Abstract
The present invention relates to a means for improving fluid exchange across a microplate, comprising a microplate which comprises one or more internal modifications which aid the introduction and removal of fluids from the wells of the plate and minimize the damage caused to biological matter located therein upon introduction or removal of a pipette tip and/or fluids, and/or a means for inclining the microplate at an angle from the horizontal and retaining the microplate in the inclined position, to facilitate fluid introduction and aspiration from the wells of the microplate.
Claims
1-35. (canceled)
36. A microplate comprising a plurality of open wells, wherein one or more of the wells comprises an area for communication with the tip of a pipette which provides a point of contact between the dispensing end of the pipette tip and the microplate well in use, wherein the area for communication with the pipette tip is located at the junction between the base of the well and the wall of the well and wherein the area for communication with the pipette tip comprises one or more protrusions located on the base and/or wall of the well interior, and/or one or more recesses in the wall and/or base of the well.
37. The microplate of claim 36, wherein the area for communication with the pipette tip is such that, in use, the dispensing end of the pipette tip bridges the gap between a point on the base of the microplate well and a point on the wall of the microplate well to create a channel for aspiration and dispensing of fluid.
38. The microplate of claim 36, wherein an area for communication with the pipette tip comprises two or more raised areas which are located at a distance from each other which is comparable to a diameter of an appropriately sized pipette tip for an assay using the microplate.
39. The microplate of claim 36, wherein the protrusions and the recesses prevent lateral movement of the pipette tip.
40. The microplate of claim 36, wherein there are two areas for communication with the pipette tip, which are located at opposite sides of the microplate well.
41. The microplate of claim 36, wherein the one or more wells further comprises at least one guide channel.
42. The microplate of claim 36, wherein the end of the guide channel or guide channels are in close proximity to or communicate with the area for communication with the pipette tip.
43. The microplate of claim 41, wherein the guide channel accommodates a pipette tip of a size appropriate for an assay using the microplate.
44. The microplate of claim 41, wherein the guide channel comprises an indentation in the interior wall of the well, or one or more raised areas on the interior wall of the well, and/or terminates in an indentation in the wall of the microplate well.
45. A system comprising a combination of a microplate according to claim 36 and a microplate holder, wherein the microplate holder comprises a means for inclining a microplate, so that the microplate is positioned so that one aspect, side or edge of the microplate is vertically raised relative to another aspect, side or edge of the microplate.
46. The system of claim 45, wherein the microplate is at an angle from a conventional horizontal orientation, an angle of inclination of the microplate is at least 5° and no greater than 20°.
47. The system of claim 45, wherein the means for inclining the microplate is fixed at a pre-determined angle of inclination, or wherein the means for inclining the microplate provides a single predetermined angle of inclination when in use; or wherein the means for inclining the microplate is adjustable, so that the microplate may be moved from a substantially horizontal position to a desired angle of inclination.
48. The system of claim 47, wherein the means for inclining the microplate comprises a support of adjustable height which may engage with different parts of the microplate to produce a variable angle of inclination.
49. The system of claim 45, wherein the means of inclining the microplate comprises an inclined platform on which the microplate is to be located, or support such as one or more rotating legs, telescopic legs or piston arrangements.
50. The system of claim 45, further comprising a means for engaging the microplate, wherein an engaging means is one or more raised areas or protrusions on the microplate holder, which engage with the microplate to restrict a relative movement of the microplate and microplate holder, or one or more indentations or recesses on the microplate holder, which engage with the microplate to restrict a relative movement of the microplate and microplate holder.
51. The system of claim 45, further comprising a locking means to securely engage the microplate.
52. The system of claim 51, wherein the locking means is one or more securing clips located on the microplate holder.
53. The system of claim 45, further comprising a means of stabilizing the microplate holder.
54. The system of claim 53, wherein the means of stabilizing the holder comprises one or more handles.
55. An automated microplate device comprising a system according to claim 45, comprising a microplate washer head which can be raised and lowered and/or rotated for interaction with a microplate in an inclined position, wherein the microplate washer head comprises bespoke pairs of aspiration and dispending pins, which comprise a dispensing pin that directs fluid towards an internal wall of the microplate well.
Description
BRIEF DESCRIPTION OF THE FIGURES
[0190] FIG. 1A shows a cross-sectional view of a single well from a standard microplate in the conventional horizontal position, with a pipette tip introduced to aspirate or dispense solution.
[0191] FIG. 1B shows a plan view from above of a single well from a standard microplate in the conventional horizontal position.
[0192] FIG. 2A shows a cross-sectional view of a single well from a microplate with areas for communication with a pipette tip, and a guide channel, in accordance with an embodiment of the first aspect of the present invention, with a pipette tip introduced to aspirate or dispense solution.
[0193] FIG. 2B shows a plan view from above of the embodiment of FIG. 2A.
[0194] FIGS. 3A, 3B, and 3C show cross-sectional views of the effect of a pipette tip upon a monolayer of adherent cells which are growing on the bottom of a standard microplate well in the conventional horizontal position.
[0195] FIGS. 3D, 3E, and 3F show a plan view from above of the embodiments of FIGS. 3A, 3B, and 3C, respectively.
[0196] FIG. 4A shows a cross-section that demonstrates a microplate well wherein the area for communication with the pipette tip comprises two raised areas located on the base and wall of the well interior.
