Apparatus and method for particle isolation

Abstract

An apparatus for particle isolation includes a reservoir for containing a liquid containing particles and a channel with an inlet and an outlet. The inlet is configured to be fluidly coupled with a reservoir containing a liquid containing particles. The apparatus also includes a pressure generator configured to generate a pressure gradient in the channel by which the liquid containing the individual particles is drawn from the reservoir against gravity through the inlet of the channel and causes the liquid to be output through the outlet of the channel. Further, the apparatus includes an actuator configured to induce relative movement between the particles and the inlet of the channel to bring the individual particles into the vicinity of the inlet to be drawn through the inlet.

Description

Technical Field

[0001] Embodiments according to the present invention relate to methods and apparatus for particle isolation, such as a micro-particle isolation platform. Background Art

[0002] Although the use of high-throughput screening platforms (hereinafter referred to as HTS platforms) has been widely established and frequently used for active molecule screening in large-scale pharmaceutical research, highly parallel methods have not yet gained any relevant significance in the bedside environment. This is mainly due to the high system cost of HTS platforms and the lack of integration of the results produced into clinical applications. Early drug screening was limited to active ingredient libraries tested at different concentrations on established tumor cell lines. Successful candidates are then studied in greater depth and, if necessary, supplied to the drug development process. Typically, these steps, biological validation, chemical pharmaceutical modification and translational research phases take seven to ten years (Skardal, 2016). In this process, the success rate of active ingredient development is also very low, because the cell lines used are very imperfect models of physiological tumors. Therefore, HTS is increasingly trying to implement experiments on 3D cultures, spheroids and organ models (Eglen, 2015; Esch, 2015). However, so far, the required high-throughput technology for processing a large number of spheroids or organ models in a fully automated manner has been lacking. Only in recent years, innovative technologies and companies have appeared on the market that promise the first solutions (Zhang, 2017). Therefore, the invention discussed herein represents an important contribution with regard to the establishment of such a high-throughput platform for 3D cell culture.

[0003] In the context of the invention discussed herein, a solution exists for depositing controllable numbers of 3D cell culture models, and more precisely spheres or organoids, in reaction chambers (wells of microplates) with high throughput.

[0004] So far, only three automated single sphere methods have been published, including the technology developed at IMTEK (Gutzweiler, 2017). The published methods for single sphere deposition are based on a print head, which is controlled with an axis robot to predetermined coordinates. The document "Sephor Biomedical GmbH, 2019" discloses a pick-and-place process with a cannula, which aspirates the spheres and can deposit them again. The document "Lang, 2010" discloses a microfluidic dual-chamber separator, which uses a pestle to move individual spheres from a loading chamber to an injection chamber, from which the same can be deposited into droplets. The document "Gutzweiler, 2017" shows non-contact dosing of droplets of single spheres, which have been detected in the nozzle by means of optical detection. Vacuum suction separates droplets that do not meet the detection criteria.

[0005] The Regenova Bio 3D printer (Cyfuse Biomedical K.K., Tokyo, Japan) uses a pick-and-place method (Cyfuse Biomedical K.K., 2019). Here, a cannula uses a vacuum to fix individual spheres from a culture vessel at its inlet. These are then moved to a deposition site by a three-axis robot, where the corresponding spheres can be deposited. This method has also been used by Ayan et al. (2021), who also used a glass cannula to generate a vacuum in order to aspirate and selectively place the spheres (Ayan, 2020). It is important to generate only minimal suction forces, as this will not destroy the spheres. However, the maximum force can vary greatly depending on the cell type affected and will have to be adapted to different patient biopsy samples and cancer types accordingly. If the uptake location (where the spheres are aspirated) and the delivery location (where the spheres are delivered) are too far apart, fast transport speeds are required to enable high throughput. However, with regard to evaporation and drying of biological samples, this is not recommended.

[0006] The spheroid seeding device (Department of Orthopaedic Surgery, University of Otago Christchurch, New Zealand) is able to separate individual spheroids and deposit them again in a controlled manner using a three-axis robot (Lang, 2010). For this purpose, two independent fluid chambers are used, which are connected by a channel. Between the two chambers, spheroid isolation occurs in the fluid channel. This transfers the individual spheroids from the loading chamber to the injection chamber, from which the same can be deposited. This method is difficult to implement when the spheroids present differences in size, which may lead to blockages in the chambers and insufficient isolation. In addition, the design of fluid chambers with different pestles for moving the spheroids represents a challenge for implementation as a cost-effective disposable product or as a sterilizable product.

[0007] Scalable single sphere deposition system (lnstitut für Mikrosystemtechnik IMTEK (Department of Microsystems Engineering), University of Freiburg, Germany) using commercially available, contact-free microdosing equipment (Biofluidix GmbH, Germany) in combination with a particularly developed optical single sphere detection (Gutzweiler, 2017). The spheres are detected in the nozzle and only the tubes that contain exactly one sphere are deposited in a locally controlled manner. The system can be selected according to geometric criteria such as the size and shape of the sphere. Droplets that do not meet the set criteria are sucked away by a vacuum system. All components that come into contact with the liquid are sterilizable disposable products. However, this method has shown that high loss rates are generated by vacuum suction and sedimentation problems in the tube leading to the nozzle cause serious clogging problems.

[0008] According to the prior art, there are some efforts to technically realize the quantitative dosing of small batches of liquids and the isolated distribution of particles.

[0009] Patent TW17184258 describes a fluid dispenser in which a fluid is drawn in by capillary forces and can then be discharged through the fluid dispenser (Taiwan Patent No. TW1718425B, 2017). The basic feature is that the suction device is in the same direction as the fluid dispenser and extends beyond the fluid dispenser. The capillary suction device is a tube with a tapered, hydrophilic interior. According to embodiments of the patent, the spectrum of directional distribution of different fluid volumes, for example in the field of microbiology, is expanded.

[0010] Patent EP2577254A1 describes a device for distributing particles in free-flying droplets (European Patent No. EP2577254A1, 2010). Before the droplets are generated by a piezoelectric drive device, a particle suspension is placed in a non-branched one-way channel. Empty droplets are detected in the one-way channel by an observation device and directed to a rejection position by a specific device. For example, this makes it possible to deposit cells in a microplate in an isolated manner. However, the problem with this method is that no solution is given as to how the particle suspension enters the one-way channel. In order to avoid channel blockage and thus minimize the loss rate, a certain degree of isolation is required, particularly for larger and harder particles, such as spheres compared to single cells.

[0011] Patent AU200823251682 proposes a device for obtaining tissue samples (Australian patent number: AU2008232516B2, 2014). This device, by means of suction, transports a fluid containing various tissue particles through a housing. The device contains two filters, a first filter with a large pore size, which sorts out coarse components from the fluid, and a second filter that filters particles of a certain size. Although a solution for automatic filling of a liquid handling platform by suction is given, this method does not provide a solution for isolating particles in a liquid. Due to the different filters, a large part of the tissue sample is lost, which then has to be reprocessed at great expense.

[0012] Cytostatics are a fundamental pillar of modern cancer therapy and are indispensable for palliative care, especially in advanced stages. Despite scientific rationale and highly sophisticated sequential and parallel treatment regimens with different active ingredients, however, treatment providers and patients face major challenges when patients respond differently to the active ingredients or even develop resistance to the cytostatics used. Even though the collective human genome is more similar (>99%) than different (<1%), these small differences can have far-reaching consequences. Whole genome and epigenomic, transcriptomic and proteomic overview technologies are now available that allow detailed characterization and classification of cancers at the individual level, providing new options for truly customized approaches to treatment and patient care (Bode, 2017).

[0013] In recent years, three-dimensional (3D) cell culture in the form of spheres or organoids taken from cell lines or human tissues has gained significance. In cancer research, solid tumors are no longer understood as groups of homogeneous cell mutants, but as the result of microevolutionary processes (Alberts, 2014). One obstacle to the development of novel therapeutic approaches is the challenge of transferring scientific discoveries from laboratory environments to patients, which is mainly due to the fact that many cancer models do not adequately represent the patient's tumors and many drugs that perform well in cancer models ultimately fail in clinical studies (Ghosh, 2011; Stintzing, 2016). Animal cancer models have undoubtedly provided important insights into the basis of cancer, but they are very time-consuming and not necessarily predictive for human organisms because they cannot faithfully reproduce the pathogenic process of patients. For example, the histological complexity and genetic heterogeneity of human cancers are not typically reflected in transgenic mouse models of cancer (Stintzing, 2016; Caponigro, 2011).

