Patent Application
20240027305
The presently disclosed subject matter relates to an automated method of dissecting biological material disposed on a planar substrate, such as a glass slide, and transferring the dissected material to a receptacle for further analysis. The presently disclosed subject matter further relates to an associated apparatus and to a disposable tool for use in the apparatus.
To perform a molecular analysis of a tumor, for the purposes of oncology diagnostics, a certain amount and concentration of tumor cells must or should be present in the sample to be analyzed. Tumor tissue is heterogenous and contains other tissue and cell types. Therefore, a region of interest (ROI) is typically defined as the sample to be dissected from a thin section of tissue disposed on a microscope slide. Manual methods of dissection are most common, in which a lab technician scrapes material from the ROI using e.g. a scalpel blade and transfers the scraped material into a collection tube. This can require a great deal of skill and even when performed by a highly skilled technician, there is no guarantee that only material from the ROI will end up in the collection tube.
An enhanced manual method of tissue removal and collection is disclosed in US 2020038001. The method employs suction to remove tissue that has been manually scraped off a slide. The employed dissection tool has three main elements: a shaft part, one end of which can be coupled to a suction source; a cap incorporating a blade at one end and a filter column that is releasably secured to the shaft part via the cap. The filter column includes a filter member which catches dissected material that is suctioned into the tool through the cap. After dissection, the filter column is removed from the tool and e.g. placed in a microcentrifuge tube where the filter member is submerged in a solution for dissolving the dissected tissue.
This method has the advantage of reducing the risk of contamination of tissue samples during the transfer step, but the need for a tool that can be dismantled makes the disclosed method and tool unsuitable for use in an automated device.
An example of an automated device for extracting material from a biological sample via milling is disclosed in U.S. Ser. No. 10/876,933. The device includes a head assembly and a stage that can accommodate and collect samples from several tissue slides and enables automated filling and uploading of several milling tips into a plurality of corresponding sample collection vials. During dissection, the head assembly causes the milling tip to rotate while withdrawing a plunger which simultaneously dispenses buffer solution to the blade part of the milling tool, dissects the tissue and aspirates the buffer solution and dissected tissue into a designated collection vial by depressing the plunger. The used milling tip can then be reloaded into a holder or discarded to avoid cross-contamination.
It is also known to use laser capture microdissection (LCM) to isolate a concentrated population of individual cells or precise anatomical regions of tissue from tissue sections on a microscope slide. A method and device for transferring a microscopic, isolated sample, particularly a membrane-supported micro-dissected specimen, from an object table to an analysis arrangement, is disclosed in U.S. Pat. No. 8,573,073. The device is equipped with nano-suction means and includes a suction tube with a terminal membrane and a vacuum/overpressure unit coupled to the suction tube for sucking or blowing the sample onto or from the terminal membrane. The device and method of U.S. Pat. No. 8,573,073 have the disadvantage that two different tools can be required for dissection and transfer.
There is still room for improvement in terms of defining a more straightforward method, tool and apparatus for dissection and transfer of biological material with minimal contamination that allows the material to be both collected and transferred for analysis in an automated manner, using the same tool.
In a first aspect, the presently disclosed subject matter relates to an automated method of dissecting and transferring biological material from a sample disposed on a planar substrate, such as a glass slide, into a collection tube, using a dissection tool that has an internal passageway extending between first and second ends of the tool and an air-permeable filter element that spans the internal passageway. The tool first end is provided with an orifice and a scraping blade arranged at the orifice. The method includes steps of:
The method of the presently disclosed subject matter enables a “dry” method of transferring the biological material from the glass slide straight to the collection tube, thereby minimizing the risk of contamination. Furthermore, a single tool can be used to dissect, collect and transfer the biological material, which not only simplifies the construction of the equipment that is needed to implement the method in an automated manner, but also further reduces the risk of contamination. The method does not utilize any liquids which would dilute the biological material and impact the methods and quality of the molecular analysis of the collected biological material.
