Method and apparatus for centrifugation-based accumulation and collection of cell cultures

Abstract

A cover lid collector (10) for a sample carrier (40) for cell cultivation, wherein the cover lid collector (10) comprises a connecting portion (12) for engaging with the sample carrier (40) for closing the same, wherein the connecting portion (12) defines a first plane (11). The cover lid collector (10) further comprises a cavity (14), which when the cover lid collector is engaged with the sample carrier (40), has a cross-section that decreases in a direction perpendicular to the first plane (11) and pointing away from said sample carrier (40) between a near end (16) of the cavity (14) and a far end (18) of the cavity (14), wherein the near end (16) is closer to the sample carrier (40) than the far end (18).

Claims

1. A cover lid collector for a sample carrier for cell cultivation, wherein the cover lid collector comprises: a connecting portion for engaging with the sample carrier for closing the same, wherein the connecting portion defines a first plane; and a cavity, which when the cover lid collector is engaged with the sample carrier, has a cross-section that decreases in a direction perpendicular to the first plane and pointing away from said sample carrier between a near end of the cavity and a far end of the cavity, wherein the near end is closer to the sample carrier than the far end.

2. The cover lid collector of claim 1, wherein the cross-section of the cavity decreases monotonically in said direction perpendicular to the first plane and pointing away from said sample carrier between a near end of the cavity and a far end of the cavity.

3. The cover lid collector of claim 2, wherein the cavity is formed by a plurality of planar walls (20a-20d) that extend between the near end and the far end, such that the cross-section of the cavity has a polygonal shape.

4. The cover lid collector of claim 3, wherein at least one of the planar walls has a polygonal form, preferably a triangular form.

5. The cover lid collector of claim 3, wherein one or more of the planar walls has a slope angle that is different to the slope angle of another one of the planar walls, wherein the slope angle is an angle defined between the surface of the wall and the first plane.

6. The cover lid collector of claim 2, wherein the planar walls have a common vertex that forms the far end of the cavity.

7. The cover lid collector of claim 6, wherein the cavity is formed by four triangular shaped walls, wherein said four walls are pairwise adjacent and opposed, such that the cavity has a symmetric or asymmetric quasipyramidal shape.

8. The cover lid collector of claim 1, further comprising a drain opening that fluidly connects the far end of the cavity to the exterior of the cover lid collector.

9. The cover lid collector of claim 8, further comprising a sealing cap configured for tightly closing the drain opening.

10. The cover lid collector of claim 1, further comprising a sealing element configured for being arranged around the cover lid collector and around the sample carrier so as to provide a tight sealed connection between the cover lid collector and the sample carrier when the cover lid collector is engaged with the sample carrier.

11. The cover lid collector of claim 1, wherein the cavity or at least one of the planar walls thereof are partly or fully coated with a magnetic material, with an antiadhesive coating, or with an adhesive coating.

12. (canceled)

13. (canceled)

14. The cover lid collector of claim 1, further comprising a removable separating foil covering the near end of the cavity.

15. The cover lid collector of claim 1, wherein the cover lid collector is totally or partially made of a transparent material, such that the interior of the cavity can be seen from the outside of the cover lid collector.

16. The cover lid collector of claim 1, wherein the cover lid collector comprises one or more cavities.

17. A method of separating and extracting biological samples comprised in a liquid medium, the liquid medium being contained in a sample carrier for cell cultivation, using a cover lid collector that comprises: a connecting portion for engaging with the sample carrier for closing the same, wherein the connecting portion defines a first plane; and a cavity, which when the cover lid collector is engaged with the sample carrier, is in fluid communication with at least some of the biological samples, and has a cross-section that decreases in a direction perpendicular to the first plane and pointing away from said sample carrier between a near end of the cavity and a far end of the cavity, wherein the near end is closer to the sample carrier than the far end, wherein the method comprises the steps of: engaging the cover lid collector with the sample carrier, such that the cover lid collector is tightly connected to the sample carrier; placing the cover lid collector engaged with the sample carrier in a centrifuge, such that the cover lid collector is positioned over the sample carrier; centrifuging the cover lid collector engaged with the sample carrier using the centrifuge such as to subject the liquid medium and the biological samples to a centrifugal force in said direction, such that the biological samples form a first phase and the liquid medium forms a second phase, wherein the first phase and the second phase are separated from each other and wherein the first phase is closer to the far end of the cavity than the second phase; and extracting the biological sample from the cover lid collector.

18. The method of claim 17, wherein the cover lid collector further comprises a drain opening that fluidly connects the far end of the cavity to the exterior of the cover lid collector, wherein extracting comprises extracting the second phase through the drain opening.

