reference to the accompanying drawings in which:
FIGURE 1 is a perspective view of a first embodiment of a disaggregation device embodying the present invention;
FIGURE 2 is a fragmented, perspective view of a first modification of the disaggregation device shown in FIGURE 1;
FIGURE 3 is a cross-sectional view of a second modification to the embodiment of FIGURE 1 wherein the cross-section is taken with 10~ resp2ct to sectional lines 3-3 in FIGURE 4;
FIGURE 4 is a cross-sectional, top view of the modification shown in FIGURR 3 taken along section lines 4-4; and FIGURR 5 is a side view of yet another modification of the disaggregation device of FIGURE 1.
A disaggregation device, generally shown by reference numeral 10, is illustrated in FIGURE 1 and is operable for disflggregflting or sepflrating clusters of cells in tissue ~amples, scrapes, and body fluids. The disaggregated cells cfln subsequently be used for numerous common purposes, such as layering the cells on microscope slides for subsequent visual analysis with microscopes or by analysis with automated pattern recognition systems and for flow-through systems.
The disaggregation device 10 comprises a drum-like rotor 12 that is operable for spinning inside a round beaker 14 or like b~c~k~r ; 25 container. The rotor 12 and break~F 14 preferably have cylindrical shapes, but can assume other shapes, such as cone configurations. The beaker i4 is adapted for rontaining the cell sample in a liquid suspension 16. The rotor 12 is rigidly coupled to an electric motor 7~ 83 18 by way of a sha~t 20. The electric motor in turn is electrically coupled to a power source (not shown) for energizing the electric motor to turn the rotor 12. The electric motor 18 has a housing 22 wi~h a neck portion 24 extending downward from the lower extremities of the housing. ~he neck portion is adapted to slidingly fit within the top of the beaker 14, so that the housing 22 will rest upon and be supported by the beaker 14. Preferably but not necessarily, one or more ]edges 23 are positioned on the top portion of the rotor 12 to assist in preventing the liquid suspension 16 from moving up the inner b~k~ ~
wall of the bIFa~e~ 14.
Preferably, but not necessarily, the beaker 1~ is formed of plastic, the rotor 12 i9 formed of plastic, the shaft 20 is formed of metal, and the housing 22 and neck portion 24 are plastic. As one illustrated set of dimensions, the surface of the rotor 12 is spaced apart from the interior wall of the beaker by approximately 4 millimeters. The diameter of the rotor 12 can be, for example, 17 millimeters and the interior diameter of the beaker can be 25 millimeters. The rotor 12 is spaced apart from the floor of the beaker 14 by 1 millimeter and the vertical height of the rotor 12 is 14.5 millimeters to the ledges 23, with the two ledges 23 adding another 5 millimeters oE height. The diameter of the ledges 23 are 20 millimeters. Typically, the liquid suspension 16 might be 9 milliliters, so that the liquid level extends above the rotor 12 by 3 millimeters. These values are only given as illustrative values, and such values can vary substantially. For instance, spacings in the range of one centimeter have been used between the rotor 12 and the beaker 14, resulting in the desired disaggregation of the cells. In operation, the rotor 12 spins, creating shear forces between the outer wall of the rotor 12 and the inner wall of the beaker 1~. These shear forces are sufficient to break apart cell clusters without 11 78~83 significantly damaging the cells themselves. It is believed that these shear forces are created in a region immediately adjacent the wall of the rotor 12. The rotor 12 can be rotated, for example, between 2000 to 6000 R.P.M.'s. Generally, the faster the rotation, the better the results, with rotational speeds below 2000 R.P.M.'s being generally unacceptable. Tests using the device show a significant decrease in the time required to disaggregate cells, as compared to the traditional "syringing" of a cell suspension. In application of the disaggregation device 10 to cervical samples, yields of 80 to 90 percent single cells were obtained, with no cellular damage. These results were obtained in 30 seconds of rotor use. Similar results by syringing required a disaggregation period of 10 to 15 minutes. The device 10 is wel]-suited for use within an automated sample preparation device. The rotor 12 is easily cleaned by operating it in a bath of running wat:er.