The step of detachment includes bringing the scraping blade into contact with the planar substrate and moving the dissection tool relative thereto. For simplicity, the planar substrate will be referred to hereafter as a glass slide, although as will be understood, any suitable type of substrate on which biological samples are disposed may be used in the method of the presently disclosed subject matter.
Advantageously, the method includes controlling the relative positions of the tool and the glass slide such that the scraping blade engages with the biological material in a region of interest. This optimises the amount and purity of the material that is gathered for analysis.
The step of generating an underpressure suitably includes connecting the internal passageway of the dissection tool, at the tool second end, to a vacuum generator and the step of generating an overpressure includes connecting the internal passageway to a pressure reservoir.
Preferably, the internal passageway of the dissection tool is selectively connectable to the vacuum generator or the pressure reservoir via a valve and the method includes switching the valve from a first position in which the internal passageway is in fluid communication with the vacuum generator to a second position in which the internal passageway is in fluid communication with the pressure reservoir.
The pressure differential between the pressure reservoir and the sealed collection tube creates the pressure pulse that ejects the biological material from the filter element into the collection tube.
In a preferred embodiment, the method includes a further step of using the vacuum generator to at least partially evacuate the collection tube of air before the tool second end is connected to the pressure reservoir. The advantage of evacuating the collection tube is that this increases the pressure differential between the sealed collection tube and the pressure reservoir. A further advantage of evacuating the collection tube is that the atmosphere may serve as the pressure reservoir and the step of generating an overpressure may simply include exposing the evacuated tube to atmospheric pressure.
Preferably, the pressure reservoir supplies a pressure higher than atmospheric pressure, e.g. 200-400 kPa, to ensure that the generated pressure pulse is sufficient to eject the biological material, which may be sticky in nature and adhere to the underside of the filter element. The sealing contact between the collection tube and the connection interface on the dissection tool involves a certain clamping force. Preferably, the pressure reservoir supplies a pressure that is higher than the pressure in the sealed collection tube and lower than the clamping force between the collection tube and the tool.
In embodiments of the method where the step of generating an overpressure involves generating a pressure of greater than atmospheric pressure, the method suitably includes a subsequent step of allowing the collection tube to equilibrate to atmospheric pressure. This can be achieved via controlled leakage of the pressure reservoir, or by switching the valve to a third position in which the internal passageway is in fluid communication with the atmosphere or other suitable method. The collection tube can then be removed from the dissection tool.
Suitably, the method includes controlling the airflow during the step of dissection. In a further development, the method includes a step of measuring pressure or measuring airflow downstream from the filter element of the dissection tool or a step of estimating an area of biological material that has been dissected from the glass slide. The airflow is controlled by adjusting the restriction of a variable restrictor in response to the measured pressure or measured airflow or estimated area of dissected material. The advantage of the further development will be described later with reference to the apparatus of the presently disclosed subject matter.
In a second aspect, the presently disclosed subject matter defines a dissection tool for use in the method, which is disposable and can be manufactured in a straightforward and inexpensive manner. The tool has first and second ends, an internal passageway extending through a body of the tool and an air-permeable filter element arranged in the internal passageway. The tool first end is provided with an orifice and a scraping blade. The scraping blade is preferably made of a metal having a high yield stress. The tool body is preferably made of polymer material. The second end of the tool is provided with a connection interface for releasably connecting the dissection tool in an airtight manner to a mating interface on a tool carrier.
According to the presently disclosed subject matter, the tool body includes a monolithic main body part, i.e. formed as a single piece. The filter element is fixedly retained within the main body part and the mechanical interface includes one or more conical recesses or one or more conical protrusions formed in the main body part.
The tool is intended to be disposed of after use. It is therefore beneficial for the tool to be as inexpensive as possible, which is at least partly achieved via a straightforward manufacturing process. The method of the presently disclosed subject matter enables the dissected material to be removed from the interior of the tool without any need for dismantling of the tool The body part which retains the filter element may thus incorporate the additional functionality of the mechanical interface and may be executed as a single piece, thereby minimising the number of different parts that need to be assembled in order to manufacture the tool.