19. The method of claim 17, wherein centrifuging comprises centrifuging at different centrifugation speeds.

20. The method of claim 17, wherein the sample carrier comprises one of a multi-well plate and a hanging-drop plate.

21. The method of claim 20, further comprising, after engaging the cover lid collector with the multi-well plate and before placing the cover lid collector engaged with the multi-well plate in the centrifuge, a step of turning the cover lid collector engaged with the multi-well plate, such that the multi-well plate is located above the cover lid collector.

22. (canceled)

23. A cover lid collector for a sample carrier for cell cultivation, wherein the cover lid collector comprises: a connecting portion for engaging with the sample carrier for closing the same, wherein the connecting portion defines a first plane; and a cavity, which when the cover lid collector is engaged with the sample carrier, has a cross-section that decreases in a direction perpendicular to the first plane and pointing away from said sample carrier between a near end of the cavity and a far end of the cavity, wherein the near end is closer to the sample carrier than the far end; wherein the cavity is formed by a plurality of planar walls that extend between the near end and the far end, such that the cross-section of the cavity has a polygonal shape.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0050] FIG. 1 shows different views of a cover lid collector according to an embodiment of the invention.

[0051] FIG. 2 shows different views of a cover lid collector according to a different embodiment of the invention.

[0052] FIG. 3 shows different views of the engagement of a cover lid collector according to an embodiment of the invention comprising a sealing element with a sample carrier.

[0053] FIG. 4 shows different views of the a cover lid collector according to an embodiment of the invention comprising a removable separating foil.

[0054] FIG. 5 shows different views of a cover lid collector according to another embodiment of the invention.

[0055] FIG. 6 shows a flow diagram illustrating a method according to an embodiment of the invention.

[0056] FIG. 7 shows different schematic views of the use of a cover lid collector according to the method illustrated in FIG. 6.

[0057] FIG. 8 shows a flow diagram illustrating a method according to a different embodiment of the invention.

[0058] FIG. 9 shows different schematic views of the use of a cover lid collector according to the method illustrated in of FIG. 8.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0059] For the purposes of promoting an understanding of the principles of the invention, reference will now be made to a preferred embodiment illustrated in the drawings, and specific language will be used to describe the same. It will nevertheless be understood that no limitation of the scope of the invention is thereby intended, such alterations and further modifications in the illustrated apparatus and such further applications of the principles of the invention as illustrated therein being contemplated as would normally occur now or in the future to one skilled in the art to which the invention relates.

[0060] FIG. 1 shows a top view (A), a perspective view (B) and a side view (C) of a cover lid collector to according to an embodiment of the invention. The cover lid collector 10 is configured for engaging with a sample carrier for cell cultivation, which is not shown in the figure. The cover lid collector 10 comprises a connecting portion 12 for engaging with the sample carrier so as to close the sample carrier with the cover lid collector to. For this purpose, the size, dimensions and shape of the cover lid collector 10, and in particular of the connecting portion 12 thereof match those of the sample carrier with which the cover lid collector 10 is to be engaged. In the embodiment shown, the cover lid collector 10 is tailored to a sample carrier that corresponds to a Falcon 96 Well Clear Flat Bottom TC-Treated Culture Microplate and has a maximum length of 127.63 mm and a maximal width of 85.11 mm, corresponding to the dimensions necessary for being tightly connected to the aforesaid sample carrier. The connecting portion 12 defines a first plane 11.

[0061] The cover lid collector 10 further comprises a cavity 14. When the cover lid collector 10 is engaged with the sample carrier, the cavity 14 is in communication with those parts of the sample carrier that are configured for receiving, supporting or containing samples. The cross-section of the cavity 14, defined as the area of a plane parallel to the first plane 11 intersected by the cavity 14, decreases in a direction perpendicular to the first plane 11 that runs between a near end 16 of the cavity 14 and a far end 18 of the cavity 14 away from the sample carrier, that is, away from the first plane 11. This direction is indicated in FIG. 1C by the vertical arrow 5. In the embodiment shown, the cross-section of the cavity 14 decreases monotonically in the direction indicated by the arrow 5.

[0062] In the embodiment shown, the cavity 14 is formed by four planar walls 20a, 20b, 20c, and 20d, which extend between the near end 16 and the far end 18 of the cavity 14. The cross-section of the cavity 14 hence has a polygonal rectangular shape. In the embodiment shown, the cover lid collector 10 comprises an optional indented corner that allows identifying a particular orientation thereof. The planar walls 20c and 20d have a triangular shape, whereas the planar walls 20a and 20b have an irregular quadrilateral shape. The planar walls 20a 20c are opposed to each other, and so are the planar walls 20b and 20d. Further, the planar wall 20a is adjacent to the planar wall 20b and to the planar wall 20d, and so is the planar wall 20c.