Although an elecCric motor 18 is shown in the pre~erred embodiment, other means of providing energy for rotating the rotor 12 are possible. For instance, a compact air turbine drive can be readily used in place of the small electric motor 18.
FIGURE 2 illustrates an adaptation of the disaggregation device 10 to permit collection of fractions of differing densities.
The beaker 14 is modified to form a double-compartmented beaker by the inclusion of a concentric inner vial 26. The inner vial 26 can extend to a position below or above the bottom of the rotor 12. PreEerably, the top of the inner vial 26 is positioned slightly below the bottom of the rotor 12. By selecting the intercompartmental wall placement and height of the vial 26, it is possible to collect components of the liquid suspension having differing densities, due to the centrifugal action of the rotor 12. Moreover, it is contemplated that with a plurality of concentric inner v`ials 26 provided along the floor of the ~L~l78 183 beaker 14, it will be possible to separate the sample into more than two fractions.
FIGURES 3 and 4 show another modification of the embodiment of FIGURE 1. The rotor 12 has a central annular cavity 28 formed therein with a plurality of holes 30 extending from the cavity 28 to an outer wall 32 of the rotor. Each of the holes 30 has an elongaeed cylindrical portion 34 and a truncated cone-shaped, flared portion 36.
Again, the wall 32 of the rotor 12 is spaced apart from the inner wall ~`
of the beaker 14 by a similar distance to that shown in FIGURE 1.
Each of the holes 30 is aligned so its center axis is tangent to an imaginary circle which is concentric with the shaft 20, as shown in FIGURE 4. The cells to be disaggregated are drawn from the cavity 28 through the holes 30 by the centrifugal force created by the rotating rotor 12. In other words, this arrangernent function9 as a centrifugal pump. In addition to providing the heretofore described shearing force between the outer wall 32 of the rotor 12 and ~he inner wall o the beaker 14, an additional shear force is provided as éhe cellular material jets through the holes 30. Hence, the movement of the cells through the holes 30 enhances the cells disaggregation caused by the shear force between walls.
FIGURE 5 shows an alternative embodiment of the disaggregation device 10 wherein the rotor 12 has a helical cutout 38 that defines a worm-like exterior configuration. The beaker 14 is provided with greater cross-sectional dimensions relative to the beaker sizes of the previous embodimenes. A closed sleeve or cap 40 is interposed between the rotor 12 and the beaker 14. More specifically, the electric motor 18 now rests upon the top of the cap 40. The cap 40 has a closed end 42 which sits in the bottom of the beaker 14. A plurality of small diameter holes 44 pass through the cap in its lower regions toward the lower end of the rotor 12. A
:IL178~3 plurality of large diameeer holes 46 pass through the cap 40 in a region in the vicinity of the top of the rotor 12. Preferably, bu~
not necessarily, a ledge 47 is mounted to the beaker 14 at a height in the vicinity of the top of the liquid sample to p~event the liquid suspension from moving up the walls. With the direction of the helical cutout 38 shown in FIGURE 5, the clockwise rotation, as seen from helow, of the rotor 12.forces liquid downward toward the bottom o~ the cap 40. by virtue of this arrangement, liquid from the beaker 14 is drawn through the large diameter holes 46, is forced downward toward the bottom of cap 40 and proceeds outward through the small diameter holes 44. In addition to the shear force created between worm-like shaped surface of the rotor 12 and the inner wall of the beaker 14, additional shear forces are created as the liquid squirts out of the small diameter holes 44. Morleover, compared to the arrangement of FIGURES 3 and 4, the rotation Oe the helical cutout 38 allows eor the development Oe substantially greater prcssures Eor forcing the cells through the holes 44. More specieically, the combination of the shear force of the rotor 12 and the holes 44 in the cap 40 provides excellent disaggregation with up to 10 percent improvement over the basic rotor device shown in FIGURE 1. This added pressure also can be useful for stripping the cytoplasm from cell nuclei.
