Abstract
A pipette tip extension attachable to a pipette tip with the pipette tip extension having a proximal end, a distal end, and exterior wall extending between the proximal end and the distal end is disclosed. The exterior wall having an outer side and inner side and forming at the proximal end a reception aperture for inserting a pipette tip, and at a distal end a dispense aperture. An inner cavity is enclosed by the inner side of the exterior wall and one or more distance elements connected to the inner side of the exterior wall. The distance element(s) being configured to position a pipette tip within the inner cavity and to establish fluid uptake area adjacent the inner side of the exterior wall with the fluid uptake area extending from the dispense aperture towards the reception aperture and being in fluid connection with the surrounding atmosphere at the reception aperture.
Claims
1. A pipette tip extension (1) attachable to a pipette tip (20), the pipette tip extension (1) comprising: a proximal end (2), a distal end (3), and an exterior wall (4) extending between the proximal end (2) and the distal end (3), the exterior wall (4) having an outer side (5) and an inner side (6) and forming at the proximal end (2) a reception aperture (7) for inserting a pipette tip (14), and at the distal end a dispense aperture (8), an inner cavity (9) enclosed by the inner side (6) of the exterior wall (4), one or more distance elements (10) connected to the inner side (6) of the exterior wall (4), the distance element(s) (10) being configured to position a pipette tip (20) within the inner cavity (9) and to establish a fluid uptake area (11) adjacent to the inner side (6) of the exterior wall (4), the fluid uptake area (11) extending from the dispense aperture (8) towards the reception aperture (7) and being in fluid connection with the surrounding atmosphere at the reception aperture (7), the pipette tip extension being configured as an integrally produced workpiece, and being made of a non-polar olefine based polypropylene (PP) composition comprising at least 85% of a heterophasic polypropylene copolymer (h-PP), the latter comprising a propylene random copolymer matrix phase (r-PPM), an elastomeric propylene copolymer (e-PP) dispersed in the matrix, and optionally the PP composition may comprise polyethylene (PE) dispersed in the matrix, whereat the polypropylene composition has a shore hardness D from 45-60.
2. The pipette tip extension according to claim 1, whereat the polypropylene composition has: a melting temperature between 130-160° C. and a melt flow rate (MFR, 230° C./2.16 kg) according to ISO 1133 from 0.5-6.0 g/10 min.
3. The pipette tip extension according to claim 1, whereat the heterophasic polypropylene copolymer (h-PP) comprises at least 65 to 90 wt-% of the propylene random copolymer matrix phase (r-PPM), whereat the comonomer of the r-PPM is ethylene and/or C.sub.4 to C.sub.12 olefins, and the h-PP comprises 15 to 35 wt-% of a dispersed phase comprising an elastomeric propylene copolymer (e-PP) having ethylene and/or C.sub.4 to C.sub.12 olefins as comonomers.
4. The pipette tip extension according to claim 1, whereat the r-PPM has ethylene as comonomer and/or the e-PP has ethylene as comonomer.
5. The pipette tip extension according to claim 4, whereat the ethylene comonomer content of the r-PPM is 3 to 20 wt-% with reference to the r-PPM.
6. The pipette tip extension according to claim 4, whereat the ethylene comonomer content of the e-PP is 20 to 40 wt-% with reference to the e-PP.
7. The pipette tip extension according to claim 1, whereat the content of the PE dispersed in the matrix phase is equal or less than 15 wt-% of the composition.
8. The pipette tip extension according to claim 1, the composition further comprising a polymeric α-nucleating agent in a range of 0.0001 to 5 wt-% of the composition.
9. The pipette tip extension according to claim 1, further comprising a constriction element (12) to control an insertion depth of a pipette tip (20) within the pipette tip extension (1), the constriction element defining a gap height between a distal end of an inserted pipette tip and the distal end of the pipette tip extension.
10. The pipette tip extension according to claim 9, the constriction element being configured as a fluid-permeable sieve-like structure, wherein the constriction element (12) comprises a guiding plate (14), support bars (15) and a connection channel (16), wherein the guiding plate (14) defines or is connected to an end stop (23) for a pipette tip (20), the guiding plate (14) being connected to an inner side (6) of the exterior wall (4) by the support bars 15 close to but offset of a distal end (3) of the pipette tip extension (1), delimiting together with the inner side (6) of the exterior wall (4) a gap (17) of a gap height (h) at the distal end (3) of the pipette tip extension (1), the support bars (15) providing one or more passages for fluidly connecting the gap (17) with the fluid uptake area (11).
11. The pipette tip extension according to claim 1, wherein the distance element(s)(10) or the constriction element is or forms a sleeve (21) which is configured to receive and circumferentially enclose a distal end of a pipette tip (20) within the inner cavity (9), wherein the sleeve (21) is adapted to the shape of a distal end of a pipette tip (20) in a liquid-tight manner.
12. The pipette tip extension (1) according to claim 1, comprising at least two, preferably at least three additional distance elements (10), each distance element (10) being configured as a distance bar (24) which extends along a direction from the proximal end (2) towards the distal end (3) of the pipette tip extension (1), wherein the form of the distance bars and the position of the distance bars are configured such that each distance bar (24) can be abutted by an outer side of a pipette tip (20) when inserted into the pipette tip extension (1).
13. The pipette tip extension according to claim 9, wherein the distance element(s) (10) is connected to the inner side (6) of the exterior wall (4), and the constriction element (12) is connected to the inner side (6) of the exterior wall (4) of the pipette tip extension (1), thereby defining an end stop (23) for a pipette tip (20), and being configured as a fluid-permeable sieve-like structure.
14. The pipette tip extension (1) according to claim 1, wherein the distance element (10) is configured as a sleeve (21) for receiving and circumferentially enclosing a distal end of a pipette tip (2), the sleeve (21) being open at the distal end for allowing a liquid being dispensed from an inserted pipette tip (20) into the gap (17), and wherein the end stop (23) is configured as an elongated, tubular hollow cylinder which provides a passage for fluidly connecting an inserted pipette tip (20) with the gap (17), wherein the sleeve (21) and the end stop (23) are fluidly connected to provide the fluid connection between the pipette tip (20) and the gap (17).