[0014] The main problem known so far for sphere handling platforms is the isolation of spheres from the suspension while keeping the loss rate as low as possible, without restricting flexibility and process control. Summary of the invention

[0015] It is therefore desirable to provide a flexible concept with a high degree of process control which achieves a better compromise between improved isolation of single particles from a suspension, improved transport of the single particles to a desired location and reduced loss rates. The object of the present invention is to provide a liquid handling platform which reliably separates microparticles with a low loss rate, automatically, sterile, flexible, with a high degree of process control and high throughput.

[0016] This is achieved by the subject matter of the independent claims of the present application.

[0017] Further embodiments according to the invention are defined by the subject matter of the dependent claims of the present application.

[0018] The approach to personalized medicine discussed in this article aims to minimize the use of animal testing by not focusing on (mouse) 2D cell culture, but using 3D cell culture models (spheroids, organoids) from samples of patients in a more targeted manner. Patient-specific spheroids, such as tumor spheroids, can be used to gain important insights into individual treatments and new drug development in general. This means that animal testing can be used in a more targeted way in (pre) clinical research and drug development in the future, or, if possible, can even be avoided. With the help of an automated processing platform based on a liquid handling system, spheroids are placed individually and in a controlled manner in separate wells of a microplate, and then treated with different active ingredients and concentrations of chemotherapy. Automatic spheroid processing is to compensate for the current shortcomings of previous technologies. The spheroid processing methods used so far for translational clinical research involve a lot of manual labor. This limits the number of possible and technical repetitions in in vitro experiments and leads to limited statistical significance.

[0019] In summary, it can therefore be stated that the invention discussed herein substantially exceeds the prior art presented in the following respects:

[0020] - Miniaturization, parallelization, and automation for high-throughput in vitro experiments on spheroids and organoids.

[0021] - The platform is also suitable for high-throughput screening (HTS) in terms of throughput and automation, unlike other microfluidic technologies.

[0022] - A high degree of flexibility and process control is enabled by guiding specifically selected spheroids / tissue-like objects from the reservoir to the target location.

[0023] According to one aspect of the present invention, the inventor of the present application realizes that a problem encountered when trying to guide a single particle to a target position originates from the fact of particle deposition. According to this aspect of the present application, this difficulty is overcome by suctioning a single particle against gravity. This is based on the idea that such transportation resists the deposition of particles and therefore prevents the blockage of the channel through which the particles are transported. In particular, active suction is advantageous because the applied force overcomes the passive force acting on the particles, and compared to passive transportation (like transportation using capillary forces), it can make particles transported faster. The use of active suction acting on the resistance to gravity causes the effective transportation of microparticles through the channel and can be high-throughput. Further, the inventor finds that it is highly effective to use the same machine or equipment for the inlet of the suction particle through the channel and for the outlet of the liquid through the channel with or without a single particle. The idea based on this is that, for example, the suction pointing to the output direction can be applied to the output of the channel, causing the output of the fluid through the outlet and the suction of the fluid through the inlet. This suction applies enough force on a single particle to be sucked through the entrance against gravity, and the force applied is small enough not to destroy or break the particle. In addition, this can be a small system because it does not need to provide a separation device or machine for suction and output. Another advantage is the use of a channel for the transportation of particles from a reservoir to a target location, because the particles are suspended in a liquid during the entire transportation process. Therefore, the channel prevents evaporation and drying during transportation, and therefore also allows the long distance between the reservoir and the target location and the rapid transportation of the particles sucked. Further, the device uses the relative motion between the particles in the reservoir and the entrance of the channel to be able to suck a single particle through the entrance and, for example, to allow a specific selection of particles to be sucked. Therefore, it is possible to transport a single particle to their respective target locations individually. The present invention is a concept for separating particles (like cells, spheroids or organoids), in a liquid medium in a reservoir by sucking particles individually from a reservoir, passing them through a capillary or capillary, and transporting them to the target location.

[0024] Accordingly, according to this aspect of the present application, a device for particle isolation includes a reservoir for accommodating a liquid containing particles and a channel with an inlet and an outlet. The inlet is configured to be fluidically connected to the reservoir. For example, a channel with a channel portion including an inlet can be immersed in the reservoir. The channel portion including the inlet can be immersed in the liquid containing particles in the reservoir. Alternatively, the channel can be fixed to the reservoir. Further, the device includes a pressure generator, which is configured to generate a pressure gradient in the channel, through which the liquid containing a single particle is sucked from the reservoir against gravity through the inlet of the channel. In addition, the pressure generator is configured to cause the liquid to be output through the outlet of the channel. The pressure generator can be a pump, a piezoelectric driven plunger or a hot droplet generating device. In addition, the device includes an actuator, which is configured to cause relative motion between the particle and the inlet of the channel so as to bring the single particle to the vicinity of the inlet to be sucked through the inlet.

[0025] According to an embodiment, the channel portion including the inlet may be arranged at an angle of at least 10° with respect to the bottom surface of the reservoir. This enables, for example, the suction of particles through the inlet of the channel against gravity. The inventors have found that it is particularly advantageous if the channel is arranged at an angle of 30° or 90° with respect to the bottom surface.

[0026] According to an embodiment, the actuator is configured to move the inlet of the reservoir and / or the channel so as to bring a single particle (preferably a deposited particle) into the vicinity of the inlet to be sucked through the inlet. The single particle, for example, is a particle deposited on the bottom surface of the reservoir. The actuator, for example, is a motorized positioner, such as a three-axis stage or a robot, configured to move the reservoir with a single particle so as to bring a single particle into the vicinity of the inlet to be sucked through the inlet. Alternatively, the actuator, for example, is a motorized positioner, such as a three-axis stage or a robot, configured to move the inlet of the channel so as to bring a single particle into the vicinity of the inlet to be sucked through the inlet. According to a preferred embodiment, the actuator includes a first actuator, the first actuator is configured to move the reservoir in two dimensions, such as a first dimension representing the x direction and a second dimension representing the y direction; and a second actuator, the second actuator is configured to move the channel in a third dimension, such as in the z direction, perpendicular to the plane defined by the two dimensions (i.e., defined by the first dimension and the second dimension), such as the xy plane. For example, the first actuator and the second actuator are configured to bring a single particle specifically / purposefully into the vicinity of the inlet to be sucked through the inlet. The first actuator, for example, moves the reservoir to bring a specific particle to the same (x, y) position as the inlet of the channel, and the second actuator moves the channel to reduce the height difference between the specific particle and the inlet of the channel. The movement of this combination of the reservoir and the channel effectively brings the specific particle into the vicinity close to the inlet to suck the specific particle through the inlet. With the device, the movement of the inlet of the reservoir and / or the channel enables selective suction of particles, particularly deposited particles.

[0027] According to an embodiment, the actuator is configured to induce a mixed liquid flow in the reservoir to prevent particles from being deposited in the reservoir and to bring the particles individually to the vicinity of the entrance of the channel. For example, the actuator can be a pump configured to induce a mixed liquid flow or a vibration device configured to vibrate the reservoir to induce a mixed liquid flow or a magnetic device configured to move magnetic particles in the reservoir to induce a mixed liquid flow. The induced mixed liquid flow moves particles in the liquid in the reservoir. The particles are lifted to the entrance of the channel to be sucked. With such an actuator, particles can be easily and effectively brought to the entrance of the channel. In addition, the mixed liquid flow can prevent particles from aggregating and enable the suction of individual particles that can pass through the entrance of the channel.

[0028] According to an embodiment, the reservoir is a reservoir channel within a microchip, and the inlet is fluidically connected to the end portion of the reservoir channel. The reservoir channel can represent a microchannel, which represents a channel with a diameter of less than 1 mm in microtechnology. An actuator, such as a flow generator, can be configured to cause the flow of liquid in the reservoir channel to bring particles individually to the vicinity of the inlet of the channel. For example, the actuator is configured to cause the flow of particles, and the particles are transported to the inlet of the channel one by one by aligning the reservoir channel. The actuator can be configured to apply a sheath fluid in a laminar flow to concentrate the particles in the center of the fluid or stream. Optionally, the reservoir channel can have a specific shape, such as a tortuous shape, so that the flow caused by the actuator in the reservoir channel enables particle individualization.