[0197] FIG. 4B shows a cross-section that demonstrates a microplate well wherein the area for communication with the pipette tip comprises an indentation located on the base of the well interior.
[0198] FIG. 4C shows a cross-section that demonstrates a microplate well wherein the area for communication with the pipette tip comprises a raised area located on the base of the well, and an indentation located on the wall of the well.
[0199] FIG. 4D shows a cross-section that demonstrates a microplate well wherein the area for communication with the pipette tip comprises indentations located on the base and the wall of the well interior.
[0200] FIG. 5A shows a cross-section of the areas for communication with the pipette tip comprise one indentation located on the base of the well interior, and tapered indentation in the wall of the well interior; and a guide channel, wherein the end of the guide channel that is located near the base of the microplate well terminates in the indentation in the well wall.
[0201] FIG. 5B shows a cross-section of a microplate well wherein the areas for communication with the pipette tip comprise one raised area located on the base of the well interior, and tapered indentation in the wall of the well interior; and a guide channel, wherein the end of the guide channel which is located near the base of the microplate well terminates in the indentation in the well wall.
[0202] FIG. 5C shows a cross-section of a microplate well wherein the areas for communication with the pipette tip comprise raised areas located on the base of the well interior, wherein one of the raised areas is located at the junction between the bottom of the well and the wall of the well, and a guide channel, wherein the end of the guide channel which is located near the base of the microwell terminates in a raised area.
[0203] FIG. 5D shows, in 3-dimensional form, preferred shapes for the raised areas for location on the base of the well interior.
[0204] FIGS. 5E and 5F show, in 3-dimensional form, preferred shapes for the indentation located on the base of the well interior.
[0205] FIG. 5G provides, in 3-dimensional form, an example of two raised areas for location within a microplate well, wherein one of the raised areas is located at the junction between the bottom of the well and the wall of the well.
[0206] FIG. 5H provides an alternative example of two raised areas for location within a microplate well in 3-dimensional form, wherein one of the raised areas is located at the junction between the bottom of the well and the wall of the well.
[0207] FIG. 5I shows, in three-dimensional form, two raised areas for location within a microplate, wherein one of the raised areas is located at the junction between the bottom of the well and the wall of the well, and the other is located on the base of the well; and a guide channel, wherein the guide channel terminates in an indentation in the wall of the well, which communicates with the raised area located at the junction between the bottom of the well and the wall of the well.
[0208] FIG. 5J shows, in three-dimensional form, two raised areas for location within a microplate, wherein one of the raised areas is located at the junction between the bottom of the well and the wall of the well; and the end of a guide channel, wherein the end of the guide channel terminates in the raised area located at the junction between the bottom of the well and the wall of the well.
[0209] FIG. 5K shows, in three-dimensional form, two raised areas located within a microplate on which the pipette tip can be located, a guide channel, and two additional raised areas located either side of the raised area which is located at the junction between the bottom of the well and the wall of the well. The additional raised areas prevent lateral movement of the pipette tip.
[0210] FIG. 5L shows, in three-dimensional and cross-sectional form, a microplate well wherein the area for communication with the pipette tip comprises an indentation in the wall of the well interior; and a guide channel, wherein the end of the guide channel which is located near the base of the microplate well terminates in the indentation in the well wall.
[0211] FIG. 5M shows, in three-dimensional and cross-sectional form, a microplate well wherein the area for communication with the pipette tip comprises an indentation in the wall of the well interior; and a guide channel, wherein the end of the guide channel which is located on the base of the microplate well terminates in the indentation in the well wall.
[0212] FIG. 5N shows, in three-dimensional and cross sectional form, an indentation in the interior well wall forming a guide channel, wherein the end of the guide channel terminates at the base of the well.
[0213] FIG. 5O, shows in three-dimensional form, a microplate well wherein the areas for communication with the pipette tip comprise two raised areas located on the base of the well interior, and an indentation in the wall of the well interior; and a guide channel, wherein the end of the guide channel which is located near the base of the microplate well terminates in the indentation in the well wall.
[0214] FIG. 6A shows, in three-dimensional form, two raised areas located on the interior wall of the microplate well forming a guide channel, wherein the end of the guide channel terminates in a raised area located at the junction between the bottom of the well and the wall of the well.
[0215] FIGS. 6B and 6C show, in 3-dimensional form, preferred shapes for the two raised areas forming a guide channel, wherein the end of the guide channel terminates in the raised area located at the junction between the bottom of the well and the wall of the well.
[0216] FIG. 6D shows, in 3-dimensional form, two raised areas located on the interior wall of the microplate well forming a guide channel, wherein the end of the guide channel terminates at the base of the well.
[0217] FIG. 7A shows a cross-sectional view and a plan view (from above) of an embodiment of a microplate in accordance with the present invention, wherein the area for communication with a pipette tip comprises two raised areas located on the wall and the base of the microplate well.
[0218] FIG. 7B shows a cross-sectional view and a plan view (from above) of an embodiment of a microplate in accordance with the present invention wherein the area for communication with a pipette tip comprises two indentations located on the wall and the base of the microplate well.
[0219] FIG. 8A shows a cross-sectional view and a plan view (from above) of an embodiment of a microplate in accordance with the present invention, wherein the areas for communication with the pipette tip comprise two indentations located on the base and wall of the well interior; and a guide channel, wherein the end of the guide channel which is located near the base of the microplate well terminates in the indentation in the well wall.
