Patent Application
20230302442
The current invention relates to a pipette tip extension according to claim 1 for interacting with areas of interest of biological samples. The pipette tip extension is attachable to a pipette tip.
Different types of pipette tip extensions have been disclosed in detail in WO 2020/131072 A1 and WO 2020/132394 A1 by the same applicant as an alternative device and for an alternative method to specifically address and treat a locally restricted area of interest of a sample, in particular a biological sample, e.g., for histopathological diagnosis or DNA/RNA analysis. As these applications have laid the basis for the present invention, parts of WO2020/132394 have been introduced to the present application for ease of understanding and complementation as will be explained with the brief description of the figures below in detail.
Prototypes of respective pipette tip extensions have been produced in a two-part design where the sealing function of the pipette tip extension towards a sample test slide has been transferred to a separate sealing or gasket, which is a good mechanical solution but ads to the costs if such pipette tip extensions should be used for high volume analytics as with PCR tests or similar.
Many tests with different plastic materials have been made to produce pipette tip extensions in various one-part designs. However, it has been found that form stability and structural robustness, necessary to maintain the flow geometry during dispense and aspiration operations with the pipette tip extension pressed on the test slide, do not go easy with the need to seal the gap between the slide and the pipette tip extension reliably for many plastics. In addition, it was found that some plastics seem to interfere with highly sensible DNA/RNA probes which were dissolved and/or washed from the surface from the test slides with a respective analytical fluid by the dispense and aspiration operation(s). At the same time, it has been found that stickiness of many plastics having good sealing properties is a further obstacle for high precession analytics following the sample extraction due to the risk of dust or particle adhesion on the surface of respective pipette tip extensions.
Further on it has been found that uniform washing and/or dissolving of samples from the surface of the test slides could be detrimental influenced by only minor flow irregularities of the test fluid in the gap between the slide and the pipette tip extension.
It is an object of the present invention to improve the function of known pipette tip extensions and to avoid at least some of the drawbacks as mentioned above, to provide a pipette tip extension which can be easily handled and economically produced.
A pipette tip extension according to the invention is attachable to a pipette tip and comprises a proximal end, a distal end, and an exterior wall extending between the proximal end and the distal end. The exterior wall has an outer side and an inner side and forms at the proximal end a reception aperture for inserting a pipette tip. The exterior wall forms at the distal end a dispense aperture. The pipette tip extension comprises an inner cavity enclosed by the inner side of the exterior wall, and a distance element connected to the inner side of the exterior wall. The distance element is configured to position a pipette tip within the inner cavity and to establish a fluid uptake area adjacent to the inner side of the exterior wall. The fluid uptake area extends from the dispense aperture towards the reception aperture and is in fluid connection with the surrounding atmosphere at the reception aperture. The pipette tip extension is configured as an integrally produced workpiece and made of a non-polar olefine based polypropylene composition. Non-polar herby is used in the usual sense, i.e. the polymer does not show a positive or negative charging. The polypropylene (PP) composition comprises at least 85 wt-%, e.g. from 90-100 wt-%, especially from 95-100 wt-%, 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 polyethylene (PE) dispersed in the matrix, whereat the polypropylene composition has:
In an embodiment of the invention, which may be combined with any other of the embodiments the heterophasic polypropylene copolymer (h-PP) comprises at least 65 to 90 wt-%, e.g. 70 to 80 wt %, especially 76±2 wt-% of the propylene random copolymer matrix phase (r-PPM), whereat the comonomer of r-PPM is ethylene and/or C4 to C12 olefins, e.g. α-olefins, and the h-PP comprises 15 to 35 wt-%, e.g. 20 to 30 wt-%, especially 24±2 wt-% of a dispersed phase comprising an elastomeric propylene copolymer (e-PP) having ethylene and/or C4 to C12 olefins, e.g. α-olefins, as comonomer. By adjusting the content of the e-PP in the matrix, hardness, elasticity, respectively the flexural modulus and the surface friction of the pipette tip extension can be adjusted.
According to the present invention the xylene cold soluble (XCS) fraction of heterophasic propylene copolymer (h-PP), determined at 25° C. according ISO 16152, is regarded as the elastomeric propylene copolymer (e-PP). It is in particular appreciated that the xylene cold soluble (XCS) fraction of the r-PP has a rather low intrinsic viscosity (IV) measured according to ISO 1628-1 (at 135° C. in decalin) from 0.6 to 2.1 dl/g, or 0.8 to 1.3 dl/g which reflects a low average molecular weight.
Further it is preferred that the heterophasic propylene copolymer (h-PP) comprises as polymer components only the random propylene copolymer (r-PP) and the elastomeric propylene copolymer (e-PP). In other words, the heterophasic propylene copolymer (h-PP) may contain further additives but no other polymer in an amount exceeding 5 wt-%, more preferably exceeding 3 wt-%, like exceeding 1 wt-%, based on the total heterophasic propylene copolymer (h-PP), more preferably based on the polymers present in the propylene copolymer (h-PP).