[0063] The angle of each of the planar walls 20a-20d with the first plane 11 is referred to as the slope angle there. In the embodiment shown, the planar walls 20a and 20c both have a first slope angle of 20, the planar wall 20b has a second slope angle of 7, and the planar wall 20d has a third slope angle of 56. Thus, the cavity 14 displays an asymmetric quasipyramidal shape. The planar walls 20a-20d have a common vertex that forms the far end 18 of the cavity 14. In the embodiment shown, the cover lid collector 10 is made of polystyrene and the planar walls 20a-20d are coated with an antiadhesive coating.

[0064] FIG. 2 shows a top view (A), a perspective view (B) and a side view (C) of a cover lid collector 10 according to another embodiment of the invention that comprises all elements described with respect to the embodiment shown in FIG. 1, which are not described here again. In the embodiment shown in FIG. 2, the cover lid collector 10 further comprises a drain opening 30 arranged at the far end 18 of the cavity 14, which far end 18 is formed by the common vertex of the planar walls 20a-20d. The drain opening 30 fluidly connects the interior of the cavity 14 to the exterior of the cover lid collector 10 through the planar surface 13 of the cover lid collector 10, which is parallel to the first plane 11 and separated from it by the cavity 14. The cover lid collector 10 further comprises a sealing cap 32, which is configured for tightly closing the dry opening 30.

[0065] FIG. 3 shows different views of the engagement of a cover lid collector 10 according to an embodiment of the invention with a multi-well plate acting as a sample carrier 40, wherein the cover lid collector 10 further comprises a sealing element 42 that is shown separately in FIG. 3A. The sealing element 42 is configured for being arranged around the cover lid collector 10 and around the sample carrier 40 as illustrated in the exploded view shown in FIG. 3B. The sealing element 42 provides a tight sealed connection between the cover lid collector 10 and the sample carrier 40 when the cover lid collector 10 is engaged with the sample carrier 40 as shown in FIG. 3C.

[0066] The sealing element 42 shown in the embodiment of FIG. 3 has a band-like-shape and is made of rubber. The sealing element 42 has a perimeter that exceeds that of the connecting portion 12 of the cover lid collector 10 by 3 mm, whereby a tight connection between the cover lid collector 10 and the sample carrier 10 is possible even when the dimensions of the connecting portion 12 of the cover lid collector 10 do not exactly match those of the sample carrier 40.

[0067] FIG. 4 shows a top view of a cover lid collector 10 according to an embodiment of the invention that comprises a removable separating foil 44 that is configured for covering the near end 16 of the cavity 14 of the cover lid collector 10. The removable separating foil 44 prevents leakage of the samples contained in a sample carrier 40 into the cavity 14 of the cover lid collector 10 when manipulating the cover lid collector 10 and the sample carrier 40 before centrifugation, i.e. in the situation shown in FIG. 4A. The strength of the removable separating foil 44 is chosen such that it may be disrupted or broken by centrifugal forces upon centrifugation, such that once centrifugal forces set in, as shown in FIG. 4B, the removable separating foil 44 no longer blocks access to the cavity 14 for the samples contained in a sample carrier 40 that is engaged with the cover lid collector 10 such that the samples may enter the cavity 14 during centrifugation.

[0068] FIGS. 5A and 5B respectively show a top view and a side view of a cover lid collector 10 according to an embodiment of the invention that comprises four cavities 14a, 14b, 14c and 14d. When the cover lid collector 10 is engaged with a sample carrier 40, which in the example shown in FIG. 4B corresponds to a multi-well plate, first to fourth parts of the sample carrier 40, which in the embodiment shown respectively correspond to first to fourth groups of wells of the multi-well plate and may respectively contain first to fourth parts of the samples contained in the sample carrier 40, are respectively in communication with the first to fourth cavities 14a to 14d. In the embodiment shown, the first part of the samples are contained in the wells comprised in said first part of the multi-well plate 40, the second part of the samples are contained in the wells comprised in the second part of the multi-well plate 40, and so forth.

[0069] When the sample carrier 40 and the cover lid collector 10 are engaged with each other and subjected to centrifugation in order to separate the biological samples from the liquid medium in each of the samples contained in the sample carrier 40, the biological samples comprised in the samples contained in the first part of the sample carrier 40 are collected at the far end of the first cavity 14a, the biological samples comprised in the samples contained in the second part of the sample carrier 40 are collected at the far end of the second cavity 14b, and so forth. In the embodiment shown, the first to fourth cavities 14a to 14d all have a regular pyramidal shape, but other embodiments may comprise cavities of different forms and sizes. In particular, the form and shape of the different cavities may be adapted to the form and shape of a sample carrier or to a distribution pattern of the samples in a sample carrier.