In one example, the main tool body part also incorporates the scraping blade and the entire tool body is a monolithic part. In a preferred embodiment, which allows enhanced manufacturing accuracy, the scraping blade is provided in a second, “blade” part, which is joined to the main body part. Suitably, the blade part is irreversibly coupled to the main body part, i.e. the two parts cannot be disconnected without breaking the coupling. This may be achieved via a form fit, adhesive bonding or other suitable joining method. The irreversible coupling facilitates robustness of the tool body as a whole. A dissection tool according to the presently disclosed subject matter may thus include a maximum of two body parts.
Preferably, the main tool body part and the blade part, when present, are formed in a moulding process from a suitable polymer material. The metal scraping blade may be embedded, at one side of the tool orifice, or may be attached via adhesive bonding or other suitable joining method. In other examples, the blade part or the entire tool body is overmoulded to the scraping blade. The moulded material can be an injection moulding grade thermoplastic polymer. The grade can be a glass- ceramic-, or carbon-filled material. The polymer can be from the class of polypropylene, polyester, polycarbonate, ABS, or other engineering thermoplastic material and blends of those that are commonly used in engineering and high precision applications
The connection interface for attaching the tool to the tool carrier on a dissection apparatus is integrally formed in the monolithic main tool body part. In order to perform high-precision dissection, it may be important for the tool to be attached with precise alignment, which is achieved via a conical interconnection. In some examples, an upper end of the main tool body part includes a number of conical pins that engage in corresponding conical recesses provided in the tool carrier. Alternatively, the tool may include a number of conical recesses which are adapted to engage with corresponding conical pins on the tool carrier.
In a preferred embodiment, the tool main body part includes a single conical recess which forms part of the internal passageway.
In addition, the second end of the tool may include a click fitting or snap fit elements for establishing a quick coupling with the tool carrier. The aforementioned quick coupling elements are preferably also formed in the main body part of the dissection tool. The advantage of a quick coupling is that automated attachment and detachment of the tool, after dissection, and transfer are facilitated.
The tool further includes a second connection interface, for establishing a seal with the collection tube when it is arranged around the tool orifice. In one example, the second connection interface includes a collar with an outer diameter that is dimensioned to mate with the internal diameter of a collection tube. The collar may include a larger-diameter flange part, an underside of which is used contacting an upper rim of the collection tube when it is placed around the orifice. The seal may then be formed between the upper rim and the aforementioned underside, whereby the collection tube is pressed against the flange part with a certain clamping force to achieve an airtight connection. Suitably, the underside of the flange part may be provided with a layer of relatively soft material to enhance the seal. The collar may also have a conical outer surface that engages with the inner rim of the collection tube in an airtight manner.
Advantageously, for ease of manufacture, the second connection interface is integrally formed with the tool body. It may also be formed by a separate collar made of elastomeric material that is mounted around the tool body. Examples of suitable materials include rubbers (cross-linked, olefins, urethanes or silicones) and thermoplastic elastomers, such as polyesters, polyurethanes, polyolefins. A portion of the collar may have outer diameter that is slightly larger than the internal diameter of a standard-sized collection tube.
The tool of the presently disclosed subject matter further includes an air-permeable filter element arranged within the internal passageway and spanning the full diameter thereof. Suitably, the filter element is made of a porous material with a pore size that does not allow passage of the dissected material, e.g. 10, 50, 100 or 200 m, depending on the sample from which material is dissected. The filter element may have a woven or non-woven structure and is preferably made of a synthetic material such as polyethylene, polypropylene, polyvinylchloride, polyester, nylon, polyvinylidene fluoride or Teflon. The filter element may have a diameter of between 3.0 and 15.0 mm. Alternatively, the filter may be a thin substrate with an array of defined pores made by lithography, etching or laser ablation.
In a preferred embodiment, the filter element material is held in an essentially circular frame part, being a separate part that is inserted into the internal passageway of the main tool body part so as to be fixedly connected therewithin. The frame part may be overmoulded to the filter element material. In one example, the frame part has a larger diameter than the diameter of the tool internal passageway, and is radially retained via an interference fit.