15. The pipette tip extension (1) according to claim 1, wherein the guiding plate (14) is configured as a ring-like guiding plate (14) or as a shim-like guiding plate (14) or as a plate-like guiding plate (14).
16. The pipette tip extension according to claim 10, wherein the one or more support bars (15) connecting the guiding plate to the inner side (6) of the exterior wall (4), or a connection element (28) connecting the inner wall to the sleeve have a cut out section (29) provided to extend the height (h) of the gap immediately adjacent to the inner side (6) of the exterior wall (4).
17. The pipette tip extension according to claim 1, wherein the pipette tip extension is produced by injection molding of the respective polypropylene composition.
18. The pipette tip extension according to claim 1, wherein the parting line (30) of at least two mold parts used to produce the pipette tip extension is shifted into the dispense aperture (8) or into the connection channel (16).
Description
BRIEF DESCRIPTION OF THE DRAWINGS
[0119] Embodiments of the state of the art and the current invention are described in more detail in the following with reference to the figures (FIG.). As parts from WO2020/13232394 represent an integral part of the present invention FIGS. 1A to 7 are overtaken from there to explain respective mechanical and geometrical features which are important to understand also for the present invention. The figures following thereafter represent certain further developments and experiments and further features of improvement (figures x-y).
[0120] FIG. 1A a schematic perspective side view of a pipette tip extension in an exemplary embodiment,
[0121] FIG. 1B a schematic top view into a pipette tip extension of FIG. 1A,
[0122] FIG. 1C a schematic sectional drawing of the pipette tip extension of FIG. 1A based on intersection plane C-C,
[0123] FIG. 1D a schematic sectional drawing of the pipette tip extension of FIG. 1A based on intersection plane D-D,
[0124] FIG. 2A a schematic bottom view drawing onto the distal end of a pipette tip extension of FIG. 1A,
[0125] FIG. 2B a schematic sectional drawing of an exemplary embodiment of the distal end of a pipette tip extension based on intersection plane A-A,
[0126] FIG. 3A a schematic, perspective drawing onto the distal region of a pipette tip extension in a further embodiment,
[0127] FIG. 3B a schematic, perspective drawing of a sectional view of the distal region of the pipette tip extension of FIG. 3A,
[0128] FIG. 3C a schematic, sectional drawing of the pipette tip extension of FIG. 3A with an inserted pipette tip,
[0129] FIG. 4A a schematic sectional drawing of the distal end of the pipette tip extension of FIG. 3A,
[0130] FIG. 4B the schematic sectional drawing of FIG. 4A with inserted pipette tip, the pipette tip extension being positioned onto a flat surface,
[0131] FIG. 5A a schematic perspective drawing of a view onto the distal region of a pipette tip extension in a further embodiment,
[0132] FIG. 5B a schematic perspective drawing of a view onto the proximal region of the pipette tip extension of FIG. 5A,
[0133] FIG. 6 a schematic perspective sectional drawing of the pipette tip extension of FIG. 5B based on intersection plane A-A,
[0134] FIG. 7 a schematic sectional drawing of the pipette tip extension of FIG. 5B based on intersection plane B-B,
[0135] FIG. 8A-D a comparative example of a two-part pipette tip extension,
[0136] FIG. 9A-C a comparative examples of a one-part pipette tip extension,
[0137] FIG. 10A-C comparative examples of one-part pipette tip extensions,
[0138] FIG. 11A-B a comparative example of a one-part pipette tip extension,
[0139] FIG. 12A-B a comparative example of a one-part pipette tip extension,
[0140] FIG. 13A-B details of the distal end region of one-part pipette tip extensions,
[0141] FIG. 14A-C a one-part pipette tip extension according to the invention,
[0142] FIG. 15A-C a one-part pipette tip extension according to the invention
[0143] FIG. 16A-C a one-part pipette tip extension according to the invention.
DETAILED DESCRIPTION OF THE FIGURES
[0144] FIGS. 1A to ID show an exemplary embodiment of a pipette tip extension 1 in a perspective drawing. The pipette tip extension 1 here is an elongated, tube-like body with a proximal end 2 and a distal end 3. An exterior wall 4 extends between the proximal end 2 and the distal end 3, and forms at the proximal end 2 a reception aperture 7 and forms at the distal end 3 a dispense aperture 8. The exterior wall 4, comprising an outer side 5 and an inner side 6, encloses an inner cavity 9. The inner cavity 9 is delimited by the inner side 6 of the exterior wall 4. The medial axis 13 is indicated and corresponds here to the longitudinal axis of the pipette tip extension 1.
[0145] The pipette tip extension 1 shown here is a rotationally symmetric body. The reception aperture 7 is configured to receive a pipette tip 20 when the pipette tip extension 1 shall be attached to that pipette tip 20. The dimensions, in this case the diameter, of the reception aperture 7 is in particular bigger than the diameter of a pipette tip 20. The dispense aperture 8 is configured being abuttable to a flat surface in a sealing manner. The area of the flat surface, which is below the dispense aperture when a pipette tip extension 1 is sealingly abutted to that surface, and which is as a consequence also below the gap 17, may then be contacted by a liquid which is dispensed from a pipette tip 20 which is inserted into the pipette tip extension 1.
[0146] In the present context, the term “a pipette tip extension 1 attachable/attached to a pipette tip 20” describes essentially the same situation as the term “a pipette tip 20 insertable/inserted into a pipette tip extension 1”.
[0147] The pipette tip extension 1 shown in FIG. 1A comprises a more proximal part which is formed as a cylindrical tube and a more distal part which tapers conically towards the distal end 3. This shape is an example of adapting the shape of the pipette tip extension 1 to the shape of a pipette tip 20 which shall be inserted. An additional measure for adapting the pipette tip extension 1 to the shape of a pipette tip 20 is for example the configuration of the distance elements 10 used. In the embodiment shown if FIG. 1A, three distance elements are used which are configured as elongated bars and which protrude into the inner cavity 9 towards the medial axis 13. Each of the elongated bars comprise a surface 18 which faces towards the medial axis 13, and which is configured as being abuttable at that surface 18 by an outer side of a pipette tip 20. A pipette tip may in this way being positioned within the inner cavity 9.
