Sampling device

Abstract

A sampling device (100) comprising a probe (220), the probe comprising: a first lumen (222) configured to dispense a first fluid at a distal end (230) of the probe; a second lumen (223) configured to aspirate the first fluid at the distal end (230) of the probe, wherein a distal end of the first lumen is adjacent to a distal end of the second lumen, wherein the sampling device (100) is configured to dispense the first fluid via the first lumen (322) and simultaneously aspirate the first fluid via the second lumen (323).

Description

Technical Field

[0001] This invention relates to an apparatus and method for sampling biological material. In particular, but not exclusively, this invention relates to an apparatus and method for endoscopic sampling. Background Technology

[0002] Pulmonary epithelial lining fluid (ELF) is an aqueous layer covering the surface of the lungs, composed of phospholipids, proteins, and surfactants, which promote healthy lung function and optimal gas exchange. ELF acts as a barrier against irritants and pathogens. Furthermore, ELF enhances mucociliary clearance and plays a role in the immune response. ELF is distributed throughout the conduction airways and alveolar regions of the lungs.

[0003] ELF contains a variety of cellular and molecular components that can provide information about lung diseases and infections. ELF reveals information about the penetration of inhaled ingredients and anti-infective agents, which is crucial for monitoring inhaled medications and simulating lung deposition.

[0004] Cytological analysis of ELF cell components in airway secretions can be used for differential diagnosis of diseases; however, much work remains to be done to understand the distribution of different cell populations throughout the lung regions of the bronchial tree. Such work depends on the capabilities of existing sampling tools.

[0005] Proteomic analysis of ELF can assess protein abundance and provide information about protein function and expression in the lungs, thus contributing to a better understanding of basic lung biology and disease. It can also provide information about the efficacy of gene therapy by demonstrating evidence of transgenic expression delivered in protein secretions. A fundamental challenge is detecting low-abundance protein targets in ELF.

[0006] When ELF is sampled using any of the methods described below, the ELF will be diluted in the fluid used for sampling or elution. This necessitates an accurate estimation of the ELF concentration in the sample fluid to provide useful conclusions for downstream analysis.

[0007] The dilution factor of ELF in a sample is estimated using small, endogenous molecules that readily cross the blood-bronchial barrier. This is based on the assumption that these molecules are substantially at the same level in serum and ELF due to their free movement across the barrier. The relative levels of albumin and urea in the sample fluid, as well as in serum, have been used to estimate the dilution factor of ELF in the sample. In some cases, albumin provides a more accurate and reasonable estimate than urea; however, in patients with lung disease, albumin is significantly elevated and exhibits considerable variability, making it unsuitable as a reference.

[0008] Alternative exogenous markers (such as technetium-99m diethylenetriaminepentaacetic acid) 99m Tc-DTPA or isotonic mannitol can provide more reliable ELF concentration estimates, but the process is time-consuming and requires intravenous injection and infusion of reference molecules.

[0009] Endogenous labeling is preferred, and urea-based methods are commonly employed. However, due to the high mobility of urea, it has been observed to diffuse significantly from areas outside the sampling region, both in blood and surrounding sites. Furthermore, the diffusion of urea into the sample area is significantly influenced by the state of lung surfactant in ELF, making accurate assessment difficult. It is generally believed that urea levels may be artificially inflated, leading to an overestimation of ELF concentration in the sample.

[0010] To minimize the overestimation of ELF content, care must be taken to minimize the volume ratio of sample fluid to ELF (error is proportional to volume), ensure that the relative osmotic pressure of the irrigation fluid and serum is the same (and minimize the residence time of the wash, typically within one minute).

[0011] Lung fluid sampling methods are generally classified into blind methods and guided methods.

[0012] Blind lung sampling method

[0013] Blind lung fluid sampling encompasses a variety of non-bronchoscopic methods, including mini-broncho-alveolar lavage (BAL), first reported in the 1980s, non-bronchial NB BAL, s-Cath / s-BAL, or protected BAL. Non-bronchoscopic sampling involves blindly advancing a suction catheter into the distal airway, where it is embedded to enable edema fluid sampling (s-cath) or lavage (mini-BAL / NB-BAL / protected BAL). Protected BAL minimizes contamination from the upper respiratory tract by employing an endotracheal catheter wedged into the airway and through which the sampling catheter is introduced; however, the primary sampling mechanism for performing low-volume BAL via a single-lumen sampling catheter is consistent across the methods. These methods have reportedly similar sensitivity and specificity to bronchoscopic BAL. Chest X-ray showed catheter embedding in the right bronchial tree. However, some worry that this blind sampling method could lead to false negatives if samples are taken from unaffected areas of the lungs.

[0014] The blind approach was developed for patients deemed unsuitable for bronchoscopic arthroscopy (BAL) and best suited for the least invasive procedure available. These patients include critically ill patients, mechanically ventilated patients, immunocompromised patients, or children in whom bronchoscopic arthroscopy (BAL) cannot be performed using an endotracheal tube with an internal diameter <4.5 mm (i.e., infants) or where bronchoscopic facilities for BAL are not available at all.

[0015] Commercially available miniature BAL sampling catheters are readily available, and their relatively rapid sampling (compared to guided BAL) has been used to guide rational antibiotic selection, thereby reducing unnecessary antibiotic use in the rapid response to infections in critically ill patients.

[0016] Blind methods have not been standardized and vary depending on catheter type, catheter location, and the volume of fluid being infused.

[0017] Guided lung fluid sampling method

[0018] Bronchoalveolar lavage (BAL)

[0019] BAL (Bronchoalveolar Assistance) via flexible bronchoscope was first reported and introduced in the 1970s and is currently the standard diagnostic tool for lung infections and diseases. BAL is currently the clinical gold standard for lung fluid sampling.