In a further embodiment, the main tool body part is overmoulded to the filter element.
The filter element divides the internal passageway into first and second sections. The first section of the passageway will be defined as the section between the filter element and the tool orifice. The first section of the passageway is preferably straight and has a longitudinal centre axis L. The diameter of the passageway first section may be constant over its length or may vary. For example, the passageway may narrow towards the tool orifice. In use, the part of the tool body that surrounds the first section of the passageway serves as a suction nozzle and will be referred to as the tool nozzle.
The scraping blade is arranged at one side of the tool orifice. The scraping blade has a scraping edge of width w that makes contact with the biological material on the glass slide. In some examples, the blade is a flat, chisel-like part. In other examples, the scraping blade is curved around a blade axis and includes a thin-walled tube section. The tube section may be a continuous thin-walled cylinder or a partial cylinder such as a semi-cylinder. An end face of the tube section serves as the scraping edge.
In some examples of a dissection tool that may be used on an apparatus according to the presently disclosed subject matter, which executes the method of the presently disclosed subject matter, the nozzle is partly formed from a thin-walled tube section and the scraping blade forms part of the nozzle. The tube section may have a wall thickness of 30-100 m and an outer diameter of 3.0-15 mm. The scraping edge of the tube section or the scraping edge of a flat blade can be perpendicular to the blade axis or may be angled for optimized scraping performance. In some examples, the blade has a ground, sharpened edge.
Typically, a dissection tool used in the presently disclosed subject matter is arranged such that the scraping blade engages a horizontal surface of the glass slide at an angle α thereto, which will be defined as a scraping angle. In some embodiments, the scraping blade extends in a direction parallel to the longitudinal axis L of the nozzle and the scraping angle is defined by the orientation of the nozzle axis relative to the slide surface.
In other embodiments, particularly where the scraping blade extends from an end face of the nozzle, the scraping blade has a blade axis that extends at an angle relative to the longitudinal axis L. The nozzle may thus be arranged such that the longitudinal axis L is perpendicular to the slide surface, and the scraping angle is defined by the angle between the slide surface and the blade axis. As will be understood, the scraping angle α can be varied by adjusting the angular orientation of the nozzle axis.
In a further development, the scraping blade of the dissection tool is provided with an anti-stick coating such as NiF or perfluoralkane, to prevent sticking of the dissected material and to ensure that it is suctioned to the underside of the filter element.
In a still further development, the nozzle includes a nozzle extension that at least partially surrounds the tool orifice at a side opposite from the scraping blade. In an example, the tool is adapted to be arranged with a predefined scraping angle α relative to the horizontal slide surface. When the blade is in contact with the slide, a peripheral edge of the nozzle extension may be adapted to create a constant gap to the slide surface. In further examples, the peripheral edge of the nozzle extension has a predetermined profile, such that the gap varies along the circumferential direction.
The advantage of the nozzle extension is to optimize the airstream that is generated during suction, at the location of the scraping edge, thereby ensuring that all or most dissected material is suctioned into the body of the tool.
A dissection tool used in the method of the presently disclosed subject matter thus performs three functions: dissection, collection and transfer. Moreover, since the tool is intended to be disposed of after use, the risk of cross-contamination of the biological material is further reduced. In a third aspect, the presently disclosed subject matter relates to an apparatus for dissecting and transferring biological material from a sample disposed on a planar substrate, which is configured to perform embodiments of the method of the presently disclosed subject matter as described above. The apparatus includes:
In one example, the pressure reservoir is simply the atmosphere. In a further example the pressure reservoir is a pump. In a still further example, the pressure reservoir is a vessel containing pressurized air, held at pressure of 100-400 kPa.
Preferably, the apparatus includes an imaging system for obtaining an image of the biological sample. Suitably, the imaging system is configured to identify the boundary between a region of interest containing biological material to be tested and an unwanted area containing material not to be tested. In other words, the imaging system identifies the shape of the region that is to be dissected. In some examples, the biological sample is a stained tissue sample and the imaging system simply recognizes the stained region. In other examples, the imaging system may be programmed with software to process a captured image of the tissue sample and identify the region of interest based on e.g. cell structure. The system may also be configured to identify the shape of the region to be dissected by comparing the captured image with a reference image that has been marked by a pathologist. Suitably, the imaging system is configured to transfer coordinates of the identified boundary to a controller of the positioning system.