In an embodiment of the invention, which may be combined with any other of the embodiments the material of the pipette tip extension comprises an h-PP whereat the r-PPM has ethylene as comonomer and/or the e-PP has ethylene as comonomer. Therewith the ethylene comonomer content of the r-PPM can be 3 to 20 wt-%, e.g. 5 to 15 wt-%, especially 10±1 wt-% with reference to the r-PPM, and the ethylene comonomer content of the e-PP can be 20 to 40 wt-%, e.g. 25 to 35 wt-%, especially 30±2 wt-%, with reference to the e-PP.
Optionally polyethylene (PE), preferably a low density polyethylene (LDPE) and/or a linear low density polyethylene (LLDPE), may be comprised in the polypropylene composition. The PE again is dispersed in the matrix (r-PPM). Also by adding PE to the polypropylene composition the hardness of the composition can be lowered to a desired value as mentioned above. The content of the PE dispersed in the matrix phase can be equal or less than 15 or 14.9 wt-% of the composition, e.g. 0 to 10 wt-%, especially 0 to 4.9 wt-%, which translates to a weight ration between the heterophasic propylene copolymer (h-PP) and the polyethylene (PE) within the polypropylene composition is in the range of 3/1 to 8/1.
Preferably the polyethylene (PE) has a density measured according to ISO 1180-186 in the range of 903 to 923 kg/m3, more preferably in the range of 910 to 915 kg/m3.
The heterophasic propylene copolymer (h-PP) can be produced in a sequential polymerization process comprising as an example two reactors, preferably connected in series. Accordingly, in the first reactor the matrix (r-PPM) of the heterophasic polypropylene copolymer (h-PP) can be produced and in the second reactor the elastomeric propylene copolymer (e-PP) is obtained. Whereat the r-PPM fraction can be transferred to the second reactor and in the presence of that fracture at least one ethylene and/or C4 to C12 olefin is polymerized to obtain the e-PP.
In an embodiment of the invention, which may be combined with any other of the embodiments the PP composition of the pipette tip extension may further comprise a polymeric α-nucleating agent in a range of 0.0001 to 5 wt-% of the composition. The polymeric α-nucleating agent can be as an example vinyl-alkane- or vinyl-cyclo-alkane polymer.
In a preferred embodiment the PP composition does not comprise any aromatic copolymers or other aromatic compounds contained in the composition.
In a preferred embodiment of the invention, which may be combined with any other of the embodiments, the pipette tip extension further comprises a constriction element which is connected to the inner side of the exterior wall of the pipette tip extension. The constriction element is configured to control an insertion depth of a pipette tip within the pipette tip extension and thereby to define a gap with a gap height between a distal end of an inserted pipette tip and the distal end of the pipette tip extension. The constriction element defines an end stop for a pipette tip and can be configured as a fluid-permeable sieve-like structure.
The proximal end refers in connection with the pipette tip extension to the end of the pipette tip extension which is closest to a reception aperture into which a pipette tip may be inserted. Occasionally, the proximal end might also be addressed to as the upper end of the pipette tip extension. Accordingly, the distal end refers in connection with the pipette tip extension to the end of the pipette tip extension being more distant to the reception aperture for inserting a pipette tip. Thus, the distal end is opposite to the reception aperture, and might also be addressed to as a lower end or bottom end. In connection with a pipette tip, the term proximal end refers to the upper end of the pipette tip which is for use typically attached to a pipette and the associated pipetting channel, while the term distal end refers to the lower end of the pipette tip with an outlet opening for aspirating or dispensing a liquid.
The term “a pipette tip extension attachable/attached to a pipette tip” describes essentially the same situation as the term “a pipette tip insertable/inserted into the pipette tip extension”. It is described here that the pipette tip extension is configured to be connectable to a pipette tip. This may be realized in that, the diameter of the reception aperture and the dimension of the inner cavity formed by the exterior wall is adapted to the dimensions of a pipette tip which shall be inserted into the pipette tip extension. The dimensions of the reception aperture and the inner cavity allow the uptake of a specific pipette tip, and ensure that a gap is formed between the outer wall of the inner pipette tip and the inner wall of the pipette tip extension, into which gap a fluid or liquid can be moved.
A suitable pipette tip may for example be a disposable pipette tip made of a plastic material, or of a so-called fixed pipette tip, made of a metal.
In the context of the present invention, a disposable pipette tip is a pipette tip which may be taken up and/or ejected for example by a liquid handling device automatically. It may be made of a plastic material, and the connection to the liquid handling device may be realized by slightly deforming the plastic to achieve a friction fit connection between the tip and the device.
In the context of the present invention, a fixed pipette tip may be connected to the liquid handling device mechanically by form fit, for example may be screwed to the device. Mounting and dismounting typically requires a manual interaction. A fixed pipette tip may be made of a metal to ensure a stable shape.