[0148] FIG. 1B shows a schematic top view of the proximal end 2 and into an embodiment of a pipette tip extension 1 which is identical with the embodiment of the pipette tip extension of FIG. 1A, except that the distance elements 10 are not arranged flush with the proximal end 2 but are arranged in an offset to the proximal end 2, as it is shown also in the FIGS. 1C and 1D. The surface of the upper edge of the pipette tip extension 1 is visible and is formed in this embodiment by the proximal end of the exterior wall 4. The distance elements 10 are arranged on the inner side 6 of the exterior wall 4 and protrude into the inner cavity 9. The distance elements 10 thereby establish a fluid uptake area 11 which is adjacent to the inner side 6 of the exterior wall 4 and within the inner cavity 9. The dimensions of the distance elements 10 may in this embodiment have a direct influence onto the volume of the fluid uptake area 11.
[0149] In FIG. 1B, the course of the intersection planes C-C and D-D are indicated, on which the FIGS. 1C and 1D are based on.
[0150] In FIG. 1C, a schematic sectional view of the pipette tip extension 1 according to the intersection plane C-C is shown. The pipette tip extension 1 in an embodiment shown in FIG. 1B can be seen here in a longitudinal section, which particularly allows a view onto one of the distance elements 10 which is configured as an elongated distance bar. In contrast to the embodiment shown in FIG. 1A, the distance elements 10 are arranged with an offset to the proximal end 2, as indicated by respective lines in FIG. 1B. The inner bar extends over almost the entire length of the pipette tip extension 1. The surface 18 which is abuttable by a pipette tip 20 can be seen frontally. FIG. 1C also allows a view onto the distal end 3 of the pipette tip extension, in particular to the dispense aperture 8 and the constriction element 12. The constriction element 12 is arranged with an offset to the dispense aperture 8 and is comprises in this embodiment sieve-like support bars 15 which form together a ring-shaped constriction element, as can be seen in more detail in FIG. 2A. The distance bars for positioning a pipette tip 20 extends up to the constriction element 12. The constriction element 12, in particular the ring-like structure (compare e.g. FIG. 2A) may be directly abutted by a pipette tip 20 when the pipette tip 20 is inserted into the pipette tip extension 1. The ring-like structure here serves as an end stop 23 which restricts the insertion depth of a pipette tip 20, while the ring-like structure itself is fixed within the inner cavity 9 by the support bars 15. As the inner opening of the ring-like structure provides a fluid connection between the inner cavity of an inserted pipette tip 10 and the gap 17, and the interspaces between the support bars 15 provide a fluid connection between the gap 17 and the fluid uptake area 11 of the pipette tip extension 1, the constriction element 12 allows by the sieve-like structure a liquid being dispensed from an inserted pipette tip 20 and moved into the fluid uptake area 11 when the gap 17 is closed by a flat sample or a corresponding flat surface. The gap 17 may be seen in more detail in FIG. 2B, which shows the distal end 3 of the pipette tip extension 1 shown in FIG. 1C in a higher magnification.
[0151] In FIG. 1D, a schematic sectional view of the pipette tip extension 1 according to the intersection plane D-D is shown. This longitudinal section allows a frontal view of one of the distance bars being cut (left side) and one of the bars shown in a side view (right side). Visible here is also the connection channel 16, which is provided by the ring-shaped constriction element 12.
[0152] In FIG. 2A, a schematic bottom view drawing onto the distal end 3 of a pipette tip extension 1 is shown, the embodiment of the distal end 3 corresponds here to the embodiment of the distal end 3 of a pipette tip extension shown in FIG. 1A or FIG. 1C. In this view, the sieve-like structure of the constriction element 12 at the distal region of the pipette tip extension can be seen. The constriction element 21 comprises here three support bars 15 which are arranged star-like at the distal end 3 of the pipette tip extension 1 and which connect a ring-shaped structure, the guiding plate 14, to the exterior wall 4. The support bars 15 extend laterally with respect to the medial axis of the pipette tip extension 1, and form together with the ring-shaped guiding plate 14 the constriction element 12. The ring-like guiding plate 14 here functions as an end stop, as the guiding plate 14 is abuttable by the distal end of a pipette tip 20 and prevents that the pipette tip 20 is inserted further into the inner cavity 9 of the pipette tip extension 1 (compare to FIG. 2B). By the offset of the constriction element 12 to the sealing surface 19 at the distal end 3 of the pipette tip extension 1, as may be seen in FIG. 2B, a gap 17 of a gap height h is formed between the distal end 3 of the pipette tip extension 1 and the distal end of an inserted pipette tip 20 (compare again with FIG. 2B). The sieve-like configuration ensures that a liquid which is dispensed out of an inserted pipette tip 20 may flow into the gap 17, over a surface or a sample thereon, when the pipette tip extension 1 is sealingly positioned on such a surface, and into the fluid uptake area 11 within the inner cavity 9 of the pipette tip extension 1.
[0153] In FIG. 2A, the course of the intersection plane A-A is indicated, on which the FIG. 2B is based on.
[0154] In FIG. 2B, a schematic sectional drawing of an exemplary embodiment of the distal end of a pipette tip extension based on intersection plane A-A, indicated in FIG. 2A, is shown. In this longitudinal section, the formation of the gap 17 at the distal end of the pipette tip extension 1 and the gap height h can be seen in more detail. The constriction element 12 is abuttable by a distal end of a pipette tip 20, as discussed above. The gap 17 is formed at the distal end 3 of the pipette tip extension 1, when the pipette tip 20 comes to rest within the pipette tip extension 1 with an offset to the distal end 3 of said pipette tip extension 1. When a pipette tip extension 1 is placed for example onto a flat surface comprising a tissue section, the seal section 19 at the distal end 3 of the pipette tip extension 1 provides a seal, and the gap 17 is closed by the flat surface, or a flat sample, e.g. a tissue section placed thereon, respectively. In this situation, a fluid connection between an inserted pipette tip 20 and the fluid uptake area 11 in the inner cavity 9 of the pipette tip extension 1 is provided by the gap 17, while the sieve-like configuration of the constriction element 12 does not hinder a continuous fluid flow although it defines an end stop 23 for the pipette tip 20. As the fluid uptake area 11 is additionally in fluid connection with the surrounding atmosphere at the proximal end 2 of the pipette tip extension 1, it is ensured that a liquid may be moved between the interior of a pipette tip 20 and the fluid uptake area of the pipette tip extension when they are assembled to a functional unit. By means of the gap height h, a flow velocity of the fluid flow may be influenced.