[0220] FIG. 8B shows a cross-sectional view and a plan view (from above) of an embodiment of a microplate in accordance with the present invention, wherein the areas for communication with the pipette tip comprise one raised area located on the base of the well interior, and an indentation in the wall of the well interior; and a guide channel, wherein the end of the guide channel which is located near the base of the microplate well terminates in the indentation in the well wall.
[0221] FIGS. 9A, 9B, and 9C show cross-sectional views of the effects of a pipette tip upon a monolayer of adherent cells which are growing on the bottom of a microplate well according to an embodiment of the first aspect of the present invention. FIGS. 9A, B and C show a cross-sectional view; FIGS. 9D, E, F show a plan view from above.
[0222] FIGS. 9D, 9E, and 9F show a plan view from above of the embodiments of FIGS. 9A, 9B, and 9C, respectively.
[0223] FIG. 10 shows a cross-section view of a single well from a standard microplate, held in an inclined position, with the pipette tip held in a position to aspirate or dispense solution.
[0224] FIGS. 11A, 11B, and 11C show a cross-sectional view of the effects of a pipette tip upon a monolayer of adherent cells in standard microplate well when the microplate is being held in an inclined position.
[0225] FIGS. 11D, 11E, and 11F show a plan view from above of the embodiments of FIGS. 211A, 11B, and 11c, respectively.
[0226] FIG. 12 shows a cross-sectional view of a microplate located on a microplate holder in accordance with the second aspect of the present invention, and a corresponding plan view of the microplate, as inclined.
[0227] FIG. 12A shows a 3-dimensional view of a microplate holder in accordance with an embodiment of the second aspect of the invention, comprising two handles, as a means for stabilizing the holder.
[0228] FIG. 13 shows a cross-section view of a microplate located on a microplate holderin accordance with the second aspect of the present invention.
[0229] FIG. 14A shows a cross-section view of a microplate located on a different microplate holder in accordance with the second aspect of the present invention.
[0230] FIGS. 14B and 14C provide details of a means for securing the microplate to the microplate holder, in accordance with the second aspect of the present invention.
[0231] FIG. 15 shows cross-section and plan views of a microplate located on a microplate holder in accordance with the second aspect of the present invention, as inclined.
[0232] FIGS. 15A, 15B, and 15C show alternative embodiments for inclining a microplate.
[0233] FIG. 16 shows cross-section and plan views of a microplate located on a different microplate holder in accordance with the second aspect of the present invention, as inclined.
[0234] FIG. 17 shows cross-section and plan views of a microplate located on a further microplate holder in accordance with the second aspect of the present invention, as inclined.
[0235] FIG. 18 shows cross-section and plan views of a microplate located in a further microplate holder in accordance with the second aspect of the present invention when used in an automated system, as inclined.
[0236] FIG. 18A show a cross-sectional view of an alternative washerhead for use in an automated system, comprising angled washer pins.
[0237] FIG. 19 shows cross-section and plan views of a microplate located in a further microplate holder in accordance with the second aspect of the present invention when used in an automated system, as inclined.
[0238] FIG. 20 shows a cross-sectional view of a microplate in an automated system in accordance with the third aspect of the present invention, and a corresponding plan view of the plate, as inclined.
[0239] FIG. 20A illustrates various ways in which a washer head in an automated system may be manipulated in order to communicate with an inclined microplate.
[0240] FIGS. 21A and 21B show a pair of washer pins as used on a standard washer head in an automated system.
[0241] FIGS. 22A, 22B, 22C, and 22D show pairs of bespoke washer pins in accordance with the second aspect of the present invention.
[0242] FIG. 23A shows a single well of a microplate held in an inclined position, the microplate well comprising a guide channel comprising an indentation, and a raised area for communication with a pipette tip, in accordance with the first aspect of the present invention.
[0243] FIG. 23B shows a cross-sectional view of a microplate, wherein each well comprises a guide channel comprising a raised area on the wall of the well interior, in accordance with a first aspect of the present invention. The microplate is located on a microplate holder in accordance with the second aspect of the present invention.
[0244] FIG. 23C shows a cross-sectional view of a microplate, wherein each well comprises a guide channel comprising an indent in the wall of the well interior, in accordance with a first aspect of the present invention. The microplate is located on a microplate holder in accordance with the second aspect of the present invention.
DETAILED DESCRIPTION OF THE FIGURES
[0245] In FIG. 1, a pipette tip 1 is introduced into a standard microplate well 2 to remove residual liquid 3.
[0246] In FIG. 2, a pipette tip 1 is introduced into a microplate well in accordance with an embodiment of the first aspect of the present invention 2′ to remove residual liquid 3.
[0247] The well has a tapered guide channel 4, wherein the end of the guide channel which is located near the base of the microplate well terminates in the indentation in the well wall 5. The areas for communication with the pipette tip comprise the indentation located on the wall of the well interior 5, and the raised area located on the base of the well, 6.
[0248] In FIG. 3A, a pipette tip 1 is introduced to a microplate well 2 containing a monolayer of adherent cells 4, to remove the residual fluid 3.
[0249] In FIGS. 3B and 3C, the pipette tip contacts the base of well, scraping and dislodging the cells, resulting in removal of some of the cells from an area of the base of the well 5.