The distance element is configured to position an inserted pipette tip at a defined distance to the inner wall of the pipette tip extension, and thereby allows the formation of a fluid uptake area at the inner side of the exterior wall of the pipette tip extension also when a pipette tip is inserted. A distance element thus prevents that an inserted pipette tip fully abuts the inner side of the exterior wall of the pipette tip extension. The distance element therefore function as spacer which allows the fluid uptake area being further limited by a pipette tip when such a pipette tip is inserted into the pipette tip extension.
One distance element is sufficient to space an inserted pipette tip apart from the inner side of the exterior wall for forming a channel between the outer side of the inserted pipette tip and the inner side of the pipette tip extension. This channel may take up a fluid which is to be moved between the pipette tip and the pipette tip extension. A distance element according to the invention may provide a seat for a pipette tip in which the pipette tip does not contact the inner side of the exterior wall.
The distance element is in addition configured to provide a connection with an inserted pipette tip which allows the pipette tip extension being moved by moving the inserted pipette tip, for example under the control of a liquid handling workstation. The connection may for example be a friction fit, which is a force fit connection between the pipette tip extension and an inserted pipette tip, though a form-fit connection may also be possible.
Neither when using one nor when using more distance elements, a distance element does not extend circumferentially along the inner side of the exterior wall, so that the pipette tip extension or the fluid uptake area, respectively, is never closed towards the reception aperture. The reception aperture at the proximal end of the pipette tip extension is during use at least partially open and thus not completely closed, e.g. by a cover, a seal, the pipette tip it is attached to, or a combination thereof. By remaining open to the surrounding atmosphere, sufficient pressure equalization is ensured when a liquid is dispensed from a pipette tip into the fluid uptake area of an attached pipette tip extension.
The distance elements subdivide the inner cavity into a fluid uptake area and a pipette tip hosting area, which correspond to the later position of an inserted pipette tip. The configuration of the distance element and the number used may define the volume of liquid which may be taken up in the fluid uptake area. The fluid uptake area may for example be restricted by the outer wall of an inserted pipette tip and the inner wall of the pipette tip extension, though when the pipette tip extension is configured for treating larger area of interest, additional structures may be used for further restricting the fluid uptake area, as discussed later. Depending on such additional structures, the fluid uptake area may for example extend up to the reception aperture, or may extend simply towards the reception aperture.
In the context of the present invention, a fluid may be any type of liquid or gas, for example a liquid sample, a reagent, a buffer, etc. The fluid may also be a mixture of different liquids (e.g. an emulsion), a mixture of different gases, a mixture of a liquid and a gas (i.e. an aerosol), or a mixture of a liquid and a solid dispensed in said liquid (i.e. a suspension). Solid particles may for example be abrasive particles such a sand, or may be magnetic beads. The liquid may also be a solvent for molecules of interest.
The pipette tip extension is attached to a pipette tip in such a manner that the distal end of both the pipette tip extension and the pipette tip point in the essentially same direction. The same applies to the proximal ends of the pipette tip extension and the pipette tip.
The constriction element functions as a stopper element which is able to restrict the insertion depth of a pipette tip when the pipette tip extension is attached to said pipette tip. The constriction element shall in particular prevent that the distal end of a pipette tip, which is inserted, is positioned flush with the distal end of the pipette tip extension or even protrudes from of the dispense aperture of the pipette tip extension. By restricting the insertion of a pipette tip to a defined depth within the inner cavity of the pipette tip extension according to the invention, a gap, or a corresponding gap height, is defined which is generated when a pipette tip is inserted until the insertion is stopped at the constriction element.
In addition to the stop function, the constriction element is further configured to allow a fluid passing the constriction element when the fluid shall be moved between an inserted pipette tip and the fluid uptake area of the pipette tip extension. This fluid-permeability is achieved be the sieve-like structure of the constriction element. A sieve-like structure in the context of the present invention is understood as a structure having passages for a fluid, while holding back an inserted pipette tip. The sieve-like structure may allow in particular a fluid passaging between an inserted pipette tip and the gap of the pipette tip extension and may further allow a fluid passaging between the gap and the uptake area of the pipette tip extension. The fluid passage may be provided for example by pores or through holes or openings in a structure being otherwise able to hold back an inserted pipette tip from being further introduced into the pipette tip extension. Such a structure may for example be coarse or fine, and for example be a braid, a grid, or a perforated plate.
In the context of the present invention, the distance elements may function here for positioning an inserted pipette within the inner cavity and in relation to the inner side of the exterior wall, thereby allowing that the fluid uptake area is formed without restricting a fluid passage between an inserted pipette tip and the gap. The distance element may additionally serve to hold the position of an inserted pipette tip with respect to the inner side of the exterior wall to a certain extend by providing a certain lateral stability, without hindering a fluid being moved out of an inserted pipette tip towards the gap. The constriction element then may in addition ensure that an inserted pipette tip is hold at a defined height within the cavity and thereby ensuring that a gap is formed which further serves as a fluid passage. The constriction element may be an additional safeguard to ensure a repeatable gap height. The sieve-like structure may then allow a liquid being moved between an inserted pipette tip, the gap, and the fluid uptake area.