[0029] According to an embodiment, the device includes a reservoir monitoring device, such as a camera and / or a microscope and / or an impedance sensor, configured to monitor a reservoir, such as a portion of a reservoir or the entire reservoir, to obtain information about the position of a single particle in the reservoir (e.g., the reservoir position), i.e., the first information. It is also possible that the reservoir monitoring device is configured to obtain further information, such as the position of other particles, the number of particles, the size of the particles, the shape of the particles, etc. The actuator is configured to cause relative motion based on the information about the position of the single particle to bring the single particle to the vicinity of the entrance. For example, the actuator is configured to move the entrance of the channel to the position of the single particle. In this case, the actuator can be a three-axis motorized positioner, such as a stage or a robot, configured to move the portion of the channel including the entrance relative to the single particle in the reservoir. Alternatively, the actuator is configured to move the reservoir relative to the portion of the channel including the entrance so as to position the reservoir so that the entrance of the channel is at the position of the single particle. In this case, the actuator may be a three-axis motorized positioner, like a robot or a stage, configured to move the reservoir with the single particle at a position defined by the information about the position of the single particle. Alternatively, it is also possible that the actuator may include a first actuator configured to move the reservoir and a second actuator configured to move the channel, wherein the actuator uses the information about the position of the single particle to control the first actuator and the second actuator in order to bring the single particle into the vicinity of the inlet. This feature enables selective aspiration of individual particles.

[0030] According to an embodiment, the reservoir monitoring device is further configured to obtain information, such as second information, about the size and / or shape of particles positioned in a monitoring area of ​​the reservoir monitoring device. The device may also include a processor configured to determine a respective particle type for each particle positioned in the monitoring area based on the information about the size and / or shape of the particles positioned in the monitoring area. The processor may be configured to select particles of a predetermined particle type from the particles positioned in the monitoring area as individual particles to be aspirated and provide information about the position of the individual particles to the actuator. In the case where different types of particles are suspended in the liquid in the reservoir, it is therefore possible to classify the particles by aspirating only particles of a predetermined type.

[0031] According to an embodiment, the reservoir monitoring device is configured to obtain information about the position of particles located in the monitoring area of ​​the reservoir monitoring device. The device may also include a processor, such as the processor described above, which is configured to determine the distance between particles located in the monitoring area based on the information about the position of the particles located in the monitoring area. The processor may be configured to select a single particle from the particles located in the monitoring area to be sucked according to the determined distance between the particles, and provide information about the position of the single particle to the actuator. For each neighboring particle of the single particle to be sucked, the respective distance between the single particle to be sucked and the respective neighboring particle, for example, must be equal to or greater than a predetermined threshold, for example, a predetermined distance threshold. For example, the processor is configured to select a single particle to be sucked by checking whether the distance between the first particle and its neighboring particles is equal to or greater than a predetermined threshold, and if all distances between the first particle and other neighboring particles are greater than the predetermined threshold, by selecting the first particle as the single particle to be sucked. For example, the predetermined threshold is pre-set. The predetermined threshold value may depend on the type of particles to be drawn, the size of the cross section of the channel and / or the pressure gradient caused by the pressure generator. The predetermined threshold value may be equal to 50 microns, 100 microns, 250 microns, 300 microns, 500 microns or 1 millimeter. By checking the distance between particles, it can be ensured that only one particle is drawn through the inlet and there are no other adjacent particles.

[0032] According to an embodiment, the device includes a monitoring device, such as a camera and / or a microscope and / or an impedance sensor, configured to monitor the vicinity of the entrance of the channel, such as the entrance, or the portion of the channel including the entrance, and the surrounding environment of the entrance in the reservoir. The monitoring device may correspond to the reservoir monitoring device described above. The monitoring device may be configured to obtain information about whether a single particle is present in the vicinity of the entrance, and / or information about the size and / or shape of a single particle in the vicinity of the entrance, and / or information about whether a single particle is sucked through the entrance, and / or obtain information about the time instant at which a single particle is sucked through the entrance. The device also includes a controller, and if a particle is detected near the entrance of the channel by the monitoring device, the controller is configured to control the pressure generator to cause a pressure gradient in the channel. This monitoring near the entrance of the channel ensures that only one single particle is sucked through the entrance.

[0033] According to an embodiment, the device includes a monitoring device or the monitoring device described above. The monitoring device is configured to obtain information about the size and shape of a single particle (i.e., a single particle to be sucked through the inlet near the inlet), and information about the time instant at which the single particle is sucked through the inlet. In addition, the device includes a calculator configured to determine the position of the single particle (i.e., the inhaled particle) in the channel based on the information obtained by the monitoring device, i.e., the information about the size and shape of the single particle and the information about the time instant at which the single particle is sucked through the inlet, and based on information about the flow velocity through the channel. The flow velocity, for example, is given or controlled by a pressure generator. Alternatively, it is also possible to obtain the flow velocity and determine the flow velocity, and the flow velocity comes from a channel monitoring device configured to monitor a predetermined volume of the channel. This enables the tracking of the single particles in the channel and the controlled transportation of the single particles to their designated locations.

[0034] According to an embodiment, the device includes a decision-based injection device, such as a discarding device, configured to determine information about whether a single particle (i.e., a single particle that is sucked) is present in a predetermined volume of the channel located near an outlet of the channel based on the position of the single particle in the channel obtained from a calculator. The information indicates whether the single particle is near the outlet. The predetermined volume can indicate the volume of liquid to be output in the channel through the outlet with the next injection by the pressure generator. The calculator, for example, is configured to determine the position of the single particle at different time instants to track the transportation of particles through the channel. The decision-based injection device, for example, is configured to determine the time instant at which the single particle is located in the predetermined volume of the channel based on the position of the single particle in the channel obtained from the calculator. The decision-based injection device is configured to inject liquid without single particles to a first target location, and liquid containing single particles to a second target location. The decision of whether the liquid in the predetermined volume is injected to the first target location or to the second target location depends on whether the single particle is in the predetermined volume of liquid. For example, the decision-based injection device is configured to inject liquid to a first target location until the liquid containing a single particle is to be output through the outlet. Then, the decision-based injection device is configured to inject the liquid containing the single particle to a second target location. This allows the single particle to be accurately transported to the target location and reduces the possibility of erroneously injecting (i.e., to the wrong target location) liquid with or without particles.

[0035] According to an embodiment, the device includes a channel volume monitoring device, i.e., a channel monitoring device, configured to monitor a predetermined volume of a channel located near the outlet of the channel and obtain information about whether a single particle (e.g., a single particle sucked) is present in the predetermined volume. The predetermined volume can be defined as described above. If the information about whether a single particle is present in a predetermined volume indicates that a single particle is not present in the liquid to be output, the decision-based injection device is configured to inject a liquid without a single particle to a first target position; if the information about whether a single particle is present in a predetermined volume indicates that a single particle is present in the liquid to be output, the decision-based injection device is configured to inject a liquid containing a single particle to a second target position. The liquid to be output, for example, is a liquid within a predetermined volume of the channel. The channel volume monitoring device reduces the possibility of mistakenly injecting (i.e., to a wrong target position) a liquid with or without particles. In addition to the calculator described above for determining the position of a single particle in the channel, the device may include a channel volume monitoring device. The two systems can work together to further improve the accuracy of the injection of the liquid to its target position.

[0036] According to an embodiment, the device includes a particle concentration monitoring device configured to determine the particle concentration in the liquid containing particles in the reservoir and indicate whether the particle concentration exceeds a predetermined threshold, such as a predetermined concentration threshold. A high concentration of particles increases the probability that more than a single particle is aspirated. Therefore, the particle concentration monitoring device reduces the probability that two or more particles are aspirated instead of only one particle. If the particle concentration monitoring device indicates that the particle concentration exceeds a predetermined threshold, the person using the device can adjust the concentration so that the concentration is below the predetermined threshold. Optionally, the device may also include a concentration adjustment device, which is configured to reduce the particle concentration of the liquid in the reservoir if the particle concentration monitoring device indicates that the particle concentration exceeds a predetermined threshold. For example, the concentration adjustment device may be configured to add liquid, i.e., liquid without particles, to the reservoir to reduce the particle concentration. The predetermined threshold may be equal to 50 particles / ml, 100 particles / ml, 500 particles / ml, 1000 particles / ml, 5000 particles / ml, or 10000 particles / ml. The predetermined threshold, such as a predetermined particle concentration threshold, may depend on the size and / or geometry of the reservoir.