[0250] In addition, some of the cells are dislodged from the base of the well, but remain attached to cells which are affixed to the base of the well 6. These cells could detach from the well upon aspiration.
[0251] A demonstration of the cell distribution before introduction of the pipette tip is provided by FIG. 3D. A demonstration of the damage that can be caused to the cells by the introduction of the pipette tip is provided by FIGS. 3E and 3F.
[0252] An illustration of the cell distribution before introduction of the pipette tip is provided by FIG. 3D. An illustration of the damage that can be caused to the cells by the introduction of the pipette tip is provided by FIGS. 3E and F.
[0253] In FIG. 4A, a microplate well in accordance with an embodiment of the first aspect of the present invention 2′ has two raised areas 6. One raised area is located on the well wall, and the other is located on the well base. The raised areas are spaced so that when the pipette tip 1 is introduced, the edges of the pipette tip rest on the raised areas, so that a channel is formed between the pipette tip and the raised areas for the release and aspiration of fluid.
[0254] FIG. 4B shows a microplate well in accordance with an embodiment of the first aspect of the present invention 2′ with an indentation 5 located on the well base. An edge of the pipette tip 1 can be located within the indentation 5, whilst the other edge of the pipette tip rests against the well wall.
[0255] FIG. 4C shows a microplate well in accordance with an embodiment of the first aspect of the present invention 2′ with one raised area 6 located on the well base and one indentation 5 located on the well wall. The raised area and indentation are spaced so that when the pipette tip 1 is introduced, the edges of the pipette tip rest on the raised area 6 and in the indentation 5, so that a channel is formed between the pipette tip 1 and the areas for communication with the tip of a pipette 5 and 6, for the release and aspiration of fluid.
[0256] FIG. 4D shows a microplate well in accordance with an embodiment of the first aspect of the present invention 2′ with two indentations 5 located on the well base and wall. The indentations are spaced so that the pipette tip 1 can be located within the indentations, thereby forming a channel between the pipette tip 1 and the areas for communication with the tip of a pipette, for the release and aspiration of fluid.
[0257] FIG. 5A shows a microplate well in accordance with an embodiment of the first aspect of the present invention 2′ with a guide channel 4 comprising a tapered indentation in the interior wall of the well, wherein the end of the guide channel that is located near the base of the microplate well terminates in an indentation 5 which acts as an area for communication with the pipette tip 1; and a further indentation 5 located on the base of the well, so that the pipette tip 1 can be introduced to the well via the guide channel 4, and located in the two indentations 5 on the wall and base of the well, thereby forming a channel between the pipette tip 1 and the areas for communication with the tip of a pipette, for the release and aspiration of fluid.
[0258] FIG. 5B shows a microplate well 2′ in accordance with an embodiment of the first aspect of the present invention, designed, in particular, to prevent lateral movement of the pipette tip once located for aspiration or dispensing of fluid. The well comprises a guide channel 4 comprising a tapered indentation in the interior wall of the well, wherein the end of the guide channel that is located near the base of the microplate well terminates in an indentation 5 which acts as an area for communication with the pipette tip 1; and a raised area 6 located on the base of the well, so that the pipette tip 1 can be introduced to the well via the guide channel 4, and located in the indentation 5 on the wall of the well, and the raised area 6 on the base of the well, thereby forming a channel between the pipette tip 1 and the areas for communication with the tip of a pipette, for the release and aspiration of fluid.
[0259] FIG. 5C shows a microplate well in accordance with an embodiment of the first aspect of the present invention 2′ with a guide channel 4 comprising a tapered indentation in the interior wall of the well, wherein the end of the guide channel that is located near the base of the microplate well terminates in a raised area 6′ which acts as an area for communication with the pipette tip 1, and a further raised area 6 located on the base of the well, so that the pipette tip 1 can be introduced to the well via the guide channel 4, and located on the raised areas 6′ and 6, thereby forming a channel between the pipette tip 1 and the areas for communication with the tip of a pipette for the release and aspiration of fluid.
[0260] FIG. 5D shows, in three dimensional form, two preferred approximately pyramidal shapes for the raised areas 6 for location on the base of the well interior which can act as an area for communication with a pipette tip 1. FIGS. 5E and F show, in three dimensional form, preferred, approximately pyramidal shapes for the indentation 5 for location on the base of the well interior, which can act as an area for communication with a pipette tip 1.
[0261] FIG. 5G provides a three-dimensional depiction of the interior surface of a microplate well in accordance with a first aspect of the present invention 2″ with two raised areas 6′ and 6 located in the well interior, wherein one raised area 6′ is located at the junction between the bottom of the well and the wall of the well, and the other 6 is located on the base of the well interior, wherein the raised areas act as areas for communication with a pipette tip 1 and form a channel between the pipette tip 1 and the raised areas 6′ and 6 for the release and aspiration of fluid.
[0262] FIG. 5H provides an alternative three-dimensional depiction of the interior surface of a microplate well in accordance with a first aspect of the present invention 2″ with two raised areas 6′ and 6 located in the well interior, wherein one raised area 6′ is located at the junction between the bottom of the well and the wall of the well, and the other 6 is located on the base of the well interior, wherein the raised areas act as areas for communication with a pipette tip 1 and form a channel between the pipette tip 1 and the raised areas 6′ and 6 for the release and aspiration of fluid.