The gap height, which is e.g. generated when a pipette tip is inserted under a controlled manner with the aid of the constriction element, may for example be 0.1 mm to 1 mm.
In the context of the present invention, the gap describes the space between the distal end of the pipette tip extension and the distal end of a pipette tip, which is generated when the pipette tip extension and the pipette tip are assembled. The gap serves as a fluid connection between the inner cavity of the pipette tip and the fluid uptake area of the pipette tip extension and is also a part of the fluid uptake area. Upon placing the distal end of the pipette tip extension onto a surface, such as a surface of a microscope slide with a tissue section, the gap may be restricted towards the distal end of the pipette tip extension by closing the dispense aperture of the extension with said surface. Is the dispense aperture of the pipette tip extension closed, a fluid flow between the inner cavities of the pipette tip and the pipette tip extension is possible. Depending on the inner geometry at the distal end of the pipette tip, the volume of liquid and the flow rate can be influenced. A higher flow rate would in particular maximize the shear stress of the liquid on the tissue, while the flow must be sufficient to ensure that the liquid may contact the complete area on the surface of the addressed tissue covered by the dispense aperture of the pipette tip extension.
The use of a constriction element has the advantage that the insertion depth of a pipette tip within the pipette tip extension is controllable by a physical structure, which may be provided in the pipette tip extension. The constriction element according to the invention defines an end stop end for a pipette tip. The end stop provides the stopper function, in that it is for example abuttable by the distal end of a pipette tip when it is inserted into the pipette tip extension.
The control of the insertion depth may ensure that the pipette tip leaves a gap below its distal end when positioned within the pipette tip extension. The gap allows, when the dispense aperture of the pipette tip extension is closed e.g. by a prepared sample or by a part of the surface the sample is provided on, that the liquid may be transported from the pipette tip over the gap and the sample into the fluid uptake area of the pipette tip extension. The gap functions as a fluid connection between the inner cavity of the pipette tip and the fluid uptake area of the pipette tip extension. In an advantageous embodiment, the distance element(s) is or are arranged in such a manner that a continuous fluid uptake area is generated along the inner side of the pipette tip extension, which means that preferably no isolated channels are generated but that each generated fluid uptake area is in fluid connection with the other fluid uptake areas. This ensures that the liquid which is moved between the pipette tip and the pipette tip extension and over the sample remains homogeneously.
In an embodiment of the invention, which may be combined with any other of the embodiments which were or will be mentioned, unless there is a contradiction, the distance element is a sleeve which is configured to receive and circumferentially enclose a distal end of a pipette tip within the inner cavity, without restricting a fluid passage between an inserted pipette tip and the gap.
In this embodiment, the shape and dimension of the sleeve may be adapted to the shape and dimensions of a distal end of a pipette tip which shall be inserted into the sleeve. For example, the sleeve may taper from a proximal to distal direction when the distal end of a pipette tip does so. A sleeve in the context of the present invention is a tubular, elongated envelope or circumferential wall into which at least a distal end of a pipette tip may be inserted. The inner side of the sleeve is thereby contacted by the outer side of the pipette tip. When a pipette tip is inserted into the sleeve, the pipette tip is aligned within the inner cavity of the pipette tip extension. The position and alignment of the pipette tip within the inner cavity of the pipette tip extension may be influenced by the configuration of the shape and dimension of the sleeve as well as by the position of the sleeve within the inner cavity of the pipette tip extension.
In an embodiment of the invention, which may be combined with any other of the embodiments which were or will be mentioned, unless there is a contradiction, the sleeve is open at the distal end for allowing a liquid being dispensed from an inserted pipette tip into the gap.
The configuration of the sleeve may have an influence onto the configuration of the constriction element. For example, it is possible that the sleeve is configured to have the distal end of the sleeve being flush with the distal end of a pipette tip which is inserted into the sleeve. In this case it may be an advantage to arrange the constriction element in an immediate operative connection to the distal end of the sleeve, so that the stop function of the end stop may exercise for example its function upon the distal end of the pipette tip.
It may also be possible that the sleeve is configured to have the distal end of the sleeve being offset from the distal end of a pipette tip which is inserted into the sleeve, the offset being along the medial axis of the pipette tip extension. If for example the distal end of the pipette tip projects out of the distal end of the sleeve, the constriction element may be spaced apart from the distal end of the sleeve, though for example in functional connection with the distal end of the pipette tip. If for example the distal end of the pipette tip is positioned within the sleeve, the constriction element may for example be configured to protrude inwardly into the sleeve.
In an embodiment of the invention, which may be combined with any other of the embodiments which were or will be mentioned, unless there is a contradiction, the sleeve is connected to the inner side of the exterior wall by one or more distance bars which are arranged at the inner side of the pipette tip extension.
By use of such distance bars, for example the position of the sleeve within the inner cavity may be controlled. By use of bar like elements, it may for example also be ensured that the fluid uptake area is in fluid connection with the surrounding atmosphere at the reception aperture.