[0155] In FIG. 3A, a schematic, perspective drawing of a pipette tip extension 1 in another embodiment is shown as an overview, with a particular view onto the distal end 3 of the pipette tip extension 1 and the sieve-like structure of the constriction element 12. Also in this embodiment, the dispense aperture 8 can be seen with the constriction element 12 being arranged offset of the distal end 3, thereby forming a gap 17 at the distal end 3 of the pipette tip extension 1 (compare e.g., FIG. 4A). The offset of the constriction element 12 from the distal end 3 of the pipette tip extension 1, in particular the offset of the end stop 23 defined by the constriction element 12, directly influences the gap height h of the gap 17, as discussed before.
[0156] The constriction element 12 comprises a guiding plate 14, which has in this embodiment more a shim-like shape, and a central connection channel 16 which provides a fluid connection from an inserted pipette tip 20 into the gap 17. The constriction element 12 here also comprises support bars 15 which connect the guiding plate 14 to the inner side 6 of the exterior wall 4. The support bars 15 leave here by the interspaces between the support bars 15 three fluid passages at the inner side of the exterior wall 4, which allow a fluid being moved between the gap 17 and the fluid uptake area 11. The combination of shim-like guiding plate 14, the connection channel 16 and the support bars 15 provide the fluid-permeable sieve-like structure of the constriction element, which allows a fluid passage from an inserted pipette tip 20 through the connection channel 16 into the gap 17 and further from the gap 17 into the fluid uptake area 11 (compare also with FIG. 4B). The more shim-like shape of the guiding plate 14 allows the pipette tip extension 1 being configured for the treatment of medium-sized sample areas, as it provides more lateral stability, for example, compared to the embodiment of the pipette tip extension 1 shown in the FIGS. 1A to 2B, which are particularly useful in the treatment of small sample areas, as discussed above. The seal section 19 at the distal end 3 which provides a sealing connection to a flat surface, for example comprising a sample, is indicated.
[0157] In FIG. 3B, a schematic, perspective drawing of a sectional view of the distal region of the pipette tip extension of FIG. 3A is shown. This longitudinal section allows a perspective side view onto the constriction element 12 and an additional view onto the distance element 10, which is configured here as a sleeve 21 and which joins the constriction element 12 towards the proximal end 2 of the pipette tip extension 1. The sleeve 21 is configured to enclose a distal end of a pipette tip 20 when it is inserted, as may be seen in FIG. 3C and in more detail in FIG. 4B, without hindering a fluid being moved of out or into the inner cavity of such an inserted pipette tip 20. The distal end of the sleeve 21 is directly followed (in this case adjoined) by a thickened guiding plate 14. The guiding plate 14 comprises a connection channel 16 which mouths into the distal end of the sleeve 21. The connection channel 16 has an upper (faced toward the proximal end 2) diameter which is slightly smaller than the diameter of the adjacent distal end of the sleeve, so that a shoulder is formed at the distal end of the sleeve 21.
[0158] This shoulder is abutted by a distal end of a pipette tip 20 which is inserted into the pipette tip extension 1 and thus functions as an end stop 23 for controlling an insertion depth of a pipette tip 20. A pipette tip 20 which abuts the shoulder is thereby prevented to further move towards the distal end 3 of the pipette tip extension 1. The connection channel 16 here comprises inner bores of different diameters, which may help in guiding a fluid with a controlled flow velocity from an inserted pipette tip 20 into the gap 17. The sleeve 21, the connection channel 16 and the guiding plate 14 are integrally formed here, and commonly provide a positioning function (by the sleeve 21), a control of the insertion depth of a pipette tip 20 (by the end stop 23) while providing a fluid connection from an inserted pipette tip 20 into the gap 17. The support bars 15, which connect the guiding plate 14 to the inner side 6 of the exterior wall 4 are visible here, as well as the open passages which are left between the support bars 15 at the inner side 6 of the pipette tip extension 1.
[0159] In the embodiment shown in FIG. 3B, the distance element 10 comprises in addition to the sleeve 21 distance bars 24 which are arranged at the inner side 6 of the exterior wall 4 and which protrude here with an offset from the proximal end 2 of the pipette tip extension 1 towards the distal end 3 (compare with FIG. 3C). The distance bars 24 may be configured analogously to the distance bars shown in FIGS. 1A to 2B. The distance bars 24 are configured here however to join the proximal end of the sleeve 21 (see for example the right side of the exterior wall 4, where a distance bar 24 is shown to be cut and to join the sleeve 21). The distance bars 24 here provide additional support in positioning a pipette tip 20 which is inserted into a pipette tip extension 1. These distance bars 24 are here particularly adapted to the outer shape of the inserted pipette tip 20 (see also FIG. 3C, left side of the longitudinal section), in that the extent, by which each distance bar 24 protrudes towards the medial axis 13 is adapted in such a way that each distance bar is abutted by the distal region of an inserted pipette tip 20. The distance bar 24 on the right side is shown being cut. The pipette tip extension 1 may comprise three distance bars 24 for example for supporting the positioning of an inserted pipette tip 20, though other numbers may be possible as well, as discussed above.
[0160] In FIG. 3C, a schematic overview drawing of a longitudinal section of the pipette tip extension 1 of FIG. 3A with an inserted pipette tip 20 is shown. In this drawing, a distance bar 24 on the left side is shown being cut. The arrangement of distance element 10 (the sleeve 21 in this case), constriction element 12 (guiding plate 14 and support bars 15) and connection channel 16 is shown in an overview, the details may be taken from FIG. 4B. In this longitudinal section, the fluid uptake area 11 at the inner side 6 of the pipette tip extension 1 is visible on the right side of the pipette tip extension, and the gap 17 at the distal end 3 is also visible, though FIG. 4B allows a more detailed view onto the gap 17 when a pipette tip 20 is inserted. The sleeve 21 is shown to hold the distal end of a pipette tip 20. The fluid passage which is allowed by the arrangement and configuration of the distance element 10 and the constriction element 12 can be seen in more detail in the FIG. 4A and in particular in the FIG. 4B, which are higher magnifications of the pipette tip extension 1 of FIGS. 3A-4C.