[0263] FIG. 5I provides a three-dimensional depiction of the interior surface of a microplate well in accordance with a first aspect of the present invention 2″ with a guide channel 4 comprising a tapered indentation in the interior wall of the well, wherein the end of the guide channel that is located near the base of the microplate well terminates in an indentation 5 which communicates with a raised area 6′, which is located at the junction between the bottom of the well and the wall of the well; a second raised area 6 is located on the base of the well interior. The pipette tip 1 can be introduced to the well via the guide channel 4, and located on the indentation 5 on the wall of the well and the raised area 6, or on the two raised areas 6′ and 6, in either case forming a channel between the pipette tip 1 and the areas for communication with the tip of a pipette, for the release and aspiration of fluid. At least partial location of the pipette tip in the indentation 5 in the wall of the well prevents lateral movement of the pipette tip.
[0264] FIG. 5J provides a three-dimensional depiction of the interior surface of a microplate well in accordance with a first aspect of the present invention 2″ with a guide channel 4 comprising a tapered indentation in the interior wall of the well, wherein the end of the guide channel that is located near the base of the microplate well terminates in a raised area 6′, which is located at the junction between the bottom of the well and the wall of the well with a second raised area 6 located on the base of the well interior. The pipette tip 1 can be introduced to the well via the guide channel 4, and located on the two raised areas 6′ and 6, forming a channel between the pipette tip 1 and the areas for communication with the tip of a pipette, for the release and aspiration of fluid.
[0265] FIG. 5K provides a three-dimensional depiction of the interior surface of a microplate well in accordance with a first aspect of the present invention 2″ with a guide channel 4 comprising a tapered indentation in the interior wall of the well, wherein the end of the guide channel that is located near the base of the microplate well terminates in a raised area 6′, which is located at the junction between the bottom of the well and the wall of the well with a second raised area 6 located on the base of the well interior, and two additional raised areas 6′″, located either side of the raised area which is located at the junction between the bottom of the well and the wall of the well 6′. The pipette tip 1 can be introduced to the well via the guide channel 4, and located on the two raised areas 6′ and 6, forming a channel between the pipette tip 1 and the areas for communication with the tip of a pipette, for the release and aspiration of fluid. The additional raised areas 6′″ prevent lateral movement of the pipette tip once it has been located on raised areas 6 and 6′.
[0266] FIG. 5L provides a three-dimensional and cross-sectional depiction of the interior surface of a microplate well in accordance with a first aspect of the present invention 2″ with a guide channel 4 comprising an indentation in the interior wall of the well, which is wider at the lip of the well, and narrower at the base of the well, wherein the end of the guide channel that is located near the base of the microplate well terminates in an indentation 5. The pipette tip 1 can be introduced to the well via the guide channel 4, and located on the indentation 5 on the wall of the well. The cross-sectional depiction shows a guide channel 4 that is of uniform depth into the well wall from the lip of the well until the point of communication with the pipette tip at indentation 5.
[0267] FIG. 5M provides a three-dimensional and cross-sectional depiction of the interior surface of a microplate well in accordance with a first aspect of the present invention 2″ with a guide channel 4 comprising an indentation in the interior wall of the well, which is wider at the lip of the well and narrower at the base of the well, wherein the end of the guide channel that is located on the base of the microplate well terminates in an indentation 5. The pipette tip 1 can be introduced to the well via the guide channel 4, and located on the indentation 5 on the wall of the well. The cross-sectional depiction shows a guide channel 4 that is of uniform depth into the well wall from the lip of the well until the point of communication with the pipette tip at indentation 5.
[0268] FIG. 5N provides a three-dimensional and cross-sectional depiction of the interior surface of a microplate well in accordance with the present invention 2″ with a guide channel 4 comprising an indentation in the interior wall of the well, which is wider at the lip of the well and narrower at the base of the well, wherein the end of the guide channel terminates on the base of the well. The pipette tip 1 can be introduced to the well via the guide channel 4, and located on the base of the microplate well. The cross-sectional depiction shows a guide channel 4 that is of uniform depth into the well wall from the lip of the well until the point of communication with the pipette tip.
[0269] FIG. 5O provides a three-dimensional depiction of the interior surface of a microplate well in accordance with the present invention 2″ with a guide channel 4 comprising an indentation in the interior wall of the well, which is wider at the lip of the well and narrower at the base of the well, wherein the end of the guide channel that is located near the base of the microplate well terminates in an indentation 5; raised areas 6 are located on the base of the well interior. The pipette tip 1 can be introduced to the well via the guide channel 4, and located on the indentation 5 on the wall of the well, or on the two raised areas 6, in either case forming a channel between the pipette tip 1 and the areas for communication with the tip of a pipette, for the release and aspiration of fluid. At least partial location of the pipette tip in the indentation 5 in the wall of the well, or on the two raised areas 6, prevents lateral movement of the pipette tip.
[0270] FIG. 6A provides a three-dimensional depiction of the interior surface of a microplate well in accordance with a first aspect of the present invention 2″ with a guide channel 4 formed by substantially parallel raised areas 6′″, wherein the end of the guide channel that is located near the base of the microplate well terminates in a raised area 6′, which is located at the junction between the bottom of the well and the wall of the well. The pipette tip 1 can be introduced to the well via the guide channel 4, and located on the raised areas 6′.