In an embodiment of the invention, which may be combined with any other of the embodiments which were or will be mentioned, unless there is a contradiction, the pipette tip extension comprises at least two, preferably at least three additional distance elements, each distance element being configured as a distance bar which extends along a direction from the proximal end towards the distal end of the pipette tip extension, wherein the form of the distance bars and the position of the distance bars are configured such that each distance bar can be abutted by an outer side of a pipette tip when inserted into the pipette tip extension.
In this configuration, the additional distance elements may supplement the function of the sleeve of position a pipette tip. Such additional distance elements may have a configuration in which they protrude into the inner cavity. They may here have a surface which is directed towards a medial axis of the pipette tip extension, which can be abutted by an outer side of a pipette tip when the pipette tip extension is attached to the pipette tip. This configuration may be particularly useful if the pipette tip extension is an elongated body (elongated along the medial axis), with a respective elongated inner cavity. In such an elongated embodiment, the additional distance elements may provide a further stability, as an inserted pipette tip is not only positioned by the sleeve but in addition in a more proximal region by such additional distance bars.
An additional distance element may for example be an inner bar, but also other types of protrusions at the inner side of the exterior wall of the pipette tip extension are possible, such as noses, as well as varying forms such as straight or snake- or wave-like bars. Combinations of protrusions of different types or forms are possible. Irregular forms are particularly suitable for providing an additional mixing effect to the liquid which is introduced into the pipette tip extension. It might be possible that only one additional distance element is provided at the inner side of the exterior wall, as long as it still allows the formation of a fluid uptake area in the inner cavity of the pipette tip extension. By coordinating the depth of each additional distance element it may for example be possible to influence the position of an inserted pipette tip within a pipette tip extension in particular in the proximal region of the pipette tip extension, for example in a centric or acentric manner.
In an embodiment of the invention, which may be combined with any other of the embodiments which were or will be mentioned, unless there is a contradiction, the sleeve is arranged centrally within the inner cavity, extending along a medial axis of the pipette tip extension.
A central position may be desired for stability reasons, or so simplify for example the manufacturing process. A central position may be achieved by using additional distance bars for connecting and positioning the sleeve within the desired position in the inner cavity of the pipette tip extension.
It is alternatively possible for example, if the sleeve shall not be connected to the exterior wall by additional distance elements, to connect the sleeve itself to the inner side of the exterior wall. In this situation, also the distal end of the pipette tip would be positioned acentric, i.e. off-center, within the inner cavity of the pipette tip extension.
In an embodiment of the invention, which may be combined with any other of the embodiments which were or will be mentioned, unless there is a contradiction, the sleeve is adapted to the shape of a distal end of a pipette tip.
The adaptation of the sleeve shape, in particular of the inner sleeve shape, to the shape of the distal end of a pipette tip which shall be inserted may provide an increase in the position precision and the stability of the inserted pipette tip within the pipette tip extension.
In an embodiment of the invention, which may be combined with any other of the embodiments which were or will be mentioned, unless there is a contradiction, the constriction element comprises a guiding plate which defines or is connected to the end stop, the guiding plate being arranged close to the distal end of the pipette tip extension but offset from the distal end of the pipette tip extension towards the proximal end, and delimits together with the inner side of the exterior wall the gap at the distal end of the pipette tip extension.
In this embodiment, the length of the offset (along the medial axis) determines the gap height. The guiding plate may be arranged between the distance element, in particular when it is configured as a sleeve, and the distal end of the pipette tip extension, with an offset to the distal end of the pipette tip extension. The guiding plate may guide a liquid which is dispensed by pipette tip through the gap and into the fluid uptake area. The guiding plate may for this have a planar extension, having a disc-like shape, for example, for guiding a liquid. Thus, the surface size of a guiding plate which faces to the gap may be adapted to the lateral extension of the gap, while leaving a fluid passage for example at the outer rim of the guiding plate which faces the inner side of the exterior wall. However, the guiding plate may rather have a more ring-like structure which is for example either directly abuttable by a distal end of an inserted pipette tip, or which is connected to a respective end stop.
A guiding plate may have a shape selected from a ring-shape, a funnel-shape, a plate or disc-like shape, and a shim-like shape. In a plate-like or disc-like shape, the guiding plate preferably comprises a connection channel for providing a fluid passage from an inserted pipette tip into the gap.
A guiding plate may be desired when for example larger amounts of liquids shall be used, for example in the treatment of larger AOI's, and when the size or lateral extension of the dispense aperture is adapted accordingly. Longer distances of a fluid to move from an inserted pipette tip through the gap and into the fluid uptake area typically reduce the flow velocity. This may in turn reduce the effectiveness of the fluid treatment of the sample. By use of a guiding plate, the gap height may be controlled to allow a continuous fluid flow from the pipette tip through the gap into the fluid uptake area of a desired and sufficiently high flow velocity. The use of a guiding plate may also provide a higher stability for a pipette tip extension which is for example adapted to the treatment of a larger AOI.