Although particular embodiments of the invention have been shown and described herein, there is no intention to thereby limit the invention to the details of such embodiments. On the contrary, the intention is to cover all modifications, alternatives, embodiments, usages and equivalents of the subject invention as fall within the spirit and scope of the invention, specification and the appended claims.
.. : ... , . ~. -,
Disaggregation of clusters of cells in tissue samples, scrapes, and body fluids is often desirable for visual analysis and is of extreme importance for quantitative analysis, especially where automated devices are used. Different techniques, both mechanical and chemical, have been tried to disaggregate cell clusters. For certain applications, particularly in cell cultures, the chemical techniques work well. For many applications, nowever, these techniques are too aggressive. Mechanical method~ which have been used include shaking and stirring, forced filtration, homogenization, syringing~ and ultrasonic agitation, aa ~hown by the following articles: 1) Garcia, G.L., and Tolles, W.E., "Ul'~rasonic disaggregation of cell clusters", J. Histochem. Cytochem., 25:508, 1977; ~) Mayall, 8.H., "Monodisperse cell 9amples: the problem and posaible solutions", in The Automation of Uterine Cances Cytology, ed. by G.L. Wied, G.F. Bahr, and P.H.
Bartels, Tutorials of Cytology, Chicago 1976, p. 61; 3) Miller, F., "Cytopreparatory methods: collection snearing, staining, screening, reporting", Compendium of Cytopreparatory Techniques, ed. by C.M.
Keebler, J.W. Reagan, and G.L. Wied, Tutorials of Cytology, Chicago, 1976, p. 59; 4) Wheeless, L.L., Jr., and Onderdonk, M.A., "Preparation of clinical gynecologic specimens for automated analysis: an overview", J. Histochem. Cytochem., 22:522, 1974; and 5) Rosenthal, D.L., Stern, E., McLatchie, C., Wu, A., Lagasse, L.D., Wall, R., and Castleman, K.R., "A Simple Method of Producing a Monolayer of Cervical Cells for Digital Image Processing", Anal. Quant. Cytol., 1:8h, 1979.
,~
lL1~8~L~;3 Of all the possible methods described in these articles, syringing is without a doubt the most successful method of cell disaggregation discovered to date. With syringing the cell suspension is repeatedly forced through a syringe needle. The shear forces at the tip of the needle are strong enough to break apart clusters of cells.
The present invention is directed toward a disaggregation device for separating clusters of biological cells in a sample suspeQsion, the device comprising a beaker containing the sample suspension, a rotor rotatably mounted inside the beaker, and means for rotating said rotor at a sufficient speed to create shear forces on the clusters of cells located in the sample suspension between the rotor and the beaker. In one modification to the device, a concentric inner vial can be positioned at the botto~ of the beaker between the rotor and the inner wall of the beaker to permit collection of fractions of differing densities. In another modification of the device, an inner cavity can be Eormed in the rotor with a plurality of holes extending through the rotor 90 as to allow the centrifugal force to push liquid from the cavity through the holes. In yet another modiiication to the device, the rotor can be provided with a helical, worm-like outer surface and is positioned inside a cap having a plurality of large diameter holes positioned in an upper region and a plurality of s~all diameter holes positioned in a lower region.
By virtue of the device embodying the present invention, high yields of single cells are obtainable, with no substantial additional cellular damage due to the technique, in time periods much shorter than those previously obtainable by prior art techniques.