[0020] BAL involves introducing a large amount of fluid into the lungs, which is then removed by aspiration. The sample is captured in the fluid for downstream analysis. Reviews of the diagnostic and therapeutic applications of BAL exist in the literature.

[0021] BAL is performed after bronchoscopy to guide sampling. This procedure is not standardized and varies depending on the application / location, but the general principle involves advancing the bronchoscope until it is wedged into the distal end, thereby blocking the airway leading to the target area.

[0022] Due to poor recovery rates, multiple saline flushes are typically performed and the saline solution is collected (up to approximately 200 mL) by wedging the bronchoscope into the airway and dispensing / aspirating saline solution through the biopsy port of the endoscope.

[0023] BAL is well tolerated in healthy patients; however, several complications have been reported, including lung and airway irritation, transient hypoxemia, fever (in approximately 30% of patients), bronchospasm, acute respiratory distress syndrome, and, in rare cases, pneumothorax. False positives are common, and several methodological issues need to be considered when assessing the clinical relevance of the obtained samples, suggesting that BAL should always be used in conjunction with comprehensive clinical information in any diagnosis.

[0024] The volume of ELF recovered from BAL is relatively small compared to the infused volume; reportedly, as little as 2 mL of ELF has been recovered from a collected 200 mL BAL fluid (divided into four 50 mL lavages). In this case, the residence time is approximately 2 minutes, suggesting that ELF levels may be inflated due to dilution estimation errors. The low recovery rate of ELF in BAL fluid and the inability to perform localized sampling have spurred the development of other, more targeted ELF sampling methods. BAL is currently the only method capable of sampling alveolar regions.

[0025] Bronchoscopic microsampling (BMS)

[0026] Bronchoscopic microsampling (BMS) is a method in which bronchial epithelial lining fluid is collected through a bronchoscope using a device consisting of a metal wire with an adsorbent probe at the end.

[0027] Microsampling probes offer higher sensitivity and localization capabilities, enabling basic research in areas where BAL sensitivity is insufficient (such as COVID-19). However, adsorption-based microsampling methods are limited in sampling capacity, introduce elution sample dilution / contamination, and are prone to tissue damage. Because the true volume of collected ELF is estimated, and the elution and sampling steps are separate, this method makes it difficult to estimate sample dilution. Furthermore, probes cannot obtain cytological information.

[0028] Lung Microdialysis - Lung MD

[0029] Microdialysis was first described in the 1950s, but it began to be used in animals in the 1960s to study tissue biochemistry and was applied to humans in the 1990s. Microdialysis enables continuous sampling, providing high-quality information for pharmacokinetic studies and many other applications under constant perfusion conditions, including studies on calibration methods and studies specifically for pulmonary applications.

[0030] Microdialysis probes mimic capillaries, facilitating substance exchange via extracellular fluid. The dual-lumen probe is slowly perfused with a physiological fluid called the perfusion solution. Substances in the fluid surrounding the probe are perfused into the perfusion solution through a semi-permeable membrane at the probe tip and collected as dialysate at the system's outlet for downstream analysis.

[0031] Early attempts at minimally invasive probe implantation through the chest wall often resulted in pneumothorax. Tracheotomy minimizes the risk of lung collapse, and ELF measurements are obtained using microdialysis probes placed in pigs via tracheotomy. However, performing tracheotomy in healthy humans as part of trials is unethical. This created a need for surgical implantation, which limited early human microdialysis studies to patients already scheduled for thoracic surgery. Surgically implanted probes have been used in human antibiotic penetration studies and clinical trials to observe ELF cytokines during repair of open aortic aneurysms.

[0032] Bronchial microdialysis (BMD)

[0033] The limitation of requiring tracheotomy or thoracic surgery for implantation has driven the development of bronchoscopic microdialysis (BMD). Current BMD probes are typically custom variants of commercially available microdialysis catheter probes.

[0034] However, semipermeable membranes only allow analytes with <100 kDA to enter the dialysate, thus making it impossible to collect cytological data. Furthermore, the limited sample capacity at the probe tip (approximately 1.4 μL) and the underlying diffusion mechanism requiring slow sampling rates (1–2 μL / min) and several hours of perfusion prior to sampling may limit its application outside of continuous monitoring scenarios.

[0035] Numerous devices exist, such as endoscopes, that combine catheters for injecting fluids and catheters for removing fluids, as disclosed, for example, in US5823940 (Newman), US4750902 (Wuchinich), US2006 / 1073244 (Boulais), or US2023 / 0190078 (Clayman). However, these devices are not configured to sample biological samples (e.g., lung epithelial lining fluid (ELF)) from a subject for analysis.

[0036] The purpose of this invention is to solve and / or mitigate one or more problems associated with the prior art.

[0037] One object of the present invention is to provide an apparatus and / or method that enables improved local fluid biopsy in a subject. In particular, it should be understood that although the sampling of pulmonary epithelial lining fluid (ELF) has been discussed in the background art to provide background information, the present invention is not limited to ELF sampling and can be applied to any local fluid biopsy in a subject.

[0038] One object of the present invention is to provide an apparatus and / or method that enables continuous endoscopic (e.g., bronchoscopic) sampling with minimal damage to the subject's tissues, using a smaller sampling volume, and increased flexibility. Summary of the Invention

[0039] This invention is based on the discovery that a multi-lumen device can be used to perform continuous or semi-continuous endoscopic sampling simultaneously by dispensing fluid via a dispensing lumen and aspirating via different lumens. By controlling one or more parameters of the multi-lumen device (such as flow rate and / or lumen size), a continuous and / or self-sustaining fluid interface can be formed between the distal end of the multi-lumen device and the target sample, and a continuous fluid flow can be generated between the dispensing lumen and the aspiration lumen. This allows for continuous sampling with soft contact (fluid contact) with the target sample, thereby minimizing potential damage to the subject's tissues. Furthermore, the outer diameter (OD) of the device of this invention is small enough to be compatible with the biopsy ports of commercially available endoscopes, thus allowing deployment at the target site via standard lumens in conventional endoscopes. This ensures that the distal end of the device is not contaminated by contact with other parts of the subject's tissues before reaching the target sample site.