The guiding plate defines the end stop for example in the case that the guiding plate itself is the end stop. The guiding plate may be arranged below the distance element, in particular below the sleeve, to be abuttable by the distal end of a pipette tip when the pipette tip is inserted into the pipette tip extension. The guiding plate here comprises advantageously a continuous opening which provides a fluid connection from the pipette tip through the guiding plate into the gap.
The guiding plate is connected to the end stop for example in the case where the end stop is a separate structure. Also in this case, the guiding plate comprises advantageously a continuous opening which provides a fluid connection from the pipette tip through the guiding plate into the gap. The end stop may for example be connected to the guiding plate in a way in which a fluid connection from the pipette tip through the guiding plate into the gap is ensured.
In an embodiment of the invention, which may be combined with any other of the embodiments which were or will be mentioned, unless there is a contradiction, the guiding plate extends perpendicular to the medial axis of the pipette tip extension and is connected to the inner side of the exterior wall by one or more support bars, the support bars providing one or more passages for fluidly connecting the gap (17) with the fluid uptake area.
The guiding plate may not necessarily have a perpendicular extension but may be arranged at a different angle, or the guiding plate may have for example a shape which deviates from a plate-like shape, for example a funnel-like shape or others. By the configuration of the shape and dimension of the guiding plate, the flow velocity of a liquid which is used with the pipette tip extension in the treatment of a sample may be influenced.
A support bar may for example be connected to the inner side of the exterior wall and extend laterally with respect to the medial axis of the pipette tip extension. A support bar may alternatively be a protrusion of the exterior wall which extends towards the medial axis of the pipette tip extension. Support bars which connect a guiding plate to the inner side of the exterior wall allow a continuous fluid passage from the gap into the fluid uptake area. The bars may have a more lath-like configuration with a lateral extension, as long as a fluid connection is allowed.
The use of bars to connect the end stop and/or a guiding plate with the exterior wall rather allows the connection being fluid permeable, so that a continuous fluid flow is allowed from an inserted pipette tip through the gap into the fluid uptake area. The use of two or more, or a multitude of bars for example therefore may establish the sieve-like structure of the constriction element.
The one or more support bars may be arranged at the distal end region of the pipette tip extension, but with an offset to the distal end. The one or more support bars may in this case be arranged on the same level as guiding plate. It may alternatively be possible that the one or more support bars are arranged flush with the distal end of the pipette tip extension, and extend inwardly to the guiding plate, which is arranged with an offset of the distal end of the pipette tip extension. The support bars may be arranged in a regular pattern, for example a star-like pattern within the inner cavity of the pipette tip extension, or may for a sieve-like structure of an irregular pattern. The configuration of the one or more support bars may depend on the desired design of the gap at the distal end of the pipette tip extension, or for example on the design of the guiding plate. The support bars may merge with a guiding plate, of they may be configured to carry a guiding plate.
In an embodiment of the invention, which may be combined with any other of the embodiments which were or will be mentioned, unless there is a contradiction, the guiding plate comprises a connection channel which provides a fluid connection between an inserted pipette tip and the gap.
The connection channel is a continuous through opening within the guiding plate which provides a fluid connection from the pipette tip through the guiding plate into the gap. A liquid which is dispensed from an inserted pipette tip may pass the guiding plate and enter the gap by the connection channel. The connection channel may also provide a sieve-like function in that it allows a fluid passage from an inserted pipette tip into the gap.
The connection channel may be arranged on the medial axis of the pipette tip extension, so that the medial axis extends through the connection channel. In this configuration, the pipette tip extension is configured in an axial symmetry, which may be an advantage for the manufacturing process, e.g. in injection molding. However, the connection channel may alternatively be arranged offset of the medial axis of the pipette tip extension, for example in cases where the distance element is configured to position a pipette tip offset of the medial axis, to allow a continuous fluid flow from an inserted pipette tip into the gap. In a further alternative, connection channel and the distance element are configured in a manner in which the distal end of an inserted pipette tip is positioned aside the connection channel. In this case, an additional fluid connector may be used for bridging the offset and for providing the continuous fluid flow from an inserted pipette tip through the guiding plate into the gap.
In an embodiment of the invention, which may be combined with any other of the embodiments which were or will be mentioned, unless there is a contradiction, the end stop is configured as an elongated, tubular hollow cylinder which is functionally connected to the connection channel.
The end stop may be formed by at least one of the following:
The hollow cylinder may serve not only as a stop element but in addition as a fluid connector between the distal end of an inserted pipette tip and the connection channel in the guiding plate. The length of the cylinder may therefore be adapted to the distance between the distal end of an inserted pipette tip and a guiding plate and/or one or more of support bars (which may correspond to the distance between the distal end of the sleeve). The cylinder may therefore be configured as an elongated cylinder, or more flatly as a washer-like (or shim-like) structure.