781~33 By way of example only, illustrative embodiments of the invention now will be described with reference to the accompanying drawings in which:
FIGURE 1 is a perspective view of a first embodiment of a disaggregation device embodying the present invention;
FIGURE 2 is a fragmented, perspective view of a first modification of the disaggregation device shown in FIGURE 1;
FIGURE 3 is a cross-sectional view of a second modification to the embodiment of FIGURE 1 wherein the cross-section is taken with 10~ resp2ct to sectional lines 3-3 in FIGURE 4;
FIGURE 4 is a cross-sectional, top view of the modification shown in FIGURR 3 taken along section lines 4-4; and FIGURR 5 is a side view of yet another modification of the disaggregation device of FIGURE 1.
A disaggregation device, generally shown by reference numeral 10, is illustrated in FIGURE 1 and is operable for disflggregflting or sepflrating clusters of cells in tissue ~amples, scrapes, and body fluids. The disaggregated cells cfln subsequently be used for numerous common purposes, such as layering the cells on microscope slides for subsequent visual analysis with microscopes or by analysis with automated pattern recognition systems and for flow-through systems.
The disaggregation device 10 comprises a drum-like rotor 12 that is operable for spinning inside a round beaker 14 or like b~c~k~r ; 25 container. The rotor 12 and break~F 14 preferably have cylindrical shapes, but can assume other shapes, such as cone configurations. The beaker i4 is adapted for rontaining the cell sample in a liquid suspension 16. The rotor 12 is rigidly coupled to an electric motor 7~ 83 18 by way of a sha~t 20. The electric motor in turn is electrically coupled to a power source (not shown) for energizing the electric motor to turn the rotor 12. The electric motor 18 has a housing 22 wi~h a neck portion 24 extending downward from the lower extremities of the housing. ~he neck portion is adapted to slidingly fit within the top of the beaker 14, so that the housing 22 will rest upon and be supported by the beaker 14. Preferably but not necessarily, one or more ]edges 23 are positioned on the top portion of the rotor 12 to assist in preventing the liquid suspension 16 from moving up the inner b~k~ ~
wall of the bIFa~e~ 14.
Preferably, but not necessarily, the beaker 1~ is formed of plastic, the rotor 12 i9 formed of plastic, the shaft 20 is formed of metal, and the housing 22 and neck portion 24 are plastic. As one illustrated set of dimensions, the surface of the rotor 12 is spaced apart from the interior wall of the beaker by approximately 4 millimeters. The diameter of the rotor 12 can be, for example, 17 millimeters and the interior diameter of the beaker can be 25 millimeters. The rotor 12 is spaced apart from the floor of the beaker 14 by 1 millimeter and the vertical height of the rotor 12 is 14.5 millimeters to the ledges 23, with the two ledges 23 adding another 5 millimeters oE height. The diameter of the ledges 23 are 20 millimeters. Typically, the liquid suspension 16 might be 9 milliliters, so that the liquid level extends above the rotor 12 by 3 millimeters. These values are only given as illustrative values, and such values can vary substantially. For instance, spacings in the range of one centimeter have been used between the rotor 12 and the beaker 14, resulting in the desired disaggregation of the cells. In operation, the rotor 12 spins, creating shear forces between the outer wall of the rotor 12 and the inner wall of the beaker 1~. These shear forces are sufficient to break apart cell clusters without 11 78~83 significantly damaging the cells themselves. It is believed that these shear forces are created in a region immediately adjacent the wall of the rotor 12. The rotor 12 can be rotated, for example, between 2000 to 6000 R.P.M.'s. Generally, the faster the rotation, the better the results, with rotational speeds below 2000 R.P.M.'s being generally unacceptable. Tests using the device show a significant decrease in the time required to disaggregate cells, as compared to the traditional "syringing" of a cell suspension. In application of the disaggregation device 10 to cervical samples, yields of 80 to 90 percent single cells were obtained, with no cellular damage. These results were obtained in 30 seconds of rotor use. Similar results by syringing required a disaggregation period of 10 to 15 minutes. The device 10 is wel]-suited for use within an automated sample preparation device. The rotor 12 is easily cleaned by operating it in a bath of running wat:er.