[0040] According to a first aspect, a sampling device including a probe is provided, wherein the probe includes:

[0041] A first lumen, the first lumen being configured to dispense a first fluid at its distal end;

[0042] A second lumen, configured to aspirate the first fluid at its distal end, wherein the distal end of the first lumen is adjacent to the distal end of the second lumen, wherein the sampling device is configured to dispense the first fluid via the first lumen and simultaneously aspirate the first fluid via the second lumen.

[0043] Advantageously, this allows for continuous sampling, for example, within an endoscope (e.g., within a bronchoscope). Furthermore, by controlling one or more parameters of the sampling device, such as flow rate and / or the dimensions of the first and second lumens, a self-sustaining volume of fluid can be generated at the distal end of the sampling device, allowing for simultaneous and / or continuous sampling with soft contact (fluid contact) with the target, thereby minimizing potential damage to the subject's tissues. Specifically, by controlling one or more parameters of the sampling device, such as flow rate and / or the dimensions of the first and second lumens, a self-sustaining fluid flow can be generated at the distal end of the sampling device between the first (dispensing) lumen and the second (aspiration) lumen, allowing for simultaneous and / or continuous sampling with soft contact (fluid contact) with the target, thereby minimizing potential damage to the subject's tissues.

[0044] Advantageously, the device of the present invention also allows the device to be translated during sampling, thereby enabling the normal functioning of the target (tissue), such as breathing. This contrasts with other conventional sampling methods, such as BAL, which involve wedging an endoscope into the airway and introducing a large amount of fluid into the lungs, followed by removal of the fluid by suction, thus hindering breathing during the sampling procedure.

[0045] Advantageously, the device of the present invention can also allow sampling of fluids and dried analytes.

[0046] The term "far side" will be understood herein as the position or end closest to the sampling location and / or furthest from the user or operator. Conversely, the term "proximal side" will be understood herein as the position or end opposite to the sampling location and / or closest to the user or operator.

[0047] The device can be configured to generate a self-sustaining fluid flow between the first (dispensing) lumen and the second (aspiration) lumen at its distal end. Therefore, the device can be configured to generate, during use, a volume of first fluid at or near the distal end of the probe (e.g., the distal end of the first lumen and / or the second lumen), sufficient to create a continuous fluid interface between the probe and the target (e.g., tissue of a subject).

[0048] As will be understood, the second lumen may be able to aspirate the first fluid before establishing a fluid interface with the target. In other words, the fluid aspirated by the second lumen may be the first fluid distributed via the first lumen before establishing a fluid interface with the target, thereby creating a self-sustaining and / or continuous fluid flow at its distal end between the first (distribution) lumen and the second (aspiration) lumen.

[0049] In use, when the device is deployed at or near a target, and / or when a fluid interface is established with the target, the first fluid “wets” the target and / or interacts with the target to generate a second fluid. In such a case, the second lumen is configured to aspirate the second fluid at its distal end. In other words, when a fluid interface is established between the first fluid and the target, the fluid aspirated by the second lumen can be a second fluid generated via the interaction between the first fluid and the target sample. Therefore, the second fluid can be a mixture of the first fluid and the target sample.

[0050] Typically, the probe may include a first lumen configured to dispense a first fluid at its distal end.

[0051] Alternatively, the probe may include a plurality of first lumens configured to dispense a first fluid at its distal end.

[0052] Typically, the probe may include a second lumen configured to aspirate the first fluid or the second fluid at its distal end.

[0053] Alternatively, the probe may include a plurality of second lumens configured to aspirate the first fluid or the second fluid at their distal ends.

[0054] Typically, the probe may include a first lumen and a second lumen, the first lumen being configured to dispense a first fluid at its distal end, and the second lumen being configured to aspirate either the first fluid or the second fluid at its distal end.

[0055] The device may be configured to dispense the first fluid at its distal end via the first lumen for at least a short time (e.g., at least 1 ms, at least 10 ms, at least 0.1 s, at least 1 s), and simultaneously aspirate the first fluid or the second fluid at its distal end via the second lumen.

[0056] Advantageously, before establishing a fluid interface with the target, the device can be configured to generate a continuous and / or self-sustaining (e.g., substantially constant or stable) first fluid flow between the first (dispensing) lumen and the second (suction) lumen at its distal end.

[0057] Advantageously, after establishing a fluid interface with the target, the device can be configured to generate, in use, a continuous and / or self-sustaining (e.g., substantially constant or stable) volume of a second fluid at or near the distal end of the first lumen and / or at or near the distal end of the second lumen, and / or the device can be configured to generate, in use, a continuous and / or self-sustaining (e.g., substantially constant) fluid flow between the first (dispensing) lumen and the second (suction) lumen at its distal end.

[0058] Therefore, although the device is capable of generating a self-sustaining volume of fluid at its distal end, it should be understood that when the first fluid comes into contact with the target / the target (e.g., tissue), surface tension will cause the first fluid to "wet" the target (tissue) to create a fluid interface, preferably a continuous and / or self-sustaining fluid interface. In other words, the fluid flow between the first (dispensing) lumen and the second (aspiration) lumen allows for the generation of a continuous fluid interface between the probe and the target (e.g., the subject's tissue) during use, thereby enabling continuous sampling of the target at or near the fluid interface.