[0037] According to an embodiment, the channel is a continuous filter-free channel. The device is configured to transport specific single particles, for which reason no filter needs to be integrated in the channel. Furthermore, it is advantageous that the channel is a continuous channel, since such a channel is easy to clean / sterilize. Even if some particles will be deposited in the channel, these deposited particles can be easily removed from the continuous filter-free channel, since no interfering structure is integrated in the channel.

[0038] According to an embodiment, the channel is, for example, a disposable hose or tube having a uniform diameter over its entire length. Since the hose or tube can be easily replaced with a new one, the device is easy to implement and can easily meet hygiene standards. Even if some particles are deposited in the channel, these deposited particles can be easily removed from the channel due to the uniform diameter.

[0039] According to an embodiment, the pressure generator is configured to output a single particle passing through an outlet of the channel by distributing the single particle in a free-flying droplet. For example, the pressure generator includes a pressure actuator, which is configured to press against the channel and deform it to eject the liquid containing the single particle in the droplet. The pressure actuator is configured to release the channel after pressing against the channel to generate a pressure gradient, i.e., a pressure gradient in the direction of the outlet. The channel can be other than a deformable material, such as a polymer material.

[0040] Another embodiment is directed to a method for particle isolation, the method comprising the steps of fluidly coupling an inlet of a channel to a reservoir holding a liquid containing particles, and inducing relative motion between the particles and the inlet of the channel so as to bring a single particle into proximity with the inlet to be drawn through the inlet. The method further comprises generating a pressure gradient in the channel through which the liquid containing the single particle is drawn from the reservoir against gravity through the inlet of the channel and outputting the liquid through an outlet of the channel.

[0041] The method as described above is based on the same considerations as the device as described above. In this way, the method can have all the features and functions that are also described in the device, wherein the features and / or functions correspond to the method steps and / or features of the method steps. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] The drawings are not necessarily to scale, emphasis instead generally being placed upon illustrating the principles of the invention. In the following description, various embodiments of the invention will be described with reference to the following drawings, in which:

[0043] Figure 1 An embodiment of an apparatus for particle isolation is shown;

[0044] Figure 2 A more detailed embodiment of an apparatus for particle isolation is shown;

[0045] Figure 3 An actuator configured to move an inlet of a reservoir and / or a channel is shown;

[0046] Figure 4 A microchip connected to an inlet of a channel of the device is shown;

[0047] Figure 5 An actuator is shown, which is configured to induce a mixed liquid flow in a reservoir using a pump;

[0048] Figure 6 An actuator is shown, which is configured to induce mixed liquid flow in a reservoir using magnetic principles;

[0049] Figure 7 An embodiment of a droplet generating device is shown as an example of a pressure generator;

[0050] Figure 8 A pump is shown as an example of a pressure generator;

[0051] Fig. 9 An embodiment including a collection chamber having a functional structure is shown; and

[0052] Fig.10 A block diagram of a method for isolating particles is shown. DETAILED DESCRIPTION

[0053] In the following description, the same or equivalent elements or elements having the same or equivalent functions are denoted by the same or equivalent reference numerals even if they appear in different drawings.

[0054] In the following description, a number of details are presented to provide a more comprehensive explanation of embodiments of the present invention. However, it is apparent to those skilled in the art that embodiments of the present invention can be practiced without these specific details. In other examples, well-known structures and devices are shown in the form of block diagrams rather than in the form of details to avoid obscuring embodiments of the present invention. In addition, unless otherwise specifically stated, the features of the different embodiments described later herein may be combined with each other.

[0055] Figure 1 An embodiment of an apparatus 1 for particle isolation is shown. The apparatus 1 comprises a reservoir 15 for containing a liquid 16 containing particles 17. In the reservoir 15, the particles 17 have the same or higher density than the liquid 16. Furthermore, the apparatus 1 comprises a channel 10 with an inlet 11 and an outlet 12. The channel 10 may be a continuous filter-free channel. The channel may be a hose having a uniform diameter over its entire length.

[0056] The inlet 11 of the channel 10 is configured to be fluidically coupled to the reservoir 15. For example, the portion of the channel 10 including the inlet 11 may be dropped into the reservoir. Figure 1 As shown, the channel 10 can extend into the reservoir 15 so as to fluidically connect the inlet to the reservoir. For example, the support structure can be configured to maintain the portion of the channel 10 including the inlet 11 in a stationary position, wherein the portion of the channel 10 including the inlet 11 extends into the reservoir 15. Alternatively, for example, an actuator, such as a robot or a stage, can be configured to move the portion of the channel 10 including the inlet 11 to fluidically connect the inlet 11 to the reservoir 15. The portion of the channel 10 including the inlet 11 can be arranged in a stationary manner or in a movable manner.

[0057] Furthermore, the device 1 comprises a pressure generator 14, which is configured to generate a pressure gradient in the channel 10, by which a liquid containing a single particle 17a is sucked 13a from a reservoir 15 against the gravity 2 through the inlet 11 of the channel 10. The suction against the gravity 2 has the effect of reducing the probability that a plurality of particles blocking the channel are sucked. The suction against the gravity 2 increases the probability that only one particle is sucked, for example, at each instant in time. The single particle 17a represents one of the particles 17 within the liquid 16 in the reservoir 15. The pressure gradient force caused by the pressure gradient acts on the single particle 17a so as to suck the single particle 17a. The direction of the pressure gradient force is along the channel 10 from the inlet 11 of the channel 10 to the outlet 12 of the channel 10. According to the pressure gradient, a low pressure is achieved in the portion of the channel 10 including the outlet 12, and a high pressure is achieved in the portion of the channel 10 including the inlet 11. The pressure gradient force is directed from an area of ​​high pressure to an area of ​​low pressure, i.e. from the inlet 11 to the outlet 12.

[0058] like Figure 1 As shown, the portion of the channel 10 including the inlet 11 is arranged at a 90° angle with respect to the bottom surface 15a of the reservoir 15. Therefore, the single particle 17a must be sucked 13a against the gravity 2. Another angle is also possible, and the inventors have found that an angle of at least 10°, 20° or 30° achieves a good suction 13a of the single particle 17. This is based on the fact that such an angle enables controlled, active and specific suction of the single particle. Such an angle enables easy implementation of the relative movement caused by the actuator 30, for example, in particular the relative movement between the reservoir 15 and the inlet 11. Moreover, such an angle reduces the possibility of additionally sucking adjacent particles 17 (i.e., particles 17 adjacent to the single particle 17a in the reservoir 15). With regard to the possibility of sucking 13a only one single particle 17a passing through the inlet 11 and with regard to the efficiency of the transport of the single particle 17a through the channel 10, a 90° angle is particularly advantageous. Angle α can be particularly advantageous in the range of 10°≤α≤90°.

[0059] like Figure 1 As shown, a single particle 17a pumped 13a by a pressure generator 14 through an inlet 11 is transported through a channel 10. Hereinafter, a single particle 17a in a channel 10 may be understood as a pumped particle 17b. The pressure gradient in the channel 10 induces a flow 21 that transports the pumped particle 17b to the outlet 12 of the channel 10.

[0060] In addition, the pressure generator 14 is configured to output 13b of liquid 16 with or without individual particles 17a (i.e., aspirated particles 17b) through the outlet 12 of the channel 10. The pressure generator 14 can be configured to output 13b a predetermined volume of liquid 16 through the outlet 12 at a certain moment in time. The pressure generator 14 can be configured to first output 13b a predetermined volume of liquid 16 with or without individual particles 17a through the outlet 12, and then aspirate 13a an equal amount of liquid with or without individual particles 17a through the inlet 11. Therefore, the output 13b and the aspiration 13a of the pressure generator 14 work together to achieve efficient transportation of the individual particles 17a from the reservoir 15 to the target location. The output 13b and the aspiration 13a can be performed by the pressure generator 14 as required.

[0061] As about Figures 7 to 9 The pressure generator 14 may be implemented as described in one of the embodiments.