[0271] FIG. 6B provides an alternative three-dimensional depiction of the interior surface of a microplate well in accordance with a first aspect of the present invention 2″ with a guide channel 4 formed by raised areas 6′″ of the same cross-sectional area at the lip and base of the well but angled to the vertical to create a channel that is wider at the lip of the well than at the base, wherein the end of the guide channel that is located near the base of the microplate well terminates in a raised area 6′, which is located at the junction between the bottom of the well and the wall of the well. The pipette tip 1 can be introduced to the well via the guide channel 4, and located on the raised areas 6′.
[0272] FIG. 6C provides an alternative three-dimensional depiction of the interior surface of a microplate well in accordance with the present invention 2″ with a guide channel 4 formed by raised areas 6′″ of differing cross-sectional size, being smaller at the lip and larger at the base to create a channel that is wider at the lip of the well than at the base, wherein the end of the guide channel that is located near the base of the microplate well terminates in a raised area 6′, which is located at the junction between the bottom of the well and the wall of the well. The pipette tip 1 can be introduced to the well via the guide channel 4, and located on the raised areas 6′.
[0273] FIG. 6D provides a three-dimensional depiction of the interior surface of a microplate well in accordance with the present invention 2″ with a guide channel 4 formed by raised areas 6′″, wherein the end of the guide channel that is located near the base of the microplate well terminates at the base of the microplate well. The pipette tip 1 can be introduced to the well via the guide channel 4, and located on the base of the microplate well.
[0274] FIG. 7A shows a cross-sectional view and a plan view (from above) of a microplate well in accordance with an embodiment according to a first aspect of the present invention 2′, wherein the area for communication with a pipette tip comprises two raised areas 6 located on the wall and the base of the microplate well. The raised areas are spaced so that when the pipette tip 1 is introduced, the edges of the pipette tip rest on the raised areas, so that a channel is formed between the pipette tip and the raised areas for the release and aspiration of fluid.
[0275] FIG. 7B shows a cross-sectional view and a plan view (from above) of a microplate well in accordance with an embodiment according to a first aspect of the present invention 2′, wherein the area for communication with a pipette tip comprises two indentations located on the wall and the base of the microplate well. The indentations are spaced so that when the pipette tip 1 is introduced, the edges of the pipette tip rest in the indentations, so that a channel is formed between the pipette tip and the indentations for the release and aspiration of fluid.
[0276] FIG. 8A shows a cross-sectional view and a plan view (from above) of a microplate well 2′ in accordance with an embodiment according to a first aspect of the present invention with a guide channel 4 comprising a tapered indentation in the interior wall of the well, wherein the end of the guide channel that is located near the base of the microplate well terminates in an indentation 5 which acts as an area for communication with the pipette tip 1; and a further indentation 5 located on the base of the well, so that the pipette tip 1 can be introduced to the well via the guide channel 4, and located in the two indentations 5 on the wall and base of the well, thereby forming a channel between the pipette tip 1 and the areas for communication with the tip of a pipette, for the release and aspiration of fluid.
[0277] FIG. 8B shows a cross-sectional view and a plan view (from above) of a microplate well 2′ in accordance with an embodiment according to a first aspect of the present invention with a guide channel 4 comprising a tapered indentation in the interior wall of the well, wherein the end of the guide channel that is located near the base of the microplate well terminates in an indentation 5 which acts as an area for communication with the pipette tip 1; and a raised area 6 located on the base of the well, so that the pipette tip 1 can be introduced to the well via the guide channel 4, and located in the indentation 5 on the wall of the well, and the raised area 6 on the base of the well, thereby forming a channel between the pipette tip 1 and the areas for communication with the tip of a pipette, for the release and aspiration of fluid.
[0278] In FIGS. 9A-C, a pipette tip 1 is introduced to a microplate well 2′, in accordance with a first aspect of the present invention which contains a monolayer of adherent cells 7, via a guide channel 4, which terminates in an indentation 5. The well has a raised area 6 located on the well base. The indentation 5 and the raised area 6 are spaced so that the edges of the pipette tip 1 can be located in the indentation 5 and on the raised area 6. Contact with the well base is, therefore, avoided.
[0279] As shown in FIGS. 9D-F, the number of cells are disrupted by the introduction of the pipette 1, either resulting in complete removal from the well 8′, or partial dislodging of the cells which then remain attached to cells which remain affixed to the base of the well 9′ is minimized.
[0280] FIG. 10 demonstrates the effect of holding the microplate in an inclined position. The pipette is able to extract the fluid more easily and completely as the tip can be more readily positioned in the most appropriate part of the well, into which the fluid is being encouraged to flow.
[0281] FIG. 11A demonstrates the effect of holding the microplate in an inclined position. In FIGS. 11A, 11B and 11C, the pipette tip contacts the base of well. However, as the pipette tip is directed to a specific area of the well by the inclination of the microplate, there is reduced and more localised scraping and dislodging of cells, resulting in removal of fewer cells from an area of the base of the well 5. In addition, fewer cells 6 are dislodged from the base of the well and are potentially lost upon aspiration.
[0282] A demonstration of the cell distribution before introduction of the pipette tip is provided by FIG. 11D. A demonstration of the reduced damage that can be caused to the cells by the introduction of the pipette tip when the microplate is held in an inclined position is provided by FIGS. 11E and 11F.