[0059] The sampling device can be configured to continuously dispense a first fluid at its distal end via the first lumen, and can be configured to continuously aspirate the first fluid or the second fluid at its distal end via the second lumen.

[0060] The term "continuous" here will be understood to mean that the distribution and / or suction of the fluid is uninterrupted. However, the term may include one or more types of flow, such as steady-state (substantially constant velocity) flow, flow with a ramp-like change in velocity between different values ​​over a period of time, square wave flow, oscillating flow, pulsating flow, etc.

[0061] The sampling device can be configured to semi-continuously distribute a first fluid at its distal end via the first lumen, and / or aspirate the first fluid or the second fluid at its distal end via the second lumen. The term "semi-continuously" will be understood herein to mean that the fluid distribution and / or fluid aspiration are not necessarily strictly continuous, but can be intermittent. The intermittent flow itself can be stable or unstable, such as ramp flow, square wave flow, oscillating flow, pulsating flow, etc.

[0062] In one embodiment, the sampling device may be configured to continuously aspirate the first fluid at its distal end via the second lumen and intermittently dispense the first fluid at its distal end via the first lumen. It has been found that dispensing the first fluid intermittently does not adversely affect the fluid interface between the probe and the target (tissue), but rather can promote mixing at the interface. However, it should be understood that, as stated above, the intermittent nature of the flow should still achieve or maintain a continuous or self-sustaining fluid interface between the device (e.g., the tip of the probe) and the target. This can be achieved by stopping dispensing and / or aspiration for a short period (typically within a few microseconds). However, it should be understood that the duration of the intermittent period will depend on the specific system used and associated conditions, including, for example, probe size, target wettability, the presence of surfactants, the properties of the first fluid, the properties of the target, flow parameters, temperature, etc.

[0063] Preferably, the second lumen can be disposed adjacent to the first lumen.

[0064] The first lumen may have a proximal end and a distal end. The distal end may be located at the sampling end of the first lumen. The proximal end may be located at the end opposite to the distal end or the sampling end. The distal end of the first lumen may define a dispensing end and / or may be configured to dispense the first fluid.

[0065] The second lumen may have a proximal end and a distal end. The distal end of the second lumen may be located at the sampling end of the second lumen. The proximal end of the second lumen may be located at the end opposite to the distal end or sampling end of the second lumen. The distal end of the second lumen may define an inlet end and / or may be configured to aspirate the first fluid or the second fluid.

[0066] The first lumen may be defined by a first conduit (e.g., a first tube).

[0067] The second lumen may be defined by a second conduit (e.g., a second tube).

[0068] Preferably, the first catheter and the second catheter can be in contact with each other at least at their distal ends.

[0069] Preferably, the first catheter and the second catheter may be arranged side by side at least at their distal ends.

[0070] The device (e.g., a probe) may include a sheath.

[0071] Typically, the first lumen and the second lumen can be disposed within the sheath.

[0072] Typically, the first catheter and the second catheter can be disposed within the sheath.

[0073] The first catheter and the second catheter can contact each other within the sheath. The first catheter and the second catheter can be arranged side by side within the sheath.

[0074] The sheath may have a distal end located at or near the sampling end of the device, and a proximal end opposite the distal end of the sheath.

[0075] The width of the sheath (e.g., internal width or internal diameter) can be about 0.5-5 mm, for example, about 1-3 mm, typically about 1-1.7 mm. The width of the sheath (e.g., external width or external diameter) can be about 0.5-5 mm, for example, about 1-3 mm, typically about 1-1.9 mm or 1-1.8 mm, for example, about 1.8 mm. Suitably, the external width or external diameter of the sheath can be substantially equal to or smaller than the internal diameter of the port or internal catheter of a standard endoscope (e.g., a bronchoscope) (typically an internal diameter of about 2 mm), which allows the device of the present invention (e.g., a probe) to be placed within or compatible with conventional endoscopes (e.g., bronchoscopes). This allows the device (e.g., a probe) to be inserted into a conventional endoscope to deploy the distal end of the device to the sampling site. Advantageously, this arrangement prevents contamination of the distal end of the device, for example, due to contact with the tissue surface of the subject. This ensures that the distal end of the device (e.g., a probe) remains sterile and / or uncontaminated until it is deployed at the sample site via the distal end of the endoscope.

[0076] The width of the sheath (e.g., outer width or outer diameter) can be about 1-1.8 mm or 1-1.9 mm, for example about 1.8 mm, so as to provide a certain degree of clearance between its outer surface and the endoscope / the port of the endoscope / the port or the internal catheter / the inner surface of the internal catheter.

[0077] The wall thickness of the sheath can be less than or equal to about 300 μm, typically less than or equal to about 250 μm. This configuration allows the sampling device (e.g., a probe) to be sufficiently flexible for easy deployment to the sampling site. For example, when deployed via an endoscope, the sampling device can be sufficiently flexible for deployment via the endoscope's port and / or internal catheter.

[0078] Advantageously, the distal end of the first lumen may be adjacent to and / or substantially flush with or level with the distal end of the second lumen. In other words, the distal ends of the first and second lumens may be substantially flush with or level with each other, and / or the first and second lumens may have substantially equal z-heights relative to the proximal end of the probe. This can facilitate the formation of a constant or stable volume of first fluid at or near the end of the probe during use.

[0079] As described above, prior to establishing a fluid interface with the target, the device can be configured to generate a continuous and / or self-sustaining (e.g., substantially constant or stable) first fluid flow between the first (dispensing) lumen and the second (suction) lumen at its distal end. This can create a substantially constant or stable volume of first fluid at the distal end or tip of the probe, which in some embodiments may define or form droplets. The volume of the first fluid and / or droplets can be about 0-200 μl, for example, about 1-200 μl, for example, about 10-200 μl. The volume of the first fluid and / or droplets can be less than about 200 μl, typically less than about 183 μl. It should be understood that the precise volume of the first fluid and / or droplets may depend on many factors, such as lumen / conduit diameter, relative flow rate, first fluid viscosity, etc.