[0070] FIG. 6 shows a flow diagram of a method 100 according to an embodiment of the invention of separating and extracting biological samples comprised in a liquid medium contained in a sample carrier 40 corresponding to a multi-well plate 40, wherein the biological samples and to the liquid medium constitute samples in the form of a fluid sample that is contained in the wells of the multi-well plate 40. The method 100 foresees the use of a cover lid collector to according to any of the embodiments of the present invention described above.

[0071] The method 100 comprises a step 102 of engaging the cover lid collector 10 with the multi-well plate 40. As a result of method step 102, the cover lid collector 10 is tightly connected to the multi-well plate 40 by means of the connecting portion 12.

[0072] In method step 104, the cover lid collector 10 engaged with the multi-well plate 40 is turned upside down, such that the multi-well plate 40 is now located over the cover lid collector 10. The cover lid collector 10 engaged with the multi-well plate 40 are placed in the centrifuge 50 in this orientation, which is opposite to the usual orientation of a sample carrier in a centrifuge.

[0073] The method too further comprises a step 106 of placing the cover lid collector 10 engaged with the multi-well plate 40 in a centrifuge 50 such that it may be centrifuged by means of the centrifuge 50

[0074] In method step 108, the cover lid collector 10 engaged with the sample carrier 40 is centrifuged using the centrifuge 50. As a consequence of centrifugation, biological samples comprised in the samples contained in the sample carrier 40, i.e. contained in the wells of the multi-well plate 40, are separated from the liquid medium in which they were previously distributed. As explained in detail above, as a consequence of the centrifugation with the cover lid collector 10, the biological samples form a first phase and the liquid medium forms a second phase, wherein the first phase and the second phase are separated from each other, and wherein the first phase is closer to the far end 18 of the cavity 14 of the cover lid collector 10 than the second phase.

[0075] In method step 110, the cover lid collector 10 engaged with the sample carrier is removed from the centrifuge 50 after having been subject to centrifugation therein.

[0076] The method too then continues with step 112, in which biological samples present in the first phase, which is concentrated at the far end 18 of the cavity 14, are extracted from the cover lid collector 10 e.g. by aspirating the biological samples with a laboratory aspirator or by pipetting.

[0077] FIGS. 7A to 7E show different schematic views of a cover lid collector 10 according to an embodiment of the invention, a sample carrier 40 corresponding to a multi-well plate 40, and a centrifuge 50, which are used according to the method 100 of FIG. 6 of separating and to extracting biological samples comprised in a liquid medium, wherein the biological samples and the liquid medium constitute samples in the form of a fluid sample that is contained in the wells of the multi-well plate 40.

[0078] FIG. 7A corresponds to snapshot representative of method step 102 in which the cover lid collector 10 is engaged with the multi-well plate 40.

[0079] FIG. 7B corresponds to snapshot representative of method step 104 of turning the cover lid collector 10 engaged with the multi-well plate 40 upside down such that they have the orientation shown in the figure in which the multi-well plate 40 is positioned above the cover lid collector 10.

[0080] FIG. 7C corresponds to snapshot representative of method step 106 of placing the cover lid collector 10 engaged with the multi-well plate 40 in a centrifuge 50. In the case of the centrifuge 50 shown in FIG. 7, which is an Eppendorf 5810R Centrifuge with Rotor Packages supplemented with a swing-bucket rotor with four MTP/Flex carriers, method step 106 comprises placing the cover lid collector 10 engaged with the multi-well plate 40 on a swing-out plate carrier 52 of the centrifuge 50.

[0081] FIG. 7D corresponds to snapshot representative of method step 108 of centrifuging the cover lid collector 10 engaged with the multi-well plate 40 in the centrifuge 50 such that centrifugal forces arise in the direction normal to the cover lid collector 10 pointing away from the multi-well plate 40, i.e. pointing towards the far end 18 of the cavity 14, such that biological samples accumulate at the far end 18 of the cavity 14 of the cover lid collector 10.

[0082] After centrifugation, the cover lid collector 10 and the multi-well plate 40 are removed from the centrifuge 50 and method step 112 of extracting the biological samples from the cover lid collector 10 is carried out. For this purpose, in the embodiment shown, the cover lid collector 10 and the multi-well plate 40 are disengaged while keeping the orientation shown in FIG. 7B, as shown in FIG. 7E.