The scraping blade of the dissection tool has a scraping edge of width w. As a result of the relative movement between the blade and the planar substrate in a direction of translation, the scraping edge cuts a track through the biological sample having a corresponding width. When the blade is formed by a section of a thin-walled tube, the width of the track depends on the outer diameter of the tube and the applied contact force.
During dissection, the dissection tool may be arranged such that the scraping blade is arranged at a scraping angle of between 30 and 60 degrees, although other angles may be desirable depending on the nature of the biological sample being dissected.
In a further development, the tool carrier is pivotably mounted to the apparatus to enable adjustment of the scraping angle and, if may be necessary, to bring the dissection tool to a vertical position, to facilitate automated coupling/uncoupling of the dissection tool and/or automated attachment of the collection tube around e.g. the sealing collar of the tool.
Suitably, the positioning system includes motorized actuators for relative movement in transverse X and Y directions and in the vertical direction Z. In some embodiments, the positioning system further includes a rotation stage for adjusting a position of the platform (and biological sample) relative to the scraping blade about a rotation axis that is perpendicular to the planar substrate.
During dissection, the dissection tool is moved relative to the platform in a direction of translation T. In a further development, the tool carrier of the apparatus is rotational about an axis normal to the platform, to enable angular adjustment of the scraping edge relative to the direction of translation. This is advantageous as it allows the effective width of the scraping edge i.e. width measured perpendicular to the direction of translation, to be varied. Suitably, the positioning system includes an actuator for controllable adjustment of angular orientation.
The actuators of the positioning system may be coupled to the platform, for moving the biological sample relative to the dissection tool and/or the actuators may be coupled to the tool carrier for moving the dissection tool relative to the platform.
In a further development, the apparatus is equipped with an air ionizer. Contact between the scraping edge and the planar surface on which the biological material is disposed produces friction, which can lead to a build-up of static charge on the biological material. The air ionizer conditions the air flowing around the dissection tool, and thus reduces any build-up of static charge that could adversely affect the collection of biological material during dissection.
In a still further development, the apparatus is equipped with a variable restrictor arranged in line between the vacuum generator and the filter element of the dissection tool, for regulating airflow through the tool nozzle. In use of the tool, the dissected material collected at the underside of the filter element will restrict the airflow through the filter, thereby increasing the restriction of the system as a whole. This will result in a loss of pressure and a reduction in airflow that could adversely affect the suction capability of the tool nozzle. Suitably, the apparatus is adapted to reduce the restriction of the variable restrictor in response to accumulation of dissected material at the underside of the filter, to enable a constant airflow through the internal passageway and the nozzle.
In one example, the apparatus is further equipped with a pressure sensor or flow sensor arranged in line between the vacuum generator and the filter element and the variable restrictor is controlled based on the measured pressure or measured flow. In a further example, the apparatus is configured to estimate an area of biological material that has been dissected by the scraping blade, and to reduce the restriction of the variable restrictor as a function of the dissected area. The imaging system may be configured to estimate the dissected area or it may be calculated based on the width of the scraping blade and the distance of each translation movement of the blade relative to the slide.
It will be appreciated by those skilled in the art that two or more of the above-mentioned embodiments, implementations, and/or aspects of the presently disclosed subject matter may be combined in any way deemed useful.
The presently disclosed subject matter will now be further elucidated with reference to the embodiments described hereinafter. In the drawings,
It should be noted that items which have the same reference numbers in different figures, have the same structural features and the same functions, or are the same signals. Where the function and/or structure of such an item has been explained, there is no necessity for repeated explanation thereof in the detailed description.