[0080] As mentioned above, it should be understood that the presence of visible droplets at or near the distal end of the probe may not necessarily be required. For example, as described above, a fluid interface can be established between the distal end of the probe and the target (tissue) when the fluid at the distal end of the probe is in brief contact with the target (tissue), and this fluid interface can be maintained, for example, due to surface tension, when the distal end of the probe is no longer in physical contact with the target (tissue) but is very close to it. This enables a localized and near-instantaneous sampling operation, which contrasts with other conventional sampling methods such as BAL, which introduce large amounts of fluid into the lungs and then remove them by suction, thus hindering breathing during the sampling operation by wedging the endoscope in place.

[0081] The distal end of the sheath may be adjacent to and / or substantially flush with the distal end of the first lumen or the first conduit and / or the distal end of the second lumen or the second conduit.

[0082] Alternatively, the distal ends of the first lumen or first catheter and / or the distal ends of the second lumen or second catheter (typically the distal ends of the first lumen or first catheter and the distal ends of the second lumen or second catheter) may extend beyond (i.e., in a more distal direction) the distal end of the sheath. This can facilitate the formation of a constant or stable volume of the first fluid and / or the second fluid in use by minimizing possible interactions and / or capillary action with the distal end of the sheath.

[0083] Advantageously, the space between the first and second lumens and the sheath can be filled or sealed with a filler material, at least at or near the distal end of the sheath. The filler material may include or may be an adhesive, such as epoxy resin. This arrangement prevents the first and / or second fluids from being drawn into the sheath, for example by surface tension or capillary action, which can help promote stable flow of the first and / or second fluids between the first or dispensing lumen and the second or suction lumen. This may be particularly advantageous when the distal end of the sheath is substantially flush with the distal end of the first lumen or the first conduit and / or the distal end of the second lumen or the second conduit.

[0084] It should be understood that, as long as the device achieves continuous or self-sustaining fluid flow between the first (dispensing) lumen and the second (suction) lumen during use, various configurations between the lumen and the probe are conceivable.

[0085] The first lumen and / or the first conduit may have a first width or diameter, such as a first internal width or diameter.

[0086] The second lumen and / or the second conduit may have a second width or diameter, such as a second internal width or diameter.

[0087] The second width or diameter (e.g., the second internal width or diameter) may be equal to or greater than the first width or diameter (e.g., the first internal width or diameter). Typically, the second width or diameter (e.g., the second internal width or diameter) may be greater than the first width or diameter (e.g., the first internal width or diameter). Advantageously, with such an arrangement, the first fluid may preferentially migrate or flow toward or into the second or suction lumen or second conduit. Advantageously, in the event of failure of the associated pumping mechanism (e.g., a pump), this may preferentially allow the first fluid to flow into the suction lumen.

[0088] Also advantageously, this allows the pressure of the fluid in the first or dispensing lumen or first conduit to be higher than the pressure of the first fluid in the second or aspiration lumen or second conduit. This can help provide a fluid barrier to prevent physical blockage of the aspiration lumen, and / or can help prevent blockage.

[0089] The first width or diameter (e.g., the first internal width or diameter) of the first lumen or the first conduit can be about 50-1000 μm, for example about 100-500 μm, for example about 100-200 μm, for example about 150 μm.

[0090] The second width or diameter (e.g., second internal width or diameter) of the second lumen or the second conduit can be about 50-1000 μm, for example about 200-800 μm, for example about 500-700 μm, for example about 580 μm.

[0091] Advantageously, the small size of the first and second catheters allows for the use of a very small total volume of sampling fluid (typically in the range of about 300 μL to 1.5 mL) to complete the sampling event. Typically, the minimum volume of sampling fluid required to perform the sampling event is greater than the "dead volume," i.e., the volume of the first catheter of the probe, which is typically about 300 μl. Therefore, the first fluid volume required to perform the sampling event can be greater than about 300 μl, for example greater than about 350 μl, and / or can be about 300 μl to 1.5 ml, for example about 350 μl to 1.5 ml. It should be understood that the precise minimum sampling fluid volume will depend on the precise dimensions of the probe, including the first lumen diameter and / or probe length. These small sample volumes can help minimize the dilution of the collected sample. Advantageously, these dimensions also enable fluid flow to generate shear rates similar to those experienced under normal physiological conditions to protect tissue and sample, the range of which is similar to the range of shear rates associated with commercially available syringe drivers, thereby preventing damage to the sampled tissue and / or the collected sample.

[0092] The ratio of the second width or diameter (e.g., the second inner width or diameter) to the first width or diameter (e.g., the first inner width or diameter) can be from about 5:1 to 0.8:1, for example from about 5:1 to 1:1, for example from about 4:1 to 2:1. The ratio of the second width or diameter (e.g., the second inner width or diameter) to the first width or diameter can be greater than 1:1.

[0093] It should be understood that the specific size of the first lumen and / or the second lumen may depend on many factors, including the type of the target being sampled (e.g., tissue or cells), and the above-mentioned sizes are provided only as background information, particularly for sampling ELF from the lungs of a subject.

[0094] The sampling device may also include a circulation device or a pumping device, such as one or more pumps.

[0095] The sampling device may include a first pump configured to dispense the first fluid via the first lumen.

[0096] The first pump can be configured to operate at a first flow rate.

[0097] The sampling device may include a second pump configured to draw the fluid, such as a first fluid, through the second lumen.

[0098] The second pump can be configured to operate at a second flow rate.

[0099] The first pump and the second pump can be the same.