[0062] Furthermore, the apparatus 1 comprises an actuator 30 configured to induce a relative motion 18 between the particle 17 and the inlet 11 of the channel 10 so as to bring a single particle 17a into the vicinity 32 of the inlet 11 to be pumped 13a through the inlet 11. To achieve the relative motion, the channel 10, the reservoir 15 and / or the particle 17a itself may be moved by the actuator 30. The actuator 30 may comprise one or more

[0063] - a motorized positioner for moving the channel 10 in the z dimension, the xy dimension or the xyz dimension;

[0064] - a motorized positioner for moving the reservoir in the z dimension, the xy dimension, or the xyz dimension;

[0065] - a vibrating device for moving the reservoir; and

[0066] - A flow generating device for inducing a microfluidic flow or a mixed flow in the reservoir 15 .

[0067] Can be Figure 1 Different types of actuators implemented individually or in combination in the device Figures 2 to 6 For example, a flow generating device, a vibration device, or a motorized positioner for moving the reservoir 15 can be combined with a motorized positioner for moving the channel 10.

[0068] The pressure generator 14 may be configured to generate a pressure gradient in the channel 10 as required (e.g., at a time instant when a single particle 17a is in the vicinity 32 of the inlet 11). For example, the apparatus 1 may include a monitoring device configured to monitor the vicinity 32 of the inlet 11 and, at a time instant when a single particle 17a is detected in the vicinity 32 of the inlet 11, trigger the pressure generator 14. The pressure generator 14 may be configured to generate a pressure gradient in response to a trigger received from the monitoring device.

[0069] like Figure 1 As shown, the pressure generator 14 can be arranged at the outlet 12 of the channel 10. Alternatively, it is possible that the pressure generator 14 is arranged near the outlet 12 of the channel 10, for example, the outlet 12 of the channel 10 can be connected to a chamber, the liquid 16 with or without individual particles 17a is output into the chamber through the pressure generator 14, and the pressure generator 14 can be connected to the chamber to generate a pressure gradient in the channel 10 above the chamber, for example, see Figure 8 and Fig. 9 .

[0070] The present invention is a concept for isolating particles 17 (such as cells, spheroids or tumors) in a liquid medium 16 in a reservoir 15 by individually aspirating 13a the particles 17 from the reservoir 15, guiding them through a channel 10 like a capillary tube, and transporting them to a target location. The device 1, for example, provides an automated platform in in vitro experiments and three-dimensional cell culture models (spheroids, organoids, e.g., microtumors), which can be used in a clinical setting, initially for the purpose of personalized treatment selection.

[0071] The device 1 is configured to isolate particles 17 present in a liquid medium 16 by a suction process 13a and transport the isolated particles, i.e., single particles 17a, through a channel 10 (like a disposable tube) so that they are directed to an output 12 of the tube. The device 1 describes a liquid handling platform, which includes: a channel 10, such as a disposable tube, as a suction opening 11 at one end and an output 12 at the other end, to which a suction force generated by a pressure generator 14 (i.e., a suction generating device) and directed in the direction of the output can be applied. The suction opening, i.e., the inlet 11, can be immersed in a reservoir 15, which is configured to contain the liquid medium 16 and the particles 17. The particles 17 are moved to the vicinity 32 of the suction opening 11 by a device, i.e., an actuator 30. The actuator 30 can be arranged in or at the reservoir 15 to generate a particle movement, i.e., a relative movement 18. Particles 17a in the vicinity 32 of the suction opening 11 can be sucked from the suction opening 11 into the tube 10 by the suction force applied to the tube 10. The sucked particles 17b can be transported through the tube 10 to the output, i.e., the outlet 12, by the microfluidic flow 21 generated by the pressure generator 14.

[0072] in other words, Figure 1 A device 1 for isolating particles 17 in an absorbent liquid medium 16 from a reservoir 15 is shown. The device 1 comprises a channel 10, like a disposable tube, having two openings, one opening as a suction input, i.e. an inlet 11, and the other opening as an output, i.e. an outlet 12. In addition, the device 1 comprises a reservoir 15 for accommodating particles 17 and liquid medium 16, and the suction opening (i.e. the inlet 11) of the channel 10 can be located in the reservoir 15. Moreover, the device can include a pressure generator 14, like a suction generating device, at the channel 10, the pressure generator 14 generates, for example, suction on demand, which sucks a single particle 17a in the vicinity 32 of the suction opening 11 to the channel 10. In addition, the device 1 comprises a moving device, i.e. an actuator 30, configured to move the particle 17 to the vicinity 32 of the suction opening 11 of the channel 10.

[0073] Device 1 may include Figures 2 to 9 Describe the features and / or functionality.

[0074] Figure 2 Shown as Figure 1 Optional additional features and / or functionalities of device 1. Figure 1 The device 1 may include one or more features and / or functions described below:

[0075] According to an embodiment, the reservoir 15 is mounted on a motor-operated reservoir table. The motor-operated reservoir table represents an actuator 30 and is configured to move the reservoir 15 so as to bring a single particle 17a into the vicinity 32 of the inlet 11 of the channel 10. Figure 2 As shown, particles 17 may be deposited on the bottom surface 15a of the reservoir 15, and the actuator 30 may be configured to move the reservoir 15 without disturbing the deposited particles 17, for example, using a speed equal to or less than 1 mm / s, 5 mm / s, 10 mm / s, or 20 mm / s and a speed equal to or less than 3 mm / s. 2 , 25mm / s 2 , 50mm / s 2 or 100mm / s 2 The acceleration is used to move the reservoir 15. Therefore, the particle 17 has a fixed position in the reservoir 15, and the actuator 30 is configured to systematically move the reservoir 15 in three dimensions so as to move the particle 17 together with the reservoir 15 relative to the inlet 11 of the channel 10.

[0076] According to an embodiment, the device 1 comprises a reservoir monitoring device 27, i.e. a monitoring device, configured to monitor the reservoir 15 or at least a monitoring area of ​​the reservoir 15 to obtain information about the position of the single particle 17a in the reservoir 15. The actuator 30 is configured to cause a relative movement to bring the single particle 17a into the vicinity 32 of the inlet 11 based on the information about the position of the single particle 17a in the reservoir 15. For example, the actuator 30 is configured to move the reservoir 15 and, with it, the deposited particles 17a relative to the inlet 11 so that the position of the single particle 17a is located within the vicinity 32 of the inlet 11. The actuator 30 can be configured to obtain the information about the position of the particle 17a directly from the reservoir monitoring device 27 and determine the necessary movement of the reservoir 15 in order to bring the single particle 17a to the inlet 11. Optionally, the device 1 may include a processor 28 configured to obtain information about the position of individual particles 17a in the reservoir 15, determine information about the necessary movement of the reservoir 15, and provide the information about the necessary movement of the reservoir 15 to the actuator 30 so that the actuator 30 can apply the necessary movement to the reservoir 15.

[0077] According to an embodiment, the reservoir monitoring device 27 is configured to obtain information about the position of particles 17 within the monitoring area of ​​the reservoir monitoring device 27. The processor 28 can be configured to determine the distance between the particles 17 located in the monitoring area based on the position of the particles 17 provided by the reservoir monitoring device 27. The processor 28 can be configured to take into account the determined distances to select a single particle 17a to be aspirated through the inlet 11. For example, the processor can be configured to check for each particle 17 in the monitoring area whether all neighboring particles have at least a predetermined distance to the respective particle 12. If this is the case for a certain particle 17, the processor 28 can be configured to select the same single particle 17a to be aspirated 13a. The processor 28 is configured to provide information about the position of the selected single particle 17a to the actuator 30.

[0078] The combination of a reservoir monitoring device 27 (eg, for reservoir volume) and a motor-operated reservoir table (ie, actuator 30) is capable of selectively moving deposited particles 17 in the reservoir 15 into proximity 32 of the suction opening (ie, inlet 11).

[0079] According to an embodiment, the reservoir monitoring device 27 or other monitoring device can be configured to monitor the vicinity 32 of the inlet 11 of the channel 10 to detect particles 17. If a single particle 17a is detected near the inlet 11, the reservoir monitoring device 27 or other monitoring device is configured to trigger the pressure generator 14, and the pressure generator 14 is configured to generate a pressure gradient in the channel 10 in response to the received trigger. Alternatively, the controller / processor 28 can be configured to control the pressure generator 14 to induce a pressure gradient in the channel 10 if a particle 17a is detected near the inlet 11 of the channel 10 by the reservoir monitoring device 27 or other monitoring device.