In a case where the guiding plate forms the end stop, it may be desired that in particular the region around the connection channel serves as a stop. This may for example be achieved in that the diameter of the connection channel is slightly smaller than the diameter of the distal end of the sleeve and/or the diameter of an inserted pipette tip. In this way, the region around the connection channel forms a shoulder which is abutted by the distal end of a pipette tip when it is inserted into the pipette tip extension, so that the insertion depth is limited by the guiding plate.
In the case where the end stop if formed by one or more support bars which extend from the exterior wall forming a ring-shaped stop which is connected to the connection channel of a guiding plate, the ring-shaped stop is abutted by the distal end of a pipette tip upon insertion. The ring-shaped stop may also be considered as a guiding plate, however, as explained before.
In the case where the end stop is formed by a part of a complementary form-fit connection between a pipette tip extension inserted into the sleeve and the end stop, for example a rib or groove of a rib-groove-connection, it may be desired that the sleeve is directly connected to the connection channel in a guiding plate, for example.
In an embodiment of the invention, which may be combined with any other of the embodiments which were or will be mentioned, unless there is a contradiction, the guiding plate comprises a groove which faces the gap at the distal end of the pipette tip extension.
The groove has been shown to reduce the dead volume of liquid when a liquid is aspirated from the fluid uptake area through the gap into an inserted pipette tip. It is currently though that this effect may base on a capillary effect. Dead volume in the present context is understood as being a volume of liquid which remains within the gap for example at the guiding plate although the liquid should have been aspirated into an inserted pipette tip extension.
In an embodiment of the invention, which may be combined with any other of the embodiments which were or will be mentioned, unless there is a contradiction, the groove extends at least in the proximity of the connection channel.
It may be possible that the groove extends circumferentially around the connection channel. It may further be possible that the groove is positioned closer to the connection channel than to the exterior wall or to the outer rim of the guiding plate, which faces to the inner side of the exterior wall.
In an embodiment of the invention, which may be combined with any other of the embodiments which were or will be mentioned, unless there is a contradiction, the distance element is configured as a sleeve for receiving and circumferentially enclosing a distal end of a pipette tip, the sleeve being open at the distal end for allowing a liquid being dispensed from an inserted pipette tip into the gap, and
wherein the end stop is configured as an elongated, tubular hollow cylinder which provides a passage for fluidly connecting an inserted pipette tip with the gap, wherein the sleeve and the end stop are fluidly connected to provide the fluid connection between the pipette tip and the gap.
In a preferred embodiment of the invention, which may be combined with any other of the embodiments which were or will be mentioned, unless there is a contradiction, the sleeve is liquid-tight connected to the end stop to avoid fluid shortcutting between the pipette tip and the fluid uptake area. Therefore, the sleeve can be produced to encompass respective pipette tips closely, e.g., in a friction or form fit manner. For that, the sleeve and the end stop can be formed as a cup like unit with a central dispense aperture which may have a cylindrical continuation of the connection channel towards the gap.
In an embodiment of the invention, which may be combined with any other of the embodiments which were or will be mentioned, unless there is a contradiction, the constriction element comprises a guiding plate which comprises the end stop, the guiding plate being arranged close to the distal end of the pipette tip extension but offset from the distal end of the pipette tip extension towards the proximal end, and delimits together with the inner side of the exterior wall the gap within the inner cavity of the pipette tip extension, and
wherein the pipette tip extension further comprises an inner wall arranged between the sleeve and the exterior wall, wherein the inner wall extends circumferentially around the sleeve and along a direction from the distal end to the proximal end of the pipette tip extension, thereby restricting the fluid uptake area towards within the inner cavity, the inner wall being connected to the exterior wall by the distance bars, and being connected to the guiding plate at the side which faces towards the inner cavity.
In this embodiment, the inner wall encloses the sleeve, and further defined the limitation of the fluid uptake area opposite to the inner side of the exterior wall. By adapting the distance between the inner side of the exterior wall and the inner wall, the size of the fluid uptake area may be controlled independently from the dimension and configuration of the pipette tip which shall be inserted into the pipette tip extension.
The inner wall may extend parallel to the medial axis, though other orientations may be possible, for example to influence the configuration of the fluid uptake area. The inner wall may be connected to the sleeve by connection elements.
In an embodiment of the invention, which may be combined with any other of the embodiments which were or will be mentioned, unless there is a contradiction, the shape and dimension of the dispense aperture is adapted to the size of a sample to be treated (area of interest).
The pipette tip extension of the present invention is particularly useful when an area of interest of a sample shall be treated which has a relatively large size.
In an embodiment of the invention, which may be combined with any other of the embodiments which were or will be mentioned, unless there is a contradiction, the dispense aperture may have a circular form or a circular cross section, respectively, viewed orthogonally to the medial axis of the pipette tip. The dispense aperture is configured to release and receive volumes of a fluid.