[0100] A single pump may be present, configured to dispense the first fluid via the first lumen and draw the fluid via the second lumen. In such a case, the first flow rate and the second flow rate may be the same.

[0101] Advantageously, using a single pump can reduce the cost, size, and / or complexity of the device.

[0102] The first pump and the second pump can be different or distinct. Therefore, a first pump configured to dispense the first fluid via the first lumen and a second pump configured to draw the second fluid via the second lumen can be provided. In such a case, the first flow rate and the second flow rate can be the same or different.

[0103] Advantageously, using two independent pumps allows for adjustment of the relative value between the first and second flow rates. This can facilitate the regulation of the device's performance during use.

[0104] The circulation device or pumping device (e.g., one or more pumps) may form or define a sealed circuit or system. For example, the circulation device or pumping device (e.g., one or more pumps) may include an airtight injector.

[0105] Typically, the first flow rate and / or the second flow rate can be in the range of about 50-7000 μl / min, for example about 100-2000 μl / min, for example about 500 μl / min.

[0106] Advantageously, the first and second flow rates can be relatively low. This setup minimizes the shear rate, thereby avoiding or reducing damage to the cellular material in the fluid (e.g., the first fluid). Typically, the first and second flow rates can be less than about 5000 μl / min, for example, less than about 1000 μl / min.

[0107] Typically, when used in conjunction with lung sampling, the shear rate in the first lumen can be maintained at approximately 1700 Dyn / cm. 2 Below, for example, it remains at approximately 1000 Dyn / cm 2 the following.

[0108] Typically, when used in conjunction with lung sampling, the shear rate in the second lumen can be maintained at approximately 20 Dyn / cm. 2 Below, for example, it remains at approximately 10 Dyn / cm 2 the following.

[0109] It should be understood that the specific flow rate parameters used may depend on many factors, including the type of target being sampled (e.g., tissue or cells), and the parameters mentioned above are provided only as background information, particularly for ELF sampling from the lungs of a subject.

[0110] The sampling device can be used in conjunction with an endoscope (e.g., a bronchoscope). The endoscope or bronchoscope may also include a camera. Typically, the endoscope or bronchoscope may include or use a fiber optic camera.

[0111] While the advantage of the device of the present invention is its suitability for use in conjunction with conventional endoscopes, it should be understood that the device can be used without an endoscope. For example, the sampling device (e.g., its probe) can be deployed directly (e.g., in a “blind” manner) to the sampling location.

[0112] The first fluid may include or may be a liquid, such as a biocompatible liquid, like a salt solution, preferably a sterile salt solution, which can help minimize diffusion to achieve accurate local sampling.

[0113] The sampling device can be configured to sample a target area (e.g., target tissue) of the subject. In one embodiment, the sampling device can be configured to sample mucosal lining tissue (e.g., epithelial lining fluid (ELF)). In such a case, the second fluid may include or may be a mixture of the first fluid and ELF.

[0114] The endoscope or bronchoscope may also include an illumination device, such as one or more lamps.

[0115] The endoscope or bronchoscope may include a housing (typically a tubular housing). The endoscope or bronchoscope may include or define a port (e.g., a biopsy port) and / or an internal catheter through which the sampling device, such as a probe, can be inserted and / or deployed. Typically, the port and / or internal catheter may have a dimension of approximately 2 mm (e.g., internal width or diameter).

[0116] One or more components of the endoscope or bronchoscope (e.g., components of the endoscope or bronchoscope) may be housed within the housing.

[0117] As described above, the width (e.g., outer width or outer diameter) of the sheath of the device (e.g., probe) can be approximately 1-1.9 mm, for example 1-1.8 mm, or 1-1.7 mm, to allow the probe to be deployed via the endoscope cavity or lumen / endoscope cavity or lumen. This allows the device (e.g., probe) to be inserted into a conventional endoscope to deploy the distal end of the device to the sampling site. Advantageously, this arrangement prevents the distal end of the device from becoming contaminated, for example, due to contact with the subject's tissue surface. This ensures that the distal end of the device (e.g., probe) remains sterile and / or uncontaminated until it is deployed to the sample site via the distal end of the endoscope.

[0118] The endoscope or bronchoscope may include a directional / steering device and / or a directional / steering mechanism configured to adjust the orientation of the endoscope or bronchoscope (e.g., orientation at its distal end). The endoscope or bronchoscope may include Bowden cable, etc.

[0119] According to a second aspect, a system is provided, the system comprising:

[0120] Endoscope; and

[0121] Sampling device, wherein the sampling device includes a probe, wherein the probe includes:

[0122] A first lumen, the first lumen being configured to dispense a first fluid at its distal end; and

[0123] A second lumen, configured to aspirate the first fluid at its distal end, wherein the distal end of the first lumen is adjacent to the distal end of the second lumen.

[0124] The sampling device is configured to dispense the first fluid via the first lumen and simultaneously aspirate the first fluid via the second lumen.

[0125] The sampling device can be the sampling device according to the first aspect.

[0126] Advantageously, the probe can be configured to be inserted or deployed via a port or internal conduit of the endoscope. Advantageously, the external width or diameter of the probe can be smaller than the internal width or diameter of the port or internal conduit of the endoscope.

[0127] The endoscope may include or may be a bronchoscope.

[0128] The first fluid may include or may be a liquid, such as a biocompatible liquid, like a salt solution, preferably a sterile salt solution.

[0129] The sampling device can be configured to sample a target area (e.g., target tissue) of the subject. In one embodiment, the sampling device can be configured to sample epithelial liner fluid (ELF). In such a case, the second fluid may include or may be a mixture of the first fluid and ELF.

[0130] The endoscope (e.g., a bronchoscope) may also include a camera. Typically, the bronchoscope may include or may use a fiber optic camera.