[0080] According to an embodiment, the reservoir monitoring device 27 or other monitoring device (like the devices described above) can be configured to obtain information about the properties of the single particle 17a (like size, material and / or shape), and about the time instant at which the single particle 17a was drawn through the inlet 11. A calculator, such as a processor 28, can be configured to determine the position of the single particle 17a within the channel 10 based on the following information:

[0081] - information about the size of the individual particles 17a,

[0082] - information about the material of the particles 17a,

[0083] - information about the shape of the particles 17a,

[0084] - information about the instant in time at which a single particle 17a is sucked through the inlet 11, and

[0085] - Information about the flow through the channel 10 .

[0086] In other words, the particle position in the channel 10 can be calculated based on the aspiration time, the tube flow rate 21 and the particle properties so that it can be determined at what time the aspirated particle 17b is located at a certain position in the channel 10. The device 1 can include a decision-based injection device 40, which is configured to determine whether the single particle 17a is present in a predetermined volume of the channel 10 located near the outlet of the channel based on the position of the single particle 17a in the channel 10. The predetermined volume, for example, represents the volume of the liquid 16 to be output next by the pressure generator 14. The decision-based injection device 40 can be configured to inject the liquid in the predetermined volume to the first target location 24b if the single particle 17a is not present in the predetermined volume, and to inject the liquid in the predetermined volume to the second target location if the single particle 17a is present in the predetermined volume.

[0087] According to an embodiment, the device 1 includes a channel volume monitoring device 26 configured to monitor a predetermined volume of the channel 10 located near the outlet 12 of the channel 10. The channel volume monitoring device 26 is configured to obtain information about whether a single particle 17a will be present in the liquid 16 to be output when the next output is performed by the pressure generator 14, for example, information about whether a single particle 17a will be present in the next droplet 22 output through the outlet 12. Optionally, the predetermined volume represents the volume of the liquid 16 to be output next time. In this case, the information about whether a single particle 17a will be present in the predetermined volume directly indicates whether a single particle 17a will be present in the liquid 16 to be output when the next output is performed by the pressure generator 14. Optionally, as Figure 2 As shown, the predetermined volume may represent the volume of the channel 10, which includes the volume of the liquid 16 to be output next time, or may represent the volume of the channel 10, which is arranged before the volume of the liquid 16 to be output next time. In this case, the channel volume monitoring device 26 or the processor 28 may be configured to determine the position of a single particle 17a within the predetermined volume at a certain time instant and calculate the time instant at which the single particle 17a will be present in the volume of liquid 16 to be output by the pressure generator 14. For example, the channel volume monitoring device 26 can provide information about the next dispensed droplet 22, for example, information about the presence or absence of a particle in the droplet 22.

[0088] According to an embodiment, if the channel volume monitoring device 26 or the processor 28 indicates that a single particle is present in the liquid to be output, in addition to the channel volume monitoring device 26, the device 1 may include a decision-based injection device 40, which is configured to inject the liquid 16 with the single particle 17a to a second target position 24b, such as a collecting substrate. Otherwise, the decision-based injection device 40 may be configured to inject the liquid 16 without the single particle 17a to the first target position 24a, that is, if the information indicates that the single particle 17a is not present in the liquid 16 to be output. The first target position 24a may also be understood as a rejection volume or a discarded volume. The decision-based injection device 40 may be configured to move a portion of the channel 10 including the outlet 12 or to move the first target position 24a and the second target position 24b so that the liquid 16 to be output is injected to the correct target position 24a or 24b through the pressure generator 14. As Figure 2As shown, it is possible that the decision-based ejection device 40 is configured to eject the liquid 16 with a single particle 17a to a location within the second target location 24b. The decision-based ejection device 40 can be configured to separate and classify droplets 22 with and without particles using vacuum extraction, motor operation stations, electromagnetic deflection devices, pneumatic deflection devices, or electrical deflection devices.

[0089] For example, the channel volume monitoring device 26 provides information about the predetermined volume of the channel 10, such as the flow rate within the predetermined volume and / or the position of the particles 17 within the predetermined volume, and / or whether a single particle 17a dispensed in a droplet 22 will be output by the pressure generator 14. The deposition location 23 of the dispensed droplet 22 can be approached by the motor-operated target substrate table 25. The decision-based ejection device 40 can be configured to control the motor-operated target substrate table 25 to eject the droplet 22 to the second target location 24b, for example, to the deposition location 23 within the second target location 24b.

[0090] According to an embodiment, the device 1 may include a channel volume monitoring device 26 and the above-described calculator for calculating the position of a single particle 17a within the channel 10 to improve the decision of the decision-based injection device 40 as to whether the liquid output through the outlet 12 must be injected into the first target position 24a or the second target position 24b.

[0091] Optionally, the device 1 may include a droplet observation device for obtaining information about the dispensed droplet 22, for example, information about the content of the dispensed droplet 22. The information about the droplet 22 may at least indicate the presence or absence of particles in the droplet volume. The decision-based ejection device 40 may be configured to move the resulting droplet 22 to a rejection position, i.e., a first target position 24a, using a motor-operated table, pneumatic or electrical deflection device in the absence of particles in the droplet 22.

[0092] The monitoring devices described herein, such as the reservoir monitoring device 27 and the channel volume monitoring device 26, can be optical observation devices, such as cameras or light absorption detectors, or impedance sensors, to observe the reservoir volume or tube volume, i.e., the predetermined channel volume. The reservoir monitoring device 27 can be located outside the reservoir 15, and the channel volume monitoring device 26 can be located outside the channel 10.

[0093] The reservoir monitoring device 27 may be configured to acquire information regarding the location, number, size, and shape of the particles 17 as well as regarding whether and when the particles 17 were aspirated.

[0094] According to an embodiment, the device comprises a controller / processor 28. Hence, the system is controlled and automated by the processor 28. This results in efficient transport of the particles 17.

[0095] According to an embodiment, the pressure generator 14 represents a droplet generation device. The pressure generator 14 is configured to output the liquid 16 in the form of droplets 22, for example, the pressure generator 14 can be configured to output a single particle 17a through the outlet 12 of the channel 10 by dispensing the single particle 17a in free-flying droplets 22. Suction can be generated by the droplet generation device, for example, sucking the liquid 16 with or without the single particle 17a through the inlet 11.

[0096] Figure 3 The pumping process 13a of the device 1 described herein is typically shown. The actuator 30, namely 301 and / or 302, is configured to move particles 17 relative to the inlet 11 of the channel 10 so as to bring a single particle 17a into the vicinity 32 of the inlet 11 to be pumped through the inlet 11.

[0097] According to a first embodiment, the actuator 30 represents a motor-operated reservoir positioner, ie a motorized positioner, 301 configured to move the reservoir 15 relative to the stationary inlet 11 of the channel 10. The movement may be performed in two dimensions or in all three dimensions.

[0098] According to a second embodiment, the actuator 30 represents a motor-operated channel positioner, ie a motorized positioner, 302 configured to move the channel 10 relative to the stationary reservoir 15. The movement may be performed in two dimensions or in all three dimensions.

[0099] According to a third and preferred embodiment, the actuator 30 includes a first actuator (e.g., a motor-operated reservoir positioner 301) and a second actuator (e.g., a motor-operated channel positioner 302) to perform relative movement between a single particle 17a in the reservoir 15 and the inlet 11 of the channel 10. The first actuator 301 can be configured to move the reservoir 15 in the x-dimension 341 and the y-dimension 342, and the second actuator 302 can be configured to move the inlet 11 of the channel in the z-dimension 343.

[0100] In one embodiment of the present invention, the reservoir 15 includes a bottom surface 15a that is large enough so that particles 17 contained in the liquid medium 16 can be deposited in a single layer on the bottom surface 15a. The actuator 30 is configured to perform relative motion so that the deposited particles 17 can be drawn through the inlet 11.

[0101] The particles 17 can be delivered to the vicinity 32 of the suction opening 11 specifically or randomly by the actuator 30. Together with the previously mentioned reservoir monitoring device 27 providing position information of the individual particles 17a, the specific delivery can be achieved. With the knowledge of the position of the individual particles 17a in the reservoir 15, the actuator 30 is configured to perform a specific relative movement to bring the individual particles 17a into the vicinity of the inlet 11.