[0283] In FIG. 12, a pipette tip 1 is introduced into a well of a microplate 7, which is being held in an inclined position by a microplate holder in accordance with a second aspect of the present invention. The microplate holder comprises a base portion 8, a means for inclining the microplate in the form of an inclined platform 9, and a means for securing the microplate to the platform in the form of two raised outer edges 10 on the platform 9 which have an overhang 11, forming a groove into which the lip of the microplate fits, thereby preventing lateral or upward movement of the microplate. The microplate is slid into position on the platform by communication with the internal surfaces of the raised outer edges. The microplate further comprises holes into which securing pins, 12, which can be inserted once the microplate is in position, in order to prevent lateral movement of the plate.
[0284] FIG. 12A shows a microplate holder comprising an inclined platform 9, with handles 9′ located at the lower aspect on either side of the holder. The handles provide a means for the operator to stabilize the holder by placing a hand on or over one of the handles and exerting downward pressure.
[0285] In FIG. 13, the microplate 7 is being held in an inclined position by a microplate holder in accordance with a second aspect of the present invention. The microplate holder comprises a means for securing the microplate to the microplate holder in the form of a spring-loaded clip 12′. Pressure on the clip lever 15 by the user in the direction of the arrow causes extension of the spring 13 to which the clip is attached, causing the clip to open and allowing the microplate to be placed in the holder and, when the clip is returned to its rest position, it holds the microplate in place. The microplate holder may be located directly on a surface such as a laboratory bench 14, by resting one edge of the inclined platform on this surface.
[0286] FIG. 14A shows alternative means for securing the microplate to the microplate holder, in the form of a rotating clip 16, into which one edge of the microplate can be inserted. Once the microplate is thus engaged with the microplate holder, the microplate may be moved to an inclined position, with a further edge of the microplate being held in position by a slider or spring loaded clip 12.
[0287] FIG. 14B shows, in 3-dimensional form, further details of the rotating clip shown in FIG. 14A. The cylinder 16 comprises an opening 16′ into which one edge of the microplate can be inserted. The cylinder can then be rotated downwards, to allow the microplate to be moved into an inclined position. Two pins 17 interact with an aspect of the microplate holder in order to restrict the movement of the cylinder between two positions: a raised position which allows insertion of an edge of the microplate, and a lowered position which places the microplate at the required angle inclination. A handle, 16″ allows easier movement of the cylinder by the operator.
[0288] FIG. 14C shows, in 3-dimensional form, an alternative means for restricting the movement of a cylindrical rotating clip, comprising a shaped guide which forms part of the microplate holder. The cylinder is located in the guide, and the cylinder and microplate, once inserted, communicate with the guide at the extremes of rotation 17′, thus preventing free rotation of the cylinder.
[0289] In FIG. 15, the base portion of the microplate holder has a raised area or protrusion 21. One end of the microplate 7 is located over the raised area 21. This secures the microplate in one position and also prevents lateral movement.
[0290] FIG. 15A shows an alternative means for inclining the microplate, comprising a leg comprising indentations 18, into which an edge of the microplate 7 can be located.
[0291] FIGS. 15B and 15C show alternative means for inclining the microplate. FIG. 15B shows a screw, 19, which inserts into the underside of a platform of a microplate holder according to the present invention 9, and which can be adjusted using the nut, 20, which can be located on a surface such as a lab bench once adjustment is complete. When the nut is twisted, it engages or disengages the end of screw 19 from the platform 9, thus raising or lowering one end of the microplate.
[0292] FIG. 15C shows a screw 19, which has one end located in the base portion 8 of a microplate holder in accordance with the present invention, and an opposite end onto which the underside of microplate 7 can be located. The height of the screw 19 can be adjusted using the nut 20 which, when twisted, engages or disengages the end of screw 19 from the base portion 8 thus raising or lowering the end of the microplate 7.
[0293] FIG. 16 shows a further alternative embodiment of a microplate holder according to a second aspect of the present invention. A raised area 21 located on the base portion 8 of the microplate holder protrudes into the base of the microplate 7. An additional feature is a further raised area 22 on the microplate holder. This ensures the microplate does not slip forward.
[0294] FIG. 17 shows a further alternative embodiment of a microplate holder according to a second aspect of the present invention. A raised area 21 located on the base portion 8 of the microplate holder protrudes into the base of the microplate 7, and the holder comprises a further raised area 22. In addition, the microplate holder comprises an indentation 23 in which an edge of the microplate 7 is placed. This provides stability, and restricts of movement by the microplate 7.
[0295] FIG. 18 shows an embodiment of a microplate holder according to the present invention in an automated system. A microplate holder 23 maintains the microplate 7 at a fixed angle, x. The microplate holder comprises an indentation 24 into which the base of the microplate is located. The automated microplate washer head 25 has pairs of pipette tips 1, each pair comprising two pipette tips of differing lengths, tailored for use with the present invention. The length of the pairs of pipette tips decreases across the microplate, to compensate for the angle the microplate is held at by the microplate holder. The microplate washer head 25 remains horizontal, and moves in a lateral and/or vertical direction, so that it is located over, and raised and lowered over the inclined plate to allow aspiration and dispensing of fluid by the pipette tips.