[0083] FIG. 8 shows a flow diagram of a method 200 according to another embodiment of the invention of separating and extracting biological samples comprised in a liquid medium contained in a sample carrier 40 corresponding to a hanging drop plate 40, wherein the biological samples and the liquid medium constitute samples in the form of drops that are contained in the hanging drop plate 40. The method 200 foresees the use of a cover lid collector 10 according to any of the embodiments of the present invention described above.

[0084] The method 200 comprises a step 202 of engaging the cover lid collector 10 with the hanging drop plate 40. After method step 202, the cover lid collector 10 is tightly connected to the hanging drop plate 40 by means of the connecting portion 12 of the cover lid collector 10.

[0085] In method step 204, the cover lid collector 10 engaged with the hanging drop plate 40 is placed in a centrifuge 50 such that it may be centrifuged by means of the centrifuge 50. In the embodiment shown, this comprises placing the cover lid collector 10 engaged with the hanging drop plate 40 on a swing-out plate carrier 52 the centrifuge 50, which also corresponds to an Eppendorf 5810R Centrifuge with Rotor Packages supplemented with a swing-bucket rotor with four MTP/Flex carriers.

[0086] In method step 206, the cover lid collector 10 engaged with the hanging drop plate 40 is centrifuged using the centrifuge 50. As a result of centrifugation, biological samples comprised in the samples, i.e. in the hanging drops, contained in the hanging drop plate are separated from the liquid medium in which they were previously distributed in a way analogous to method step 108 with reference to FIGS. 6 and 7. The biological samples then form a first phase and the liquid medium forms a second phase, wherein the first phase and the second phase are separated from each other, and wherein the first phase is closer to the far end 18 of the cavity 14 of the cover lid collector 10 than the second phase.

[0087] In method step 208, the cover lid collector 10 engaged with the hanging drop plate is removed from the centrifuge 50.

[0088] In method step 210, the biological samples present in the first phase, which is concentrated at the far end 18 of the cavity 14 of the cover lid collector 10, are extracted from the cover lid collector 10, e.g. by aspirating the biological samples with a laboratory aspirator or by pipetting.

[0089] FIGS. 9A to 9D show different schematic views of a cover lid collector 10 according to another embodiment of the invention, a sample carrier 40 corresponding to a hanging drop plate 40, and a centrifuge 50, which are used according to the method 200 of FIG. 8 of separating and extracting biological samples comprised in a liquid medium, wherein the biological samples and the liquid medium constitute samples in the form of drops contained in the hanging drop plate 40.

[0090] FIG. 9A corresponds to a snapshot representative of method step 202 in which the cover lid collector 10 is engaged with the hanging drop plate 40, wherein the cover lid collector is below the hanging drop plate 40.

[0091] FIG. 9B corresponds to snapshot representative of method step 204 of placing the cover lid collector 10 engaged with the hanging drop plate 40 on one of the swing-out plate carriers 52 of a centrifuge 50 in the orientation shown in FIG. 9A.

[0092] FIG. 9C corresponds to snapshot representative of method step 206 of centrifuging the cover lid collector 10 engaged with the hanging drop plate 40 in the centrifuge 50 such that centrifugal forces arise in the direction normal to the cover lid collector 10 pointing away from the hanging drop plate 40, i.e. pointing towards the far end 18 of the cavity 14, such that biological samples accumulate at the far end 18 of the cavity 14 of the cover lid collector 10.

[0093] After centrifugation, the cover lid collector 10 and the multi-well plate 40 are removed from the centrifuge 50 and method step 210 of extracting the biological samples from the cover lid collector 10 is carried out. For this purpose, in the embodiment shown, the cover lid collector 10 and the hanging drop plate 40 can be disengaged while keeping the orientation shown in FIG. 9B, as shown in FIG. 9D.

[0094] Although preferred exemplary embodiments are shown and specified in detail in the drawings and the preceding specification, these should be viewed as purely exemplary and not as limiting the invention. It is noted in this regard that only the preferred exemplary embodiments are shown and specified, and all variations and modifications should be protected that presently or in the future lie within the scope of protection of the invention as defined in the claims.

REFERENCE SIGN LIST

[0095] 10 cover lid collector [0096] 11 first plane defined by connecting portion [0097] 12 connecting portion [0098] 14 cavity [0099] 14a-14d cavities [0100] 16 near end of the cavity [0101] 18 far end of the cavity [0102] 20a-20d planar walls of the cavity [0103] 30 drain opening [0104] 32 sealing cap [0105] 40 sample carrier [0106] 42 sealing element [0107] 44 removable separating foil [0108] 50 centrifuge [0109] 52 swing-out plate carrier