[0131] The endoscope (e.g., a bronchoscope) may also include an illumination device, such as one or more lamps.

[0132] The endoscope (e.g., a bronchoscope) may include a housing (typically a tubular housing).

[0133] One or more components of the endoscope (e.g., a bronchoscope) may be housed within the housing. The probe and / or the camera (preferably, the probe and the camera) may also be housed within the housing.

[0134] The endoscope (e.g., a bronchoscope) can be a flexible endoscope, such as a bronchoscope.

[0135] The endoscope (e.g., a bronchoscope) may include a directional / steering device and / or a directional / steering mechanism configured to adjust the orientation of the endoscope (e.g., a bronchoscope) (e.g., orientation at its distal end). The endoscope (e.g., a bronchoscope) may include Bowden cables, etc.

[0136] The sampling device may also include a pumping device, such as one or more pumps.

[0137] The sampling device may include a first pump configured to dispense the first fluid via the first lumen.

[0138] The first pump can be configured to operate at a first flow rate.

[0139] The sampling device may include a second pump configured to draw the second fluid via the second lumen.

[0140] The second pump can be configured to operate at a second flow rate.

[0141] The features described with respect to the first aspect of the invention are equally applicable to the second aspect, and will not be repeated for the sake of brevity.

[0142] According to a third aspect, a method for sampling a target sample, such as a biological tissue, is provided, the method comprising:

[0143] A sampling device including a probe is provided, wherein the probe includes a first lumen and a second lumen, the first lumen being configured to dispense a first fluid at its distal end, and the second lumen being configured to aspirate the first fluid at its distal end, wherein the distal end of the first lumen is adjacent to the distal end of the second lumen.

[0144] Dispensing the first fluid via the first lumen; and

[0145] Simultaneously, the first fluid is drawn in through the second lumen.

[0146] The method may include establishing a self-sustaining and / or continuous fluid flow at its distal end between the first (dispensing) lumen and the second (suction) lumen.

[0147] Advantageously, the flow of the first fluid from the first lumen can help prevent the second lumen from becoming blocked during suction.

[0148] Advantageously, this method enables continuous sampling, for example by deploying the sampling device via an endoscope (e.g., a bronchoscope). Alternatively, the method can achieve continuous sampling by directly using the sampling device (e.g., its probe), for example by deploying the sampling device "blindly".

[0149] Typically, the method may include deploying the device, for example, placing the device (e.g., the distal end of the probe) at or near the target sample before circulating the first fluid.

[0150] The method may include deploying the sampling device "blindly" to the sampling location.

[0151] Alternatively, the method may include inserting the device (e.g., a probe) into the lumen or internal catheter of an endoscope to guide the probe to a target site and / or vicinity of a target sample. With this setup, the probe can be protected within the lumen or internal catheter of the endoscope until reaching the target site, thereby maintaining sample sterility and preventing contamination and confusion with other tissues / pathologies, thus providing true local sampling.

[0152] The method may include positioning the distal end of the device (e.g., a probe) in contact with or near the target sample (e.g., biological tissue). The term "near" will be understood herein to mean that, during use, such as during dispensing the first fluid and / or aspirating the first or second fluid, a fluid interface between the distal end of the probe and the target can be maintained. The method may include bringing the first fluid into contact with the target.

[0153] The method may include establishing a fluid interface between the first fluid and the target sample, and / or between the fluid flowing between the first (dispensing) lumen and the second (suction) lumen and the target sample.

[0154] The method may include placing the distal end of the device (e.g., a probe) at a distance of less than 2 mm, for example less than 1 mm, or for example less than 0.5 mm from the target. The method may also include bringing the distal end of the device (e.g., a probe) into contact with the target.

[0155] Advantageously, this method can achieve brief and / or gentle contact between the probe and the target in order to effectively position the probe while minimizing or avoiding damage to the target (e.g., biological tissue).

[0156] The method may include positioning the distal end of the device (e.g., a probe) close to or in contact with the target (tissue). For example, the method may include positioning the distal end of the device (e.g., a probe) at an angle (e.g., less than 90°, e.g., less than 80°) relative to the target (tissue). Advantageously, this can create at least a local gap or space between the distal end of the probe and the target, which allows the fluid interface to be maintained during use, such as during dispensing the first fluid and / or aspirating the second fluid, while preventing the probe (e.g., the first lumen or the second lumen) from becoming blocked when in contact with the target (tissue).

[0157] The method may include dispensing the first fluid continuously or semi-continuously at its distal end via the first lumen, and / or aspirating the first fluid or the second fluid at its distal end via the second lumen.

[0158] The method may include dispensing a fluid, such as a first fluid, with a total volume ranging from about 300 μL to 1.5 mL. Typically, the method may include dispensing a minimum volume of sampling fluid required to perform the sampling event, said minimum volume being greater than the “dead volume,” i.e., the volume of the first conduit of the probe, which is typically about 300 μl. Therefore, the method may include dispensing a first fluid volume greater than about 300 μl (e.g., greater than about 350 μl) and / or about 300 μl to 1.5 ml (e.g., about 350 μl to 1.5 ml). It should be understood that the precise minimum volume of the sampling fluid will depend on the precise dimensions of the probe, including the first lumen diameter and / or probe length. These smaller sample volumes may help to minimize the dilution of the collected sample. Advantageously, these dimensions also enable fluid flow to generate shear rates similar to those experienced under normal physiological conditions to protect tissue and sample, said shear rates being similar to the range of shear rates associated with commercially available syringe drivers, thereby preventing damage to the sampled tissue and / or the collected sample.

[0159] The method may include starting one or more pumps. Preferably, the one or more pumps are located within a sealed fluid circuit.

[0160] The method may include activating a first pump configured to dispense the first fluid via the first lumen. The first pump may be configured to operate at a first flow rate.