[0102] In one embodiment of the present invention, a microfluidic chip is used as the reservoir 15 of the device 1, see Figure 4 . The particles 17 are concentrated by the specific geometry of the flow channel 185 and / or by the microfluidic flow 186 so that they are arranged at regular distances from each other. The particles 17 can be transported to the inlet 11 at regular intervals. The actuator 30 of the device 1 is configured to induce a flow 186 in the microfluidic chip to separate the particles 17 from each other and to transport the particles 17 individually to the vicinity 32 of the inlet 11 of the channel 10. The actuator 30 can represent a pump that induces the flow 186 in the microfluidic chip. In the microfluidic chip, the separation of the particles 17 can be assisted by a curved shape of microchannels (such as a liquid reservoir channel). The curved shape of the microchannels in the microfluidic chip can contain liquid 16 and particles 17. As Figure 4 As shown, the end of the microchannel is associated with the inlet 11 of the channel 10, i.e., fluidly connected, so as to enable a single particle 17a to enter the suction 13a of the channel 10 from the reservoir 15 through the inlet 11. The microfluidic chip can contain chambers and / or channels (i.e., microchannels) through which the fluid flows, wherein the chambers and / or channels have dimensions in the micrometer range or below. The microfluidic chip can include molded or engraved microchannel patterns.

[0103] Figure 5 The actuator 30 of the device 1 is shown to represent a pump configured to induce a flow 182 in the reservoir 15. Particles 17, for example, are suspended in the reservoir 15. The actuator 30 is configured to transport the particles 17 separately, for example randomly, to the inlet 11. The random particle motion can be induced, for example, by a pump, such as a syringe pump, a roller pump, etc. Figure 5 , the pump is configured to induce a mixing flow 182 in the reservoir 15. For example, the actuator 30 is configured to induce the flow 182 to lift the deposited particles 17 to the inlet 11 immersed in the reservoir 15. The actuator 30 can be configured to induce the flow 182 to separate the particles 17 within the reservoir, for example, the flow 182 can prevent the aggregation or accumulation of the particles 17, especially near the inlet 11 32.

[0104] Likewise, a mixing flow 182 may be induced in the reservoir 15 by paramagnetic particles 183. The actuator 30 of the apparatus 1 may include a permanent magnet 184 disposed outside the reservoir 15 and paramagnetic particles 183 disposed inside the reservoir 15. The actuator 30 is configured to induce a stirring motion of the paramagnetic particles 183 inside the liquid 16 in the reservoir 15 by the permanent magnet 184, see Figure 6 The flow 182 induced by the actuator 30 lifts or transports the particles 17 to the inlet 11, e.g., as in Figure 5 , for a description of stream 182.

[0105] If device 1 includes a Figure 5 and Figure 6 If one of the actuators 30 described is included in the device 1, the device 1 may further include a particle concentration monitoring device, which is configured to determine the particle concentration of the liquid 16 containing the particles 17 in the reservoir 15. A particle concentration equal to or below a predetermined threshold increases the probability of aspirating a single particle 17 through the inlet. A particle concentration above a predetermined threshold increases the possibility of aspirating more than one particle 17 at the same time, or more precisely allows the aspiration of two or more particles close to each other. Therefore, it is desirable that the particle concentration in the reservoir 15 is below the predetermined threshold so that a single particle is aspirated at a certain time interval. For example, the pressure generator 14 can be configured to output a certain amount of liquid 16 at each time instant, and the predetermined threshold defines a particle concentration at which only one particle 17 is present in the certain amount of liquid 16 output by the pressure generator 14 at each time instant. The particle concentration monitoring device may be configured to indicate whether the particle concentration exceeds a predetermined threshold, or may be configured to automatically adjust the particle concentration by adding further liquid 16 to the reservoir 15 to reduce the particle concentration if the particle concentration exceeds a predetermined threshold.

[0106] In one embodiment of the present invention, Figure 7As shown, suction is applied to the tube output, i.e., the outlet 12, and the suction can be generated by a droplet generating device. The droplet generating device can represent the pressure generator 14 of the device 1 described herein. For example, a piezoelectrically driven plunger 141 can be used, which dispenses droplets 22 from the tube output 12 by actuation on the tube (i.e., the channel 10). The device 1 may include a flexible or deformable channel 1. The plunger 141 of the pressure generator 14 is configured to deform the channel 10 near the outlet 12 to output a certain amount of liquid 16 with or without individual particles 17a, for example, in the form of droplets 22. Moreover, the plunger 141 of the pressure generator 14 is configured to stop the deformation of the channel 10 near the outlet 12 to induce a pressure gradient in the channel 10 and draw the liquid 16 with or without individual particles 17a through the inlet 11 of the channel 10. Alternatively, a thermal droplet generating device can be used as the pressure generator 14.

[0107] Figure 8 and Fig. 9 A further option of the pressure generator 14 of the device 1 described herein is shown. Both embodiments have in common that the outlet 12 and the pressure generator 14 communicate with the collection chamber 120. The pressure generator 14 can be configured to generate a negative pressure in the collection chamber 120 and thus induce a pressure gradient within the channel 10 for the suction process 13a through the inlet 11 of the channel 10, for the transport 21 through the channel 10 and for the output 13b through the outlet 12 of the channel 10. The suction force 50 can be generated by the negative pressure or the positive pressure induced by the pressure generator 14. Figure 8 and Fig. 9 The pressure generator 14 in the embodiment can be realized as a pump, like a syringe pump, a roller pump, etc. For example, the pump can be configured to generate a negative pressure and thus a suction force 50 in the collection container (i.e. the collection chamber 120), which transports the sucked particles 17b into the collection container 120. This type of configuration of the invention can be used to discharge a clearly defined amount of particles 17 from a particle suspension very easily and quickly.

[0108] Figure 8 and Fig. 9 The embodiment shown in FIG. 1 differs in that Fig. 9 The collection container 120 also includes a functional structure 121 to further classify or manipulate the particles 17.

[0109] Fig.10An embodiment of a method 100 for isolating particles is shown. The method 100 includes fluidly coupling an inlet 110 of a channel with a reservoir holding a liquid containing particles. In addition, the method 100 includes causing 120 relative motion between the particles and the inlet of the channel so as to bring a single particle to the vicinity of the inlet to be drawn through the inlet. Furthermore, the method 100 includes generating 130 a pressure gradient in the channel by which the liquid containing the single particle is drawn from the reservoir against gravity through the inlet of the channel and outputting 140 the liquid through the outlet of the channel.

[0110] According to an embodiment, the method 100 includes causing 120 relative motion by moving the reservoir and / or the inlet of the channel so as to bring a single particle (preferably a sedimentation particle) into proximity of the inlet to be drawn through the inlet. According to a preferred embodiment, the method 100 performs causing 120 relative motion by moving the reservoir in two dimensions and moving the channel in a third dimension perpendicular to the plane defined by the two dimensions.

[0111] According to an embodiment, the method 100 includes causing 120 a relative motion by inducing a mixed liquid flow in the reservoir to prevent the particles from being deposited in the reservoir and to bring the particles individually into the vicinity of the inlet of the channel.

[0112] According to an embodiment, the reservoir is a reservoir channel within a microchip, such as a microchannel, and method 100 includes fluidically connecting an inlet 110 to an end of the reservoir channel and causing 120 relative movement by causing flow of liquid in the reservoir channel to bring particles individually into proximity of the inlet of the channel.

[0113] According to an embodiment, the method 100 comprises monitoring the reservoir to obtain information about the position of a single particle in the reservoir, and causing 120 a relative movement to bring the single particle into the vicinity of the inlet based on the information about the position of the single particle.

[0114] According to an embodiment, the method 100 includes monitoring a vicinity of an inlet of a channel, if a particle is detected in the vicinity of the inlet of the channel, and generating 130 a pressure gradient in the channel.

[0115] According to an embodiment, method 100 includes acquiring information about the size and / or shape of a single particle and information about the instant in time when the single particle is drawn through an inlet, and determining the position of the single particle within the channel based on the acquired information and based on information about the flow velocity through the channel.

[0116] According to an embodiment, method 100 includes determining information about whether the single particle exists at an end near an outlet of the channel based on a determined position of the single particle within the channel, and based on this information, spraying liquid without the single particle to a first target position or liquid with the single particle to a second target position.