[0161] In FIG. 4A, the distal end 3 of an embodiment of a pipette tip extension 1 shown in FIGS. 3A-3C is presented in more detail in a schematic sectional drawing. In FIG. 4B, the same situation is shown but with a pipette tip 20 being positioned by the sleeve 21 and the distance bars 24, which also establish the fluid uptake area 11. The insertion depth of the pipette tip 20 is controlled by the end stop 23 of the constriction element.
[0162] In FIG. 4A, the gap 17 and the corresponding gap height h are shown in more detail. The gap 17 is restricted by the distal, lower surface of the guiding plate 14. As the guiding plate 14 is positioned with an offset from the distal end 3 of the pipette tip extension 1, the gap 17 is formed there. The gap 17 provides a fluid connection of the interior of the disposable pipette tip 20 and the fluid uptake area 11 in the inner cavity 9 of the pipette tip extension 1, wherein a connection channel 16 provides the fluid passage from an inserted pipette tip 20 (see FIG. 4B) to the gap 17, and further wherein the interspace between the outer edge of the guiding plate 14 and the inner side of the exterior wall 4 provides a fluid passage between the gap 17 and the fluid uptake area 11. The use and configuration of the support bars 15 ensure that the fluid connection is not interrupted by the guiding plate 14, and they contribute in the provision of the sieve-like openings, as well as the connection channel 16 arranged in the guiding plate 14. As the fluid uptake area 11 is additionally in fluid connection with the surrounding atmosphere at the proximal end 2 of the pipette tip extension 1, it is ensured that a liquid may be moved between the interior 18 of the pipette tip 20 and the fluid uptake area 11 of the pipette tip extension 1 when they are assembled. The distance bar 24 on the left side is shown to be cut.
[0163] In FIG. 4B, the same situation of FIG. 4A is shown but with an inserted pipette tip 20. The distal end of the inserted pipette tip 20 abuts the end stop 23. Furthermore, the functional assembly of pipette tip 20 and pipette tip extension 1 is placed onto a sample 27 on a microscopic slide 26. Exemplarily, the sample is a tissue section on the flat surface of a microscopic slide 26. The pipette tip extension 1 is sealingly attached onto the tissue section, and thereby restricts an area of interest which may be treated by liquid movements between the inserted pipette tip 20 and the pipette tip extension 1. The flow direction of a liquid which is dispensed here is indicated by dotted arrows. The fluid flow is enabled by the configuration and mutual arrangement of the distance element 10, configured here as a sleeve 21, the connection channel 16, the guiding plate 14 and the support bars 15. In particular the connection channel 16 in the guiding plate 14 and the support bars 15 contribute to the fluid-permeable, sieve-like structure of the constriction element
[0164] In FIG. 5A, a schematic perspective drawing of a view onto the distal end 3 of a pipette tip extension 1 in a further embodiment is shown. In this embodiment, the pipette tip extension 1 is adapted for treating a larger area of interest. For this, the dispense aperture 7 at the distal end 3 has a larger diameter compared to those of previously discussed embodiments. The guiding plate 14 provides a larger area for being contacted by a liquid which is dispensed through the connection channel 16 into the gap 17, though the gap height h may be the same or of a comparable size than the gap heights h in other embodiments.
[0165] Also in this embodiment, the guiding plate 14 is connected to the inner side 6 of the exterior wall 4 by support bars 15 which are configured to provide interspaces between the guiding plate 14 and the inner side of the exterior wall 4. These interspaces serve as fluid passages, so that the fluid-permeability by the sieve-like structure is given, and a fluid may be moved from the gap 17 into the fluid uptake area 11 and back. The guiding plate 14 is here however configured as having an even larger surface area which faces towards the gap 17 and which may be contacted by a fluid during use of the pipette tip extension, as the guiding plate here as a more plate-like shape. As a measure to ensure that as much of a liquid is moved back from the gap 17 into an inserted pipette tip 20 for example during an aspiration step, a groove 22 is provided on the surface of the guiding plate 14 which faces the gap 17. Due to the surface tension the groove 22 holds the liquid during aspiration until the complete circumference of the groove ring is reached. In this way, early air aspiration is avoided and the residual liquid left on the sample minimized. It is believed that the groove “attracts” a liquid, which is present on the guiding plate 14 more closely to the exterior wall 4, towards the connection channel 16.
[0166] In FIG. 5B, a schematic perspective drawing of a view onto the proximal region of the pipette tip extension of FIG. 5A is shown. It can be seen here that in contrast to the previously discussed embodiments the inner cavity 9 comprises additional substructures. Here, an additional inner wall 25 is present. The inner wall 25 is connected by distance bars 24 to the inner side 6 of the exterior wall 4 and connected to a distance element 10 which configured as a sleeve 21 by connection elements 28 (see FIGS. 6A and 6B). Furthermore, the inner wall 25 is connected with its distal end to the guiding plate 14, so that additional structural stability is provided. By use of such an inner wall 25, the volume of the fluid uptake area 11 may be influenced, as the position of the inner wall 25, in particular the distance to the inner side 6 of the exterior wall 4, may have a direct influence onto the volume of the fluid uptake area 11.
[0167] In FIG. 5B, the course of the intersection planes A-A and B-B are indicated, on which the FIGS. 6 and 7 are based on.
[0168] In FIG. 6, a schematic perspective sectional drawing of the pipette tip extension of FIG. 5B based on indicated intersection plane A-A is shown. The additional substructures for allowing the treatment of a larger area of interest while controlling the volume of the fluid uptake area 11 and thereby controlling the volume of required liquid for the treatment of a sample can be seen here in more detail. The sleeve 21 is arranged centrally, and the end stop 23, which is formed by a shoulder here at the immediate distal end of the hollow sleeve 21 and at the beginning of the connection channel 16, controls the insertion depth of a pipette tip 20 within the sleeve 21 and within the pipette tip extension 1 (compare FIG. 7). The sleeve 21, the end stop 23 and the guiding plate are integrally formed here, though they may also be formed separately, and being connected in alternative manners. A fluid from an inserted pipette tip 20 may be moved out of said pipette tip 20, into the connection channel 16 within the guiding plate 14, and further into the gap 17 below the lower surface of the large guiding plate 14.