Pathology diagnostic investigation of biological material, such as tissue and cells, forms the basis for many treatment decisions, particularly in oncology. For example, genomic-based tests are performed in order to inform therapy selection for individual patients diagnosed with cancer. The biological material/tissue may be obtained from a biopsy and is then, for example, embedded in paraffin and cut into thin slices which are fixed onto glass slides. These thin slices will be referred to as tissue samples. Other methods of obtaining and preparing biological material are known.
The tissue sample has a region of interest ROI containing material that is to be subjected to the diagnostic testing. The ROI can be identified by staining, or a pathologist may provide markings on a reference slide after analysis under a microscope. The ROI can also be identified via processing of a digital image of the sample. When the ROI has been identified, material is removed/dissected from the slide and then transferred to an analysis arrangement. Typically, the material is transferred to a collection tube, after which a sample preparation process steps, like cell lysis, purification and amplification, etc. and further possible processing steps are performed. As will be understood, the reliability and efficiency of the analysis is optimized, by making sure that only or mostly dissected material from the ROI is present, but also by minimizing contamination.
The presently disclosed subject matter defines an automated method of dissecting biological material from a sample disposed on a planar substrate and transferring it to a collection tube in a manner that permits accurate material removal and minimizes contamination of the material to be analyzed. The method makes use of a dissection tool that is selectively connectable to a source of underpressure and a source of overpressure. The dissection tool and apparatus, which implements the method of the presently disclosed subject matter will be described with reference to
The dissection tool has an essentially hollow body that defines an internal passageway 121 extending between first and second ends of the tool body. An air-permeable filter element 125 is accommodated within the tool body and spans the full diameter of the internal passageway. A first section of the internal passageway is defined between the filter element 125 and an orifice 123 of the tool at the tool first end. The portion of the tool body that surrounds the first section of the internal passageway is defined as a nozzle 122, which has a longitudinal centre axis L.
The nozzle axis L may coincide the with the centre axis of the tool as a whole, as is the case in the example of
In the depicted example, the nozzle is formed by a thin-walled tube and a portion of an end face of the tube serves as a scraping blade 124 (refer
During dissection, the tool is arranged such that the nozzle 122 is oriented at angle α relative to a horizontal surface of the platform 110, which also defines a scraping angle of the blade 124. The tool carrier 130 is pivotably mounted to the positioning unit 135, so as to enable adjustment of the angle α.
The apparatus is further equipped with a vacuum pump 150, a pressure reservoir 160 having a pressure of at least 100 kPa, a valve 140, and suitable tubing arranged such that the internal passageway 121 of the dissection tool 120 is selectively connectable to one of the pump 150 and the pressure reservoir 160. In the depicted example, the valve is a 3-way valve with a first port 141 in fluid communication with the mechanical interface on the tool carrier 130 and with passageway 121, a second port 142 in fluid communication with the vacuum pump 150 and a third port 143 in fluid communication with the pressure reservoir 160.
An embodiment of the method of the presently disclosed subject matter will now be described with reference to
Simultaneously with the step of scraping, an underpressure is generated at the second end of the dissection tool, which creates an uplifting airstream at the tool orifice 123, causing the dissected material to be suctioned into the passageway 121, where it is caught at an underside the filter element 125. During this step of suction, the valve 140 is in a first position in which the first and second ports 141, 142 are in fluid communication and the internal passageway 121 is in communication with the vacuum pump 150. The airstream holds the dissected material at the underside of the filter element 125 during the material removal process.
When dissection is complete, the position of the tool may be adjusted to a transfer position, in which the platform and dissection tool are distanced from each other. In a next step of the method, a collection tube 170 is attached at an outer surface of the dissection tool 120, as shown in
A final step of the method includes exposing the filter element 125 to an overpressure at the second end of tool 120. This is achieved by switching the valve 140 to a second position in which the first port 141 is in fluid communication with the third port 143 and with the pressure reservoir 160, as shown in
The pressure reservoir 160 can simply be the atmosphere. Preferably, to generate a pulse of sufficient magnitude to ensure that all or most material is ejected from the filter element 125, a pump or a reservoir of pressurized air held within a cylinder is used to supply a pressure of 200-400 kPa. Preferably, the supplied pressure is lower than a clamping force generated by the airtight connection of the collection tube 170 around the outer surface of the tool body. This prevents the tube from being released by the pressure pulse, which would create a risk of losing the dissected material.