[0117] According to an embodiment, the method 100 includes monitoring a predetermined volume of the channel located near an outlet of the channel and obtaining information about whether a single particle is present in the predetermined volume. In addition, the method 100 may include, if the information indicates that a single particle is not present in the liquid to be output, spraying the liquid without the single particle to a first target location; if the information indicates that a single particle is present in the liquid to be output, spraying the liquid with the single particle to a second target location.

[0118] According to an embodiment, method 100 includes determining a particle concentration in a fluid containing particles in a reservoir and indicating whether the particle concentration exceeds a predetermined threshold.

[0119] According to an embodiment, method 100 includes outputting 140 the single particle through an outlet of a channel by dispensing the single particle in a free-flying droplet.

[0120] An embodiment relates to a method 100 of selectively or randomly drawing one or more particles through a disposable tube by a suction generating device, i.e., a pressure generator, comprising the steps of:

[0121] - Fill the reservoir with the particle suspension and immerse the suction opening (ie inlet) of the tube (ie channel) into it.

[0122] - For selective aspiration (active): Select the appropriate particles with the reservoir monitoring device. In particular, move the appropriate particles to the vicinity of the aspiration opening of a channel (e.g. disposable tube).

[0123] - For random suction (passive): Move the medium to keep the particles in suspension. The fluid movement brings the particles randomly into the vicinity of the suction opening of the tube.

[0124] - Using a reservoir observation device, ie a reservoir monitoring device, information about particles in the reservoir in the vicinity of the opening (ie the inlet) of the aspiration tube is acquired.

[0125] - Generate suction at the suction opening of the tube using a pressure generator, such as a suction generating device as required, and suck the particles in the vicinity of the suction opening into the tube.

[0126] - Calculate the particle position in the tube based on particle size, shape and material, pumping time and tube flow rate using information collected by the reservoir monitoring device to determine when the pumped particles reach the output of the tube, i.e. the outlet.

[0127] - The aspirated particles are transported through the disposable tube by a flow generated by a suction generating device (ie a pressure generator).

[0128] Although some aspects have been described in the context of an apparatus, it is clear that these aspects also represent a description of a corresponding method, where a module or device corresponds to a method step or a feature of a method step. Similarly, aspects described in the context of a method step also represent a description of a corresponding module or item or feature of a corresponding apparatus.

[0129] The embodiments described above are merely illustrative of the principles of the present invention. It is understood that modifications and variations of the details and arrangements described herein will be apparent to other persons skilled in the art. Therefore, herein, it is intended to be limited only by the scope of the pending patent claims, and not by the specific details present in the manner of description and explanation of the embodiments.

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Claims

1. An apparatus for particle isolation (1), comprising: a liquid reservoir (15) for containing a liquid (16) containing particles (17); a channel (10) having an inlet (11) and an outlet (12), wherein the inlet (11) is configured to be fluidically coupled to the reservoir (15) containing the liquid (16) containing particles (17); The pressure generator (14) is configured as follows: a pressure gradient is generated in the channel (10) by which a liquid (16) containing a single particle (17a) is drawn (13a) from the reservoir (15) against gravity (2) through the inlet (11) of the channel (10), and causing liquid (16) to be output through the outlet (12) of the channel (10); An actuator (30) is configured to induce relative motion between the particle (17) and the inlet (11) of the channel (10) so as to cause a single particle (17a) in the vicinity (32) of the inlet (11) to be sucked (13a) through the inlet (11).

2. Apparatus (1) according to claim 1, wherein the actuator (30) is configured to move the reservoir (15) and / or the inlet (11) of the channel (10) so as to bring the single particle (17a), preferably the deposited particle, to the vicinity (32) of the inlet (11) to be sucked (13a) through the inlet (11).

3. The device (1) according to claim 1, wherein the actuator (30) is configured to induce a mixed liquid flow (182) in the liquid reservoir (15) to prevent particles (17) from being deposited in the liquid reservoir (15) and to bring the particles (17) individually into the vicinity (32) of the inlet (11) of the channel (10).

4. The device (1) according to claim 1, wherein the reservoir (15) is a reservoir channel (185) within a microchip, the inlet (11) is fluidically connected to the end of the reservoir channel (185), and wherein the actuator (30) is configured to cause a flow of the liquid (16) in the reservoir channel (185) to bring the particles (17) individually into the vicinity (32) of the inlet (11) of the channel (10).

5. The device (1) according to one of claims 1 to 4, wherein the device (1) comprises, a reservoir monitoring device (27) configured to monitor the reservoir (15) to obtain information about the position of the individual particles (17a) in the reservoir (15), and wherein the actuator (30) is configured to cause relative movement to bring the single particle (17a) into the vicinity (32) of the inlet (11) based on the information about the position of the single particle (17a).

6. The device (1) according to claim 5, in, The reservoir monitoring device (27) is configured to obtain information about the location of particles (17) located in a monitoring area of ​​the reservoir monitoring device (27); The device (1) further comprises a processor (28), wherein the processor is configured to: determining a distance between the particles (17) located in the monitoring area based on the information about the positions of the particles (17) located in the monitoring area, selecting the single particle (17a) to be aspirated from the particles (17) located in the monitoring area based on the determined distances between the particles (17), and The information about the position of the single particle (17a) to be aspirated is provided to the actuator (30).

7. The device (1) according to one of claims 1 to 6, wherein the device (1) comprises, a monitoring device (27) configured to monitor the vicinity (32) of the entrance (11) of the passage (10), and A controller (28) is configured to control the pressure generator (14) to induce the pressure gradient in the channel (10) if particles are detected near (32) the inlet (11) of the channel (10) by the monitoring device.

8. The device (1) according to one of claims 1 to 7, wherein the device (1) comprises, a monitoring device (27) or the monitoring device (27) configured to acquire information about the size and / or shape of the single particle (17a) and about the instant in time at which the single particle (17a) is drawn (13a) through the inlet (11), and A calculator (28) is configured to determine the position of the single particle (17a) within the channel (10) based on the information obtained by the monitoring device and based on information about the flow velocity through the channel (10).

9. The device (1) according to claim 8, wherein the device (1) comprises a decision-based injection device (40), the decision-based injection device (40) being configured to: determining information as to whether the single particle (17a) is present in a predetermined volume of the channel (10) located near the outlet (12) of the channel (10) based on the position of the single particle (17a) within the channel (10) obtained from the calculator (28), and Based on this information, the liquid (16) without the individual particles (17a) is sprayed to a first target location (24a) or the liquid (16) with the individual particles (17a) is sprayed to a second target location (24b).

10. The device (1) according to one of claims 1 to 9, wherein the device (1) comprises, A channel volume monitoring device (26) is configured to: monitoring a predetermined volume of the channel (10) located proximate the outlet (12) of the channel (10), and obtaining information about whether the single particle (17a) is present in the predetermined volume, and a decision-based injection device (40) or the decision-based injection device (40) configured to, If the information indicates that the single particle (17a) is not present in the liquid (16) to be delivered, ejecting the liquid (16) without the single particle (17a) to a first target location (24a), and If the information indicates that the single particle (17a) is present in the liquid (16) to be output, the liquid (16) with the single particle (17a) is sprayed to a second target location (24b).

11. The device (1) according to one of claims 1 to 10, wherein the device (1) comprises a particle concentration monitoring device, the particle concentration monitoring device being configured to: determining a particle concentration in the liquid (16) containing the particles (17) in the reservoir (15), and Indicates whether the particle concentration exceeds a predetermined threshold.

12. Device (1) according to one of claims 1 to 11, wherein the channel (10) is a continuous filter-free channel.

13. The device (1) according to one of claims 1 to 12, wherein the channel (10) is a hose having a uniform diameter over its entire length.

14. Apparatus (1) according to one of claims 1 to 13, wherein the pressure generator (14) is configured to output the single particle (17a) through the outlet (12) of the channel (10) by dispensing the single particle (17a) in free-flying droplets.

15. A method (100) for particle isolation, comprising the steps of: fluidly coupling an inlet of the channel (110) to a reservoir holding a liquid containing particles; causing (120) relative motion between the particle and the inlet of the channel to bring a single particle into proximity with the inlet to be drawn through the inlet; creating (130) a pressure gradient in the channel, whereby liquid containing the single particle is drawn from the reservoir through the inlet of the channel against gravity by the pressure gradient; Liquid is output (140) through an outlet of the channel.

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