[0169] The sleeve 21 is separated in this embodiment from the fluid uptake area 11 by an inner wall 25, which here extends circumferentially around the sleeve 21. The inner wall 25 further extends in a direction from the distal end 3 towards the proximal end 2. The inner wall 25 thereby restricts the fluid uptake area 11 towards the medial axis 13, and the dimension and position of the inner wall 25 may be used for example to further influence the volume of and the fluid flow within the fluid uptake area 11, as discussed before. The inner wall 25 here even prevents that the outer side of an inserted pipette tip 20 is contacted by the fluid which is used for the treatment of a sample.
[0170] The inner wall 25 is connected to the inner side 6 of the exterior wall 4 by distance bars 24. The distance bars 24 are here joined additionally with the support bars 15, though they may alternatively also be separate from each other, provided they do not restrict the fluid connection of the fluid uptake area 11 with the surrounding atmosphere at the proximal end 2 of the pipette tip extension 1. The inner wall 25 is further connected to the guiding plate 14, which provides additional stability here. In this perspective sectional view, the interspace which is formed between the support bars 15, the outer edge of the guiding plate 14 and the inner side 6 of the exterior wall 4 can be seen. As discussed also for the other embodiments of the pipette tip extension 1, these interspaces which are generated when for example support bars 15 are used, provide a fluid passage from the gap 17 into the fluid uptake area 11. Interspaces may alternatively be formed by more grid-like or even membrane like configurations of the constriction element, as long as they additionally provide the end stop function.
[0171] In FIG. 7, a schematic sectional drawing of the pipette tip extension of FIG. 5B based on intersection plane B-B is shown. In this longitudinal section, two of the connection elements 28 are cut. For a better overview, a pipette tip 20 is shown to be inserted into the sleeve 21. In this section it can be seen that the sleeve, the connection elements 28, the inner wall 25 as well as the end stop 23, the guiding plate 14, the support bars 15, the distance bars 14, and the exterior wall 4 are produced in one piece, for example by injection moulding technology.
[0172] Although the diameter of the pipette tip extension 1 is in this case relatively large (it may be for example about 15 mm), a relatively small gap height h may nevertheless be generated (for example 0.4 mm). A smaller gap height h is particularly advantageously when only a small amount of liquid shall be used, for example for increasing the concentration of material gained in an extraction reaction, which is carried out on a tissue section. A smaller gap may also be of an advantage for improving the effect of a temperature treatment, when such a temperature treatment is directed primarily to the sample, and which requires that the fluid, which is present in the gap 17 and onto the tempered sample, is adapted to the temperature of the sample. In a smaller gap 17, the liquid may adapt relatively fast to the temperature of the sample. By means of the inner wall, the volume of liquid which is moved between an inserted pipette tip 20 and the pipette tip extension 1 may be further influenced, for example by providing a relatively small fluid uptake area 11.
[0173] FIG. 8A to 8D show a two-part comparative example of a pipette tip extension, the body 1 of which may have most or all of the features of the one-part pipette tip extension as described in detail with FIGS. 3A to 4B. The body 1 as shown separately in a 3 D and a transparent 3D scheme in FIG. 8A may comprise a circumferential step 34 and can be made of any plastic being stiff enough to hold its geometry under pressure when pressed on a glass slide for dispense and aspiration operations. Additionally, elasticity should be high enough to allow a friction or form fit with the pipette tip 20 to be inserted. As shown in FIG. 8B to 8C, onto that body 1 a collar seal 31 is mounted, e.g., flash to a step 34 which acts as stop for the collar 31. The latter being dimensioned to protrude the distal end 3, for about some tenths of a mm when mounted, and being made of a soft material to provide a good sealing when the pipette tip extension is pressed onto the sample 27, respectively onto the surface of the slide 26. Materials used for the body can be plastics as mentioned above, however polypropylenes in general due to their high chemical inertness, see also below, and specifically PP homopolymers or only slightly blended PP polymers would be preferred due to the availability of high hardness grades, e.g., up to Shore hardness D of 90 and higher, and its high elasticity. A specific example of a PP homopolymer with tradename Borealis HF955MO, is given under C1 in table 1, below, where some comparative and inventive examples have been listed. The material of the collar seal can be any material being on the one side soft enough to perform a sealing function and on the other side stable enough to be easily mounted on the body. Such materials can be plastic or natural rubbers, e.g., a silicon-rubber with a high chemical inertness. Details of the geometry are shown in a 3D-type cross-section in FIG. 8C and a schematic cross-section in FIG. 8D. Only the principle set up of a pipette tip extension 1 having a distal end 3, which forms a gap 17 when set on a glass slide 26 between the slide and support bars 15, respectively guiding plate 14, as well as respective end stop 23, connection channel and fluid uptake area 11 are shown. Further details and function of respective pipette tip extensions can be taken from FIGS. 3A to 4B. Two-part pipette tip extensions as shown in FIG. 8A-D are perfectly fitted to perform reliable sealed dispense and aspiration operations and can be combined with a circumferential sleeve 21 to avoid shortcutting between the pipette tip and the fluid uptake area 11 as explained with FIG. 13B below.
[0174] However, as such pipette tip extensions to be used for mass analytical applications are expensive and not easily recyclable due to the two-part construction respectively due to two different materials to be used, intensive research efforts have been undertaken to develop a respective one-part, one material solution as shown in the following. An overview of some comparative and inventive examples is given in table 1, below.
[0175] A comparative example of a one-part geometry and the same material as the body of pipette tip extension in comparative example C1 is shown in FIGS. 9A to 9C as comparative example C2. Due to slight surface imperfections unavoidable due to the injection moulding process and the high hardness of the material, no reliable seal could be provided by pressing a pipette tip with a respective pipette tip extension mounted on the surface of a test slide, i.e., during dispense and/or aspiration operations the test fluid (usually lyses buffer) leaked between the seal section 19 of the pipette tip extension 1 and the glass slide 25.
[0176] FIG. 10A to 10C show specific geometry examples of further comparative examples C3, C4 and C6, all based on the same type of pipette tip extension 1 as shown in the transparent 3D scheme in FIG. 10A. Further details and variations with regards to such pipette tip extensions can be found in FIGS. 1A to 4B and respective description.