Although an elecCric motor 18 is shown in the pre~erred embodiment, other means of providing energy for rotating the rotor 12 are possible. For instance, a compact air turbine drive can be readily used in place of the small electric motor 18.
FIGURE 2 illustrates an adaptation of the disaggregation device 10 to permit collection of fractions of differing densities.
The beaker 14 is modified to form a double-compartmented beaker by the inclusion of a concentric inner vial 26. The inner vial 26 can extend to a position below or above the bottom of the rotor 12. PreEerably, the top of the inner vial 26 is positioned slightly below the bottom of the rotor 12. By selecting the intercompartmental wall placement and height of the vial 26, it is possible to collect components of the liquid suspension having differing densities, due to the centrifugal action of the rotor 12. Moreover, it is contemplated that with a plurality of concentric inner v`ials 26 provided along the floor of the ~L~l78 183 beaker 14, it will be possible to separate the sample into more than two fractions.
FIGURES 3 and 4 show another modification of the embodiment of FIGURE 1. The rotor 12 has a central annular cavity 28 formed therein with a plurality of holes 30 extending from the cavity 28 to an outer wall 32 of the rotor. Each of the holes 30 has an elongaeed cylindrical portion 34 and a truncated cone-shaped, flared portion 36.
Again, the wall 32 of the rotor 12 is spaced apart from the inner wall ~`
of the beaker 14 by a similar distance to that shown in FIGURE 1.
Each of the holes 30 is aligned so its center axis is tangent to an imaginary circle which is concentric with the shaft 20, as shown in FIGURE 4. The cells to be disaggregated are drawn from the cavity 28 through the holes 30 by the centrifugal force created by the rotating rotor 12. In other words, this arrangernent function9 as a centrifugal pump. In addition to providing the heretofore described shearing force between the outer wall 32 of the rotor 12 and ~he inner wall o the beaker 14, an additional shear force is provided as éhe cellular material jets through the holes 30. Hence, the movement of the cells through the holes 30 enhances the cells disaggregation caused by the shear force between walls.
FIGURE 5 shows an alternative embodiment of the disaggregation device 10 wherein the rotor 12 has a helical cutout 38 that defines a worm-like exterior configuration. The beaker 14 is provided with greater cross-sectional dimensions relative to the beaker sizes of the previous embodimenes. A closed sleeve or cap 40 is interposed between the rotor 12 and the beaker 14. More specifically, the electric motor 18 now rests upon the top of the cap 40. The cap 40 has a closed end 42 which sits in the bottom of the beaker 14. A plurality of small diameter holes 44 pass through the cap in its lower regions toward the lower end of the rotor 12. A
:IL178~3 plurality of large diameeer holes 46 pass through the cap 40 in a region in the vicinity of the top of the rotor 12. Preferably, bu~
not necessarily, a ledge 47 is mounted to the beaker 14 at a height in the vicinity of the top of the liquid sample to p~event the liquid suspension from moving up the walls. With the direction of the helical cutout 38 shown in FIGURE 5, the clockwise rotation, as seen from helow, of the rotor 12.forces liquid downward toward the bottom o~ the cap 40. by virtue of this arrangement, liquid from the beaker 14 is drawn through the large diameter holes 46, is forced downward toward the bottom of cap 40 and proceeds outward through the small diameter holes 44. In addition to the shear force created between worm-like shaped surface of the rotor 12 and the inner wall of the beaker 14, additional shear forces are created as the liquid squirts out of the small diameter holes 44. Morleover, compared to the arrangement of FIGURES 3 and 4, the rotation Oe the helical cutout 38 allows eor the development Oe substantially greater prcssures Eor forcing the cells through the holes 44. More specieically, the combination of the shear force of the rotor 12 and the holes 44 in the cap 40 provides excellent disaggregation with up to 10 percent improvement over the basic rotor device shown in FIGURE 1. This added pressure also can be useful for stripping the cytoplasm from cell nuclei.