[0296] FIG. 18A shows an alternative washer head for use in an automated system. A microplate holder according to a first aspect of the present invention 23 maintains the microplate 7 at a fixed angle, x. The microplate holder comprises an indentation 24 into which the base of the microplate is located. The washer head 25 has pairs of angled pipette tips or pins 1, each pair comprising two pipette tips of differing lengths. The length of the pairs of pipette tips decreases across the microplate. The microplate washer head 25 remains horizontal, but is capable of lateral movement so that it can be positioned accurately over the inclined plate. The washer head is also capable of vertical movement (either perpendicularly to the horizontal, or at an angle to the horizontal so that the upward and downward motion is ‘diagonal’), so that it can be raised and lowered in order to locate the pipette tips in the microplate wells, in order to effect aspiration from and release of fluid to the wells.
[0297] FIG. 19 demonstrates a further variation in the washer head in an automated system. A microplate holder 23 maintains the microplate 7 at a fixed angle, x. The microplate holder comprises an indentation 24 into which the base of the microplate is located. The automated microplate washer head 25 has pairs of pipette tips 1, each pair comprising two pipette tips of differing lengths for aspirating and dispensing fluid. The microplate washer head 25 is rotated to an angle which is substantially the same as the angle of inclination of the microplate by a pin 26, in order to compensate for the angle that the microplate is held at by the microplate holder. The microplate washer head 25 also moves in a vertical direction, so that it is raised and lowered over the inclined plate to aspirate and dispense fluid.
[0298] FIG. 20 shows an embodiment of a device according to the third aspect of the present invention. The device comprises a plate 27 which makes up a conveyor, one end of which is raised or lowered by piston-driven arms 28 to the required angle x for aspiration and dispensing of fluid by the pipette tips. The microplate washer head 25 is rotated to the required angle about pin 26. The washer head is also capable of lateral and/or vertical movement, so that it can be positioned accurately over the inclined plate, and lowered and raised (either perpendicularly to the horizontal; or at an angle to the horizontal so that the upward and downward motion is ‘diagonal’) over the microplate in order to aspirate and dispense fluid.
[0299] FIG. 20A illustrates the ways in which a washer head in an automated system may be manipulated in order to communicate with a microplate inclined to the desired angle, x by a device according to a third aspect of the invention. In a, the washer head is lowered down a vertical axis, and then rotated into position to communicate with the microplate. In b, the washer head moves horizontally until it is located over the microplate, rotated to the required angle, and is then lowered down a diagonal axis to communicate with the microplate. In c, the washer head is lowered vertically, rotated to the required angle and then moved down a diagonal axis to communicate with the microplate. In d, the washer head is lowered down a vertical axis, and during this motion, it is rotated to allow it to communicate with the inclined microplate.
[0300] FIG. 21A shows a pair of washer pins as used on a standard washer head in an automated system, comprising a dispensing tip 30 and an aspirating tip 29, which are arranged for insertion into a microplate well, 2, to aspirate fluid located therein 3.
[0301] FIG. 21B shows the arrangement of a pair of washer pins as used on a standard washer head when the microplate is inclined.
[0302] FIGS. 22A to 22D show pairs of bespoke washer pins in accordance with a second aspect of the present invention. In FIGS. 22A and B, the dispensing tip 30 and aspirating tip 29 are angled away from each other. In FIGS. 22B-D, the dispensing tip 30 comprises a bend or ‘kink’, so that the end of the tip is directed towards the internal wall of the microplate well.
[0303] FIG. 23 demonstrates the effect of holding a microplate in accordance with the first aspect of the present invention in an inclined position, in order to provide a method for improved introduction or removal of fluid from a microplate well in accordance with the sixth aspect of the present invention. In FIG. 23A, a pipette tip 1 is introduced to a microplate well 2 which contains a monolayer of adherent cells 7, via a guide channel 4, which terminates in an indentation 5, in accordance with a first aspect of the present invention. The well has a raised area 6 located on the well base. The indentation 5 and the raised area 6 are spaced so that the edges of the pipette tip 1 can be located in the indentation 5 and on the raised area 6. Contact with the well base is, therefore, avoided.
[0304] In FIG. 23B, a pipette tip 1 is introduced into a well of a microplate 2 via a raised guide channel 13 in accordance with the first aspect of the present invention, which is being held, at a desired angle, x, by a microplate holder in accordance with a second aspect of the present invention. The microplate holder comprises a base portion 8, a means for inclining the microplate in the form of an inclined platform 9, and a means for securing the microplate to the platform in the form of two raised outer edges 10 on the platform 9 which have an overhang 11, forming a groove into which the lip of the microplate fits, thereby preventing lateral or upward movement of the microplate. The microplate is slid into position on the platform by communication with the internal surfaces of the raised outer edges. The microplate further comprises holes into which securing pins, 12, which can be inserted once the microplate is in position, in order to prevent lateral movement of the plate.
[0305] In FIG. 23C, a pipette tip 1 is introduced into a well of a microplate 2 via a indented guide channel 14 in accordance with the first aspect of the present invention, which is being held at a desired angle, x, by a microplate holder in accordance with a second aspect of the present invention. The microplate holder comprises a base portion 8, a means for inclining the microplate in the form of an inclined platform 9, and a means for securing the microplate to the platform in the form of two raised outer edges 10 on the platform 9 which have an overhang 11, forming a groove into which the lip of the microplate fits, thereby preventing lateral or upward movement of the microplate. The microplate is slid into position on the platform by communication with the internal surfaces of the raised outer edges. The microplate further comprises holes into which securing pins, 12, which can be inserted once the microplate is in position, in order to prevent lateral movement of the plate.