[0177] Contrary to C1 these types of pipette tip extensions comprise at least a constriction element 12, e.g., to provide an end stop for the pipette tip 23, which is not in use with embodiments as shown in FIGS. 9A to 9C of embodiment C2. Special embodiments of the distal end 3 region of pipette tip extensions according to embodiment C3, C4 and C6 are shown in FIGS. 10 A and 10B in schematic cross sections. Contrary to the pipette tip extension as shown in FIG. 9A, where three downwards tapering distance elements are provided from the very proximal end 2 until about a lower third of the pipette tip extension, the pipette tip extension as shown in FIG. 10A shows respective tapering distance elements 10 extending from an upper part of the pipette tip extension until the stop surface near the lower/distal end.
[0178] With embodiment C3 a pipette tip extension being configured according to FIG. 10B has been injection moulded from a relatively soft thermoplastic elastomer (TPE, tradename: Medipren OF601M) of a shore hardness A of 60 which provided good sealing properties. This material however proved to be not stable enough as the height h of gap 17 between the slide and the pipette tip extension varied strongly depending on the contact pressure which may vary depending on the type of liquid handling workstation and operation status (dispense, aspiration, idle).
[0179] Therefore, with comparative embodiment C4 a harder type of TPE, having a shore hardness A of 87 (tradename: Mediprene OF901M) has been tested with the same geometry, whereby a good sealing of the seal section 19 towards test slides 26 and less deformation could be achieved. Fluid short cuts occurred in isolated cases. The same results could be achieved with comparative example C5 where a pipette tip extension according to FIG. 8 (i.e., without a collar seal) and a respective type of TPE as with C4 have been used. However during PCR tests with the respective TPE material a substantial scatter in the test results has been observed, which might be attributed to leaching of unknown PCR inhibitors or detrimental high binding properties for DNA/RNA which have not been investigated in detail, as this TTP material has not been approved as a medical grad material up to now. Respective tests with geometries according to FIGS. 8A, here again without a collar seal, and 10B with the medical grade PP material used for comparative example C1 and C2 as described above, failed due to the same reasons as described with C2.
[0180] Therefore, further geometries according to comparative example C6 with constriction element 4, as shown in FIG. 10C, and comparative example C7 without constriction element as shown in FIGS. 11A and 11B, have been tested with a respective PP homopolymer material. FIG. 10C shows a pipette tip extension which differs to 10B only in details of the distal end 3 and the grade of magnification. The seal section 19 in FIG. 10C having a very reduced thickness of the exterior wall 4 at and near the distal end 3 to allow a higher pressure on the remaining contact surface to the glass slide and thereby allow mechanical deformation of the plastic to enforce sealing when the seal section 19 is pressed onto a plane surface. Similarly, the geometry of the external wall 4 has been varied in FIG. 11B at the seal section 19 of the pipette tip extension 1. Additionally, in this case an outer stop ring 33 has been integrally provided to the distal end 3, encompassing the seal section in a distance and some tenth of a millimetre recessed within, to provide mechanical stability after a certain degree of enforced deformation of the seal section 19 has been reached by pressing on the slide 26. However, both combinations of respective geometries and PP homopolymer material could not provide a consistently reliable sealing function of the distal end 3.
[0181] Comparative example C8 using a geometry as shown in FIGS. 12A and 12B, again without a constriction element, and using a liquid silicon rubber (trade name: Silpuran), having a shore hardness A of 70, has been tested to give a reliable seal between the slide and the pipette tip extension as well as holding the grade of deformation with reference to the respective geometry of gap 17 and fluid uptake area 11 tolerable. However also this solution did not prove to be very suitable for mass production as two relatively expensive components have to be injected simultaneously into a heated form to produce the fine structures of a pipette tip extension in a reliable and reproducible manner.
[0182] The seal itself can be, as far as an appropriate material is used for any one-part pipette tip extension, the seal section 19 of the respective distal end 3 itself, which thereby forms a sealing area, as shown in FIG. 2A, 3A, 3B, 4B, 7, 9C, 10B, 12B, 14C, 15C, 16B, or alternatively be a separate ring or collar gasket, like collar seal 31 as shown and described on the basis of FIG. 8B to 8C below.
[0183] In FIG. 13A a problem is shown which may occur with pipette tip extensions having a constriction element 12 and an end stop even in cases where a fluid tight sealing is provided between the seal section 19 or a separate seal 31 and the slide 26. So called fluid short cuts as symbolized by arrow 32 may occur between the pipette tip 20 and the constriction element 12, which can be the guiding plate 14. Thereby, only a part of the fluid volume dispensed from the pipette tip flows via the connection channel 16 into the gap 7 where the flow is redirected by the slide 26 towards the fluid uptake area 11, whereby sample 27 is flushed from the surface of the test slide during a dispense operation of the pipette. However, a part of the fluid volume may be lost for the flushing operation when a short cut 32 occurs. On the other hand, during aspiration operations when there is a leakage between the pipette tip 20 and the constriction element, again fluid or environmental air may be sucked in the opposite direction directly from the fluid uptake 11 instead of taking the intended flow path via the gap 17 and connection channel 16 back into the pipet, which again may result in an insufficient washing of the sample from the test slide.
[0184] In FIG. 13B a respective solution against shortcuts is shown which encompasses a pipette tip extension having a sleeve, where the distal end of the sleeve 21 is adjoined to a thickened guiding plate 14. The guiding plate 14 comprises the end stop 23, a connection channel 16 which mouths into the distal end of the sleeve 21. The connection channel 16 has an upper diameter which is slightly smaller than the diameter of the adjacent distal end of the sleeve, so that a shoulder is formed at the distal end of the sleeve 21. This shoulder is abutted by a distal end of a pipette tip 20 which is inserted into the pipette tip extension 1 and thus functions as an end stop 23 for controlling an insertion depth of a pipette tip 20. A pipette tip 20 which abuts the shoulder is thereby prevented to further move towards the distal end 3 of the pipette tip extension 1. The sleeve in this case is liquid-tight connected to the end stop whereby a cup like unit is formed having a cylindrical continuation of the connection channel towards the gap. To avoid fluid shortcutting between the pipette tip and the fluid uptake area safely the inner diameter of the sleeve just before the end stop is made to encompasses the pipette tips 20 closely, e.g., having the same or even a slightly undersized diameter with respect to the outer pipette tip diameter.