After exposure to a pressure greater than 100 kPa, the pressure in the collection tube is allowed to equilibrate with the atmosphere. The collection tube may then be detached from the dissection tool 120 and is suitably covered by an end cap.
The method of the presently disclosed subject matter therefore enables automated dissection and transfer of biological material from a sample disposed on a glass slide 115 into a collection tube 170, without the need to use any liquids during dissection or to “rinse” the dissected material out of the tool body. This not only simplifies the process, but also minimizes contamination. An example of a dissection tool 220 suitable for use in an apparatus according to the presently disclosed subject matter is shown in
In use of the tool, a portion of the end face of the nozzle 222 that makes contact with the biological material on the slide 115 serves as the scraping blade 224. The scraping edge 224a in the depicted example is thus arcuate in shape and has a certain width that cuts a track through the biological material when the underside of the nozzle is brought into contact with the slide surface and is moved relative to the slide. The dissected material may therefore have a ribbon shape.
As explained, the underpressure generated by the vacuum pump creates an uplifting airstream that draws in the material dissected by the scraping edge 224a. To ensure that a ribbon of dissected material is suctioned into the tool body, it may be important that the lifting effect of the airstream is optimized at the location of the scraping edge. In the depicted example, this is facilitated by an extension 226 of the nozzle 222 that surrounds approximately half of the tool orifice at a circumferential side opposite from the scraping edge 224a. The extension 226 forms a curved hood, whereby a maximum length of the extension 226, measured in longitudinal direction L relative to the scraping edge 224a, occurs at a circumferential location on the nozzle of 180 degrees relative the scaping edge. At either side of this 180° position, the length of the nozzle extension reduces.
In the depicted example, the dissection tool 220 is adapted to be mounted to a tool carrier, such that the scraping blade 224 is arranged at a scraping angle of 45 degrees relative to the horizontal surface of the slide. A blade axis of the scraping blade extends parallel to a centre axis L of the nozzle and tool, meaning that the scraping angle coincides with the angle of the tool axis relative to the surface of the slide.
The scraping edge 224a forms part of a suction nozzle. A peripheral edge of the extension 226 also forms part of the nozzle and thus shortens the gap G between the nozzle and the slide surface, thereby increasing the suction force exerted on the dissected biological material. The extension 226 is designed such that the gap G is substantially constant when the tool is arranged at 45 degrees to the slide surface and the scraping edge is in contact with the slide. In other examples, the nozzle extension is designed such that the gap varies along the circumference.
In addition to improving the lifting effect at the scraping edge 224a, the extension 226 can also serve as a physical barrier to prevent sideways deflection of a dissected ribbon of material and retain it within the nozzle.
At a second end of the tool 220b, a mechanical interface is provided for releasably attaching the tool 220 at a mating interface on the tool carrier. Suitably, the interface permits an airtight connection to ensure optimal suction through the tool body and a high degree of mechanical alignment, to promote accurate positioning of the scraping edge 224a relative to the slide 115. A click-type fitting, clamping or a bayonet attachment is advantageous for the purposes of speed and automation, but other types of interface, such as threaded connection, are also possible.
The dissection tool 220 further includes an external connection interface for receiving the collection tube that is placed over the tool body. In the depicted example, the outside of the tool body is fitted with a collar 227 made of an elastomeric sealing material for establishing an airtight connection with the collection tube. The dimensions of the collar thus depend on the collection tube used. Preferably, the collar has a chamfered edge 227a for guiding the open end of the collection tube over a main portion 227b of the collar, which has an external diameter that will seal against the internal diameter or rim of the collection tube. Suitably, the collar also has a larger-diameter flange portion 227c that will retain a rim of the collection tube, to prevent damage that might otherwise occur when the tube is attached and evacuated.