Although particular embodiments of the invention have been shown and described herein, there is no intention to thereby limit the invention to the details of such embodiments. On the contrary, the intention is to cover all modifications, alternatives, embodiments, usages and equivalents of the subject invention as fall within the spirit and scope of the invention, specification and the appended claims.
.. : ... , . ~. -,
Claims (8)
1. A disaggregation device for separating clusters of biological cells in a sample suspension, comprising: a beaker containing said sample suspension; a rotor rotatably mounted inside said beaker; and rotating means for rotating said rotor to create shear forces to disaggregate said clusters of cells located in the sample suspension between said rotor and said beaker.
2. The disaggregation device according to claim 1, wherein the outer surface of said rotor and the inner wall of said beaker have annular cross-sectional configurations.
3. The disaggregation device according to claim 1 wherein said rotor has a lower end which is positioned in spaced-apart relationship to the bottom of said beaker.
4. The disaggregation device according to any one of claims 1, 2 or 3, further including an inner vial positioned between said rotor and said beaker at the bottom of said beaker.
5. The disaggregation device according to any one of claims 1, 2 or 3, further including an inner vial positioned between said rotor and said beaker at the bottom of said beaker, wherein said inner vial has a circular cross-sectional configuration and is positioned in concentric relationship with said beaker and said rotor, and said inner vial has a height that extends upward proximate to the vicinity of the lower end of said rotor.
6. The disaggregation device according to any one of claims 1, 2 or 3, in which: said rotor has a cavity formed therein; a plurality of holes pass through said rotor, and said holes extend from said cavity to the outer surface of said rotor; whereby the rotation of said rotor creates a centrifugal force that moves the sample from said cavity through said holes.
7. The disaggregation device according to any one of claims 1, 2 or 3, further comprising: a cap positioned inside of said beaker; said cap having a plurality of relatively small diameter holes and a plurality of relatively large diameter holes, all passing through the wall of said cap, with said large diameter holes being positioned above said small diameter holes; and said rotor having on its surface a helical cutout; whereby rotation of said rotor pulls the sample suspension through said large diameter holes and pushes the sample suspension downward inside of said cap so as to force the sample suspension through said small diameter holes.
8. The disaggregation device according to any one of claims 1, 2 or 3, further comprising: a cap positioned inside of said beaker; said cap having a plurality of relatively small diameter holes and a plurality of relatively large diameter holes, all passing through the wall of said cap, with said large diameter holes being positioned above said small diameter holes; said large diameter holes being located at or above the upper portion of said rotor and said small diameter holes being located at or below the lower portion of said rotor; and said rotor having on its surface a helical cutout;
whereby rotation of said rotor pulls the sample suspension through said large diameter holes and pushes the sample suspension downward inside of said cap so as to force the sample suspension through said small diameter holes.
whereby rotation of said rotor pulls the sample suspension through said large diameter holes and pushes the sample suspension downward inside of said cap so as to force the sample suspension through said small diameter holes.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US26371181A | 1981-05-14 | 1981-05-14 | |
| US06/263,711 | 1981-05-14 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1178183A true CA1178183A (en) | 1984-11-20 |
Family
ID=23002939