[0185] FIGS. 14A to 16C show inventive embodiments I9 (FIG. 14A to 14C), I10 (FIG. 15A to 15C), I11 (FIG. 16A to 16C) which surprisingly all worked well with the respective geometries when produced as a one-part pipette tip extension from a respective heterophasic polypropylene copolymer (h-PP) in a hardness range as defined above. This means that the sealing section 19 could despite of the relatively high hardness of the material provide a perfect sealing function during any dispense and aspiration operation when the pipette tip extension of the respective type was slightly pressed towards a clean glass slide or a slide with a test sample 27 applied.
[0186] With all these embodiments however, as shown in FIGS. 14C, 15C and 16C, the lower parting 30 line has been shifted from the usually lowest possible place, here the orifice of the connection channel 16 into the gap 17, whereby surprisingly a more evenly flow of the test liquid in the gap and a correspondingly more uniform washing of the sample from the slide surface could be achieved. To reach this result the parting line should be recessed at least for 0.3 millimeter, preferably for 0.5 to 2.5 mm from the lower orifice of the connection channel. The parting line hereby is an unavoidable line of small imperfections on the surface of an injection molded part, here the pipette tip extension, and forms in the immediate vicinity of a circumferential line where two parts of a mold are separated after the injection molding process. In this case the lower parting line 30 is formed at the inner diameter of a further little step below the end stop 23 of the pipette tip in the connection channel 16. The upper parting line is not shown as being uncritical to the function, e.g. when provided at the exterior wall near or at the proximal end.
[0187] Additionally, and again shown in FIGS. 14C, 15C and 16B, the respective pipette tip extensions can be provided with an optional (as shown in FIG. 15C) cup like unit comprising the sleeve 21 and the guiding plate 14, e.g., similar to that described with FIG. 13B having a cylindrical continuation of the connection channel towards the gap. This adds to a further safety buffer against eventually fluid short cuts 32, despite of the fact, that nearly no short cuts could be observed, even when geometries like shown in FIG. 8 C or 10B have been used with pipette tip extension made from the respective heterophasic polypropylene copolymer (h-PP).
[0188] With FIG. 14C and FIG. 16B a cut out section 29 is shown which extends the height (h) of the gap immediately adjacent to the inner side 6 of the exterior wall 4. In case of FIG. 14C the support bars 15 connecting the guiding plate to the inner side 6 of the exterior wall 4 are provided with a cut out section 29, in case of FIG. 16B the connection elements 28 connecting the inner wall to the sleeve are provided with the cut out section 29 in addition to groove 22 to optimize fluid flow during dispense and aspiration operations.
[0189] For further details which can be combined with the specific features as shown in FIG. 14A to 14C it should be referred to FIGS. 1A to 2B.
[0190] For further details which can be combined with the specific features as shown in FIG. 15A to 15C it should be referred to FIGS. 3A to 4B.
[0191] For further details which can be combined with the specific features as shown in FIG. 16A to 16C it should be referred to FIGS. 4A to 7.
TABLE-US-00001 TABLE 1 Hardness Hardness Hardness TM MFR Density Geometry sealing Ex. Material Tradename Shore A Shore D Rock. R ° C. g/10 min kg/m.sup.3 FIG. +/− Results C1 Body: Borealis Out of >90 112 >121 20 905 8A-C Not formstable also PP1 HF955MO Range relevant under pressure (OoR) C1 Seal: NA elastic elastic elastic NA NA NA 8B, 8C ++ slightly sticky but less relevant as not within the gap C2 PP1 Borealis >>100 >90 112 >121 20 905 9A-C −− No reliable seal; HF955MO C3 TPE1 Mediprene 60 about 16 out of >125 9 890 10A, 10B ++ TPE not hard/ OF601M range stable enough, gap geometry dependent on force; material slightly sticky C4 TPE2 Mediprene 87 about 35 out of >125 12 890 10A, 108 + seal still reliable; OF901M range less deformation; slightly sticky; interfer. with DNA/RNA? C5 TPE2 Mediprene 87 about 35 out of >125 12 890 8A + seal still reliable; OF901M range less deformation; slightly sticky; interfer. with DNA/RNA? C6 PP1 Borealis >>100 >90 112 >121 20 905 10A, 10C − No consistently HF955MO reliable seal C7 PP1 Borealis >>100 >90 112 >121 20 905 11A, 11B − No consistently HF955MO reliable seal C8 LSR Silpuran 70 about 22 out of NA NA 1150 12A, 12B ++ reliable seal but 6000/70 range sticky surface; two components, expens. material I9 PP2 Borealis about 96.3 51 out of 145 1.5 900 ± 10 14A-16C ++ reliable seal; easy SB815MO range to handle; favorable raw material I10 PP3 Borealis about 98.0 53 about 50 141 3.9 900 ± 10 15A-15C ++ reliable seal; easy SC820CF to handle; favorable raw material I11 PP3 Borealis about 98.0 53 about 50 141 3.9 900 ± 10 16A-16C ++ reliable seal; easy SC820CF to handle; favorable raw material Abbreviations Table 1: Ex. Example LSR Liquid Silicone Rubber MFR Melt flow ratio [230° C., 2.16 kg] PP1 Homopolymeric polypropylen 1 Rock. R Rockwell R according to ISO 2039-2 TM Melting temperature TPE1, 2 Thermoplastic elastomer 1, 2 PP2, 3 h-PP 2, 3
TABLE-US-00002 REFERENCE SIGNS LIST 1 pipette tip extension 2 proximal end 3 distal end 4 exterior wall 5 outer side of 4 6 inner side of 4 7 reception aperture 8 dispense aperture 9 inner cavity 10 distance element 11 fluid uptake area 12 constriction element 13 medial axis of 1 14 guiding plate 15 support bars 16 connection channel 17 gap 18 surface of 10 19 seal section 20 pipette tip 21 sleeve 22 groove 23 end stop 24 distance bar 25 inner wall 26 microscopic slide 27 sample 28 connection element 29 cut out section 30 parting line 31 collar (seal) 32 short cut 33 outer stop ring 34 circumferential step 35 shoulder h gap height