In an embodiment of the invention, which may be combined with any other of the embodiments which were or will be mentioned, unless there is a contradiction, the one or more support bars, e.g. a support bar of an off-set rib type, connecting the guiding plate to the inner side of the exterior wall, or a connection element connecting the inner wall to the sleeve have a cut out section provided to extend the height (h) of the gap immediately adjacent to the inner side of the exterior wall to provide a quick and equal liquid transfer within the gap, and between the gap and the fluid uptake area during dispense and aspiration operations of the pipette.
The pipette tip extension according to any of the preceding embodiments can be produced by injection molding of the respective polypropylene composition. Thereby the parting line of at least two mold parts used to produce the pipette tip extension can be shifted into the dispense aperture or the connection channel, so that no press burr or flashes are formed in the transition area from the dispense aperture or the connection channel and the gap or at the surface of the end stop. Thereby flow disturbances can be avoided in the sample are and a good sealing of the pipette tip with the end stop can be provided.
In an embodiment of the invention, which may be combined with any other of the embodiments which were or will be mentioned, unless there is a contradiction, the constriction element is formed by at least one of the following:
The features of the above-mentioned embodiments of the pipette tip extension can be used in any combination, unless they contradict each other.
Another aspect of the invention concerns an assembly comprising a pipette tip for aspirating and/or dispensing a liquid, and a pipette tip extension attached to the pipette tip. The pipette tip extension comprises a proximal end, a distal end, and an exterior wall extending between the proximal end and the distal end. The exterior wall has an outer side and an inner side and forms at the proximal end a reception aperture for inserting the pipette tip, and forms at the distal end a dispense aperture. The pipette tip extension further comprises an inner cavity enclosed by the inner side of the exterior wall, and a distance element connected to the inner side of the exterior wall, for positioning the pipette tip within the inner cavity and for establishing a fluid uptake area adjacent to the inner side of the exterior wall. The fluid uptake area extends from the dispense aperture towards the reception aperture and is in fluid connection with the surrounding atmosphere at the reception aperture. The pipette tip extension further comprises a constriction element which is connected to the inner side of the exterior wall of the pipette tip extension for controlling an insertion depth of the pipette tip within the pipette tip extension. The constriction element defines a gap with a gap height between a distal end of the inserted pipette tip and the distal end of the pipette tip extension. The constriction element comprises an end stop for the pipette tip and is configured as a fluid-permeable sieve-like structure.
The pipette tip extension may comprise one or more features as described above in the context of the single pipette tip extension. These features and/or different embodiments can be used in any combination unless they contradict each other. The definitions given to the pipette tip extension above apply also to the pipette tip extension being part of the assembly.
Another aspect of the invention concerns an injection molding method to produce a pipette tip extension according to any of the preceding claims, comprising:
Further details of the composition as disclosed in context with the description of the PP composition of the pipette tip extension above can be applied. The process could be applied at room temperature, injection pressures between 10 to 35 MPa have been applied depending on the overall dimensions of the pipette tip extension. As an example, for small pipette tip extensions high pressures from 30-35 MPa have been applied, whereas for large pipette tip extensions lower pressures from 10-15 MPa have been applied.
A pipette tip extension made from a material having a hardness as mentioned above could be surprisingly proved to fulfill best the most decisive material relevant functions of a pipette tip extension also with different geometries as described above. The most important and critical functions to be provided by the pipette tip extension geometry and the respective material it is made of have proved to be the following ones, beginning with the most difficult feature first:
Surprisingly it could be shown that a respective polypropylene composition can be tailored to provide as mentioned mechanical features but also to give respective surface friction, without being sticky, to hold the pipette tip extension on the pipette tip and transfer the extension to or from the sample slide, and at the same time allow an easy automatic discharging of the tip extension from the pipette tip. This could be combined best with pipette tip extension of certain geometries as mentioned above and will be explained in detail at the hand of examples and figures.
It should be mentioned that shortcutting, could be also improved by constructional measures, e.g., by providing a sleeve which is liquid-tight connected to an end stop and can be cup like as mentioned above explained at the hand of the figures in detail later. With a combination of a respective h-PP material and a respective liquid-tight connected sleeve the best results with reference to shortcutting could be achieved. Additional surprisingly improvements with reference to the flow characteristic could be achieved by shifting the parting line into the connection channel and/or to provide a cut out section as mentioned above. These measures could help to dissolve and/or solubilize sample material more uniformly from the surface of the test slide, when applied alone or in combination.
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
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.
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”.
The pipette tip extension 1 shown in
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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
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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.
In the embodiment shown in
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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.
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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.
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.
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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.
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.
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
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
With embodiment C3 a pipette tip extension being configured according to
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
Therefore, further geometries according to comparative example C6 with constriction element 4, as shown in
Comparative example C8 using a geometry as shown in
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
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With all these embodiments however, as shown in
Additionally, and again shown in
With
For further details which can be combined with the specific features as shown in
For further details which can be combined with the specific features as shown in
For further details which can be combined with the specific features as shown in
| Number | Date | Country | Kind |
|---|---|---|---|
| 22164100.4 | Mar 2022 | EP | regional |