The tool further includes a second part 327 which incorporates a nozzle part 322. The second part may be formed from polymer material that is overmoulded to the main body part 326. The scraping blade 324 is embedded in the end face of the nozzle part 322. It is also possible for the nozzle part and the main body part 326 to be entirely formed from material that is overmoulded to the metal scraping blade 324.
In the depicted example, the second part 327 not only includes the nozzle 322 and blade 324, but further includes the second mechanical interface or collar for connection of the collection tube. Like the example shown in
The tool 320 in this example is adapted to be arranged such the longitudinal centre axis L of the nozzle is perpendicular to the glass slide 115 during dissection. At the tool first end 320a, an end face of the nozzle 322 surrounds the orifice 323 at the entrance to the internal passageway 321, indicated via dotted lines. The metal scraping blade 324 is embedded in the nozzle, whereby part of the blade protrudes from the end face of the nozzle. The embedded part of the blade is indicated via dotted lines. In the depicted example, the scraping blade if formed from a thin piece of metal that is bent into semi-cylindrical shape around a blade axis B. The scraping blade may also be a straight, flat part that resembles a chisel.
The blade axis B extends at an angle relative to the nozzle axis L, and the scraping angle is defined by the angle between the blade axis B and the slide surface. The scraping angle can, of course, be varied by adjusting the orientation of the tool body. Suitably, the scraping blade 324 extends towards the nozzle axis L, such that the scraping edge 324a of the blade is located below the tool orifice 323.
A further example of a tool according to the presently disclosed subject matter is shown in
The main tool body part 426 further includes a seat 430 for locating the filter element 425 in axial direction. An example of a suitable filter element is shown in cross-sectional view in
The scraping blade 424 of the tool is provided in a second part 427 that is joined to the main body part 426. Suitably, the second part 427 is overmoulded to the scraping blade 424 and may be irreversible coupled to the main body part 426 via a form fit, adhesive bonding or other suitable joining method. The second part 427 also includes the tool orifice 423 and part of the nozzle 422. An outer surface of the nozzle part 422 in the second body part 427 is adapted to be inserted in a collection tube. The tool body further has a relative flat lower surface 428 for receiving the rim of the collection tube and establishing a sealing connection therewith. This sealing surface 428 can be provided on one of or both of the main body part 426 and the second part 427. Alternatively, the sealing surface may be formed by a conical outer surface of the nozzle part 422.
After dissection and tissue transfer, a dissection tool according to the presently disclosed subject matter is disposed of and a new tool is connected to the apparatus, thereby eliminating the risk of cross-contamination.
Examples, embodiments or optional features, whether indicated as non-limiting or not, are not to be understood as limiting the presently disclosed subject matter as claimed. It should be noted that the above-mentioned embodiments illustrate rather than limit the presently disclosed subject matter, and that those skilled in the art will be able to design many alternative embodiments without departing from the scope of the appended claims.
In the claims, any reference signs placed between parentheses shall not be construed as limiting the claim. Use of the verb “include” and its conjugations does not exclude the presence of elements or steps other than those stated in a claim. The article “a” or “an” preceding an element does not exclude the presence of a plurality of such elements. The presently disclosed subject matter may be implemented by hardware including several distinct elements, and by a suitably programmed computer. In the device claim enumerating several means, several of these means may be embodied by one and the same item of hardware. The mere fact that certain measures are recited in mutually different dependent claims does not indicate that a combination of these measures cannot be used to advantage.
| Number | Date | Country | Kind |
|---|---|---|---|
| 20197369.0 | Sep 2020 | EP | regional |
| 21178992.0 | Jun 2021 | EP | regional |
This application is a national phase filing under 35 C.F.R. § 371 of and claims priority to PCT Patent Application No. PCT/EP2021/075625, filed on Sep. 17, 2021, which claims the priority benefit under 35 U.S.C. § 119 of European Patent Application No. 21178992.0, filed on Jun. 11, 2021 and European Patent Application No. 20197369.0 filed on Sep. 22, 2020, the contents of each of which are hereby incorporated in their entireties by reference.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/EP2021/075625 | 9/17/2021 | WO |