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000402150A Expired CA1178183A (en) | 1981-05-14 | 1982-05-03 | Disaggregation device for cell suspensions |
Country Status (6)
| Country | Link |
|---|---|
| JP (1) | JPS57194355A (en) |
| CA (1) | CA1178183A (en) |
| DE (1) | DE3218079A1 (en) |
| FR (1) | FR2505870A1 (en) |
| GB (1) | GB2100137B (en) |
| NL (1) | NL8201972A (en) |
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| US5143627A (en) * | 1990-07-09 | 1992-09-01 | Cytyc Corporation | Method and apparatus for preparing cells for examination |
| US5168066A (en) * | 1990-03-07 | 1992-12-01 | Cytyc Corporation | Thionin staining and imaging technique |
| US5240606A (en) * | 1990-07-09 | 1993-08-31 | Cytyc Corporation | Apparatus for preparing cells for examination |
| US5269918A (en) * | 1990-07-09 | 1993-12-14 | Cytyc Corporation | Clinical cartridge apparatus |
| US5282978A (en) * | 1990-07-09 | 1994-02-01 | Cytyc Corporation | Specimen processor method and apparatus |
| US6010909A (en) * | 1990-03-02 | 2000-01-04 | Cytyc Corporation | Method and apparatus for controlled instrumentation of particles with a filter device |
| US6318190B1 (en) | 1996-11-01 | 2001-11-20 | Cytyc Corporation | Systems for collecting fluid samples having select concentrations of particles |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4937187A (en) * | 1983-02-04 | 1990-06-26 | Brown University Research Foundation | Methods for separating malignant cells from clinical specimens |
| DE3427114A1 (en) * | 1984-07-23 | 1986-01-30 | A. Dr.med. 8022 Grünwald Szabados | Sample receptacle for paste-like sample material and method for processing paste-like sample material |
| US4678559A (en) * | 1984-07-23 | 1987-07-07 | Andreas Szabados | Test specimen container for pasty specimen material |
| US4828395A (en) * | 1985-02-21 | 1989-05-09 | Yamato Scientific Company, Limited | Continuous flow type homogenizer |
| DE19729028C1 (en) | 1997-07-08 | 1999-05-06 | Fraunhofer Ges Forschung | Device and method for isolating cell material from a tissue association and / or a liquid |
| IE20030856A1 (en) * | 2003-11-14 | 2005-06-15 | Enfer Technology Ltd | Sample homogeniser |
| DE102007005369A1 (en) * | 2007-02-02 | 2008-08-07 | Eppendorf Ag | Device and method for the mechanical separation of cells from a cell network |
-
1982
- 1982-05-03 CA CA000402150A patent/CA1178183A/en not_active Expired
- 1982-05-13 GB GB8213982A patent/GB2100137B/en not_active Expired
- 1982-05-13 FR FR8208356A patent/FR2505870A1/en active Granted
- 1982-05-13 JP JP7926882A patent/JPS57194355A/en active Pending
- 1982-05-13 DE DE19823218079 patent/DE3218079A1/en not_active Withdrawn
- 1982-05-13 NL NL8201972A patent/NL8201972A/en not_active Application Discontinuation
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6010909A (en) * | 1990-03-02 | 2000-01-04 | Cytyc Corporation | Method and apparatus for controlled instrumentation of particles with a filter device |
| US5168066A (en) * | 1990-03-07 | 1992-12-01 | Cytyc Corporation | Thionin staining and imaging technique |
| US5143627A (en) * | 1990-07-09 | 1992-09-01 | Cytyc Corporation | Method and apparatus for preparing cells for examination |
| US5240606A (en) * | 1990-07-09 | 1993-08-31 | Cytyc Corporation | Apparatus for preparing cells for examination |
| US5269918A (en) * | 1990-07-09 | 1993-12-14 | Cytyc Corporation | Clinical cartridge apparatus |
| US5282978A (en) * | 1990-07-09 | 1994-02-01 | Cytyc Corporation | Specimen processor method and apparatus |
| US6318190B1 (en) | 1996-11-01 | 2001-11-20 | Cytyc Corporation | Systems for collecting fluid samples having select concentrations of particles |
| US6634244B2 (en) | 1996-11-01 | 2003-10-21 | Cytyc Corporation | Methods for collecting fluid samples having select concentrations of particles |
Also Published As
| Publication number | Publication date |
|---|---|
| FR2505870A1 (en) | 1982-11-19 |
| NL8201972A (en) | 1982-12-01 |
| GB2100137A (en) | 1982-12-22 |
| FR2505870B3 (en) | 1984-04-06 |
| DE3218079A1 (en) | 1982-12-02 |
| GB2100137B (en) | 1985-10-23 |
| JPS57194355A (en) | 1982-11-29 |
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