[0161] The method may include activating a second pump configured to draw the first fluid or the second fluid via the second lumen. The second pump may be configured to operate at a second flow rate.

[0162] The first pump and the second pump can be the same.

[0163] A single pump can be provided, configured to dispense the first fluid via the first lumen and draw the second fluid via the second lumen. In this case, the first flow rate and the second flow rate can be the same. Advantageously, using a single pump can reduce the cost, size, and / or complexity of the device.

[0164] The first pump and the second pump can be different or distinct. Therefore, the method can include activating a first pump configured to dispense the first fluid via the first lumen, and activating a second pump configured to draw the second fluid via the second lumen. In such a case, the first flow rate and the second flow rate can be the same or different. Advantageously, using two independent pumps allows for adjustment of the relative values ​​between the first and second flow rates. This can facilitate the tuning of the device's performance during use.

[0165] The method may include creating a fluid interface, such as a permanent and / or stable fluid interface, between the distal end of the probe and the target sample.

[0166] The method may include moving (e.g., translating) the device (e.g., a probe). Advantageously, this allows for sampling of a larger area of ​​the target sample while maintaining the fluid interface between the distal end of the probe and the target sample.

[0167] The first fluid may include or may be a liquid, such as a biocompatible liquid, like a salt solution, preferably a sterile salt solution.

[0168] The sampling device can be configured to sample a target area (e.g., target tissue) of the subject. In one embodiment, the sampling device can be configured to sample mucosal lining tissue (e.g., epithelial lining fluid (ELF)). In such a case, the second fluid may include or may be a mixture of the first fluid and ELF.

[0169] The method may include collecting fluid aspirated via the second lumen, such as a first fluid or a second fluid.

[0170] The method may include aspirating fluid (e.g., a first fluid or a second fluid) aspirated via the second lumen.

[0171] Features described with respect to any aspect of the invention are equally applicable to any other aspect, and will not be repeated for the sake of brevity only. For example, features described with respect to a composition may be applied to a method, and vice versa. Attached Figure Description

[0172] Embodiments of the present invention are described with reference to the accompanying drawings, wherein:

[0173] Figure 1 The system according to the first embodiment is shown;

[0174] Figure 2 An embodiment of the distal end of the probe of a sampling device according to another embodiment is shown;

[0175] Figure 3a Example 3c illustrates the distal end of the probe of a sampling device according to another embodiment;

[0176] Figure 4 The results of analyte measurements in fluids sampled in vivo using the conventional BAL method are shown.

[0177] Figure 5 Showing in with Figure 4 The same animal body but using Figure 1 The sampling equipment measures the analytes in the fluid sampled;

[0178] Figure 6 The illustrated chart shows the use of BAL with Figure 1 The sampling equipment samples the urea level in the fluid;

[0179] Figure 7 The illustrated chart shows the use of BAL with Figure 1 The sampling equipment samples the GM-CSF level in the fluid;

[0180] Figure 8 The chart shown is based on the total sample volume example illustration. Figure 1 The sampling equipment samples the GM-CSF concentration in the fluid;

[0181] Figure 9 Showing the use Figure 1 Microscopic images of epithelial lining fluid sampled by the device;

[0182] Figure 10 This shows the experimental timeline related to studies of potential inflammatory responses;

[0183] Figure 11 Showing with Figure 10 Standard curves related to potential inflammatory responses. Detailed Implementation

[0184] Throughout this disclosure, numerous terms are used, and unless the context otherwise requires, these terms have the meanings provided below. The nomenclature used herein to define compounds (particularly those according to the invention) is generally based on the nomenclature rules for chemical compounds established by the International Union of Pure and Applied Chemistry (IUPAC), specifically the "IUPAC Compendium of Chemical Terminology (Gold Book)". For the avoidance of ambiguity, if the IUPAC rules conflict with the definitions provided herein, the definitions herein shall prevail. Furthermore, if a compound structure conflicts with the name provided for that structure, the name provided for that structure shall prevail.

[0185] The term “comprising” or variations thereof should be understood herein to imply inclusion of the stated elements, integers or steps, or groups of elements, integers or steps, but does not exclude any other elements, integers or steps, or groups of elements, integers or steps.

[0186] The term “composition” or variations thereof should be understood to imply inclusion of the stated elements, integers or steps, or groups of elements, integers or steps, but does not exclude any other elements, integers or steps, or groups of elements, integers or steps.

[0187] When modifying numbers or values, the term "about" is used in this document to refer to values ​​within ±5% of the specified value. For example, if a temperature is specified as about 5 to about 13°C, this includes temperatures from 4.75 to 13.65°C.

[0188] References to the physical state of a substance (such as liquid or solid) refer to the state of the substance at 25°C and atmospheric pressure, unless the context otherwise requires.

[0189] Figure 1 A system 100 according to a first embodiment is shown.

[0190] System 100 includes a sampling device 115, and in this embodiment, includes an endoscope 110. The sampling device 115 includes a probe 120 and pumps 151 and 161. Reference is made below. Figure 2 -3 describes probe 120 in more detail; similar components are indicated by similar reference numerals, but with the value increased to "100". In this embodiment, endoscope 110 has a housing 111 and includes an internal catheter (not shown) through which probe 120 can be inserted and deployed. In this embodiment, sampling device 115 is configured to sample epithelial lining fluid (ELF) from the patient's lung 140 at the distal end 130 of probe 120.

[0191] Figure 2 Figures 1 and 3 show the distal ends 130 and 230 of probes 220 and 320, which can be coupled to, for example... Figure 1 The mirror bodies shown are used in combination.

[0192] like Figure 2As best shown, probe 220 includes a first lumen 222 and a second lumen 223, the first lumen being configured to dispense a first fluid at a distal end 230 of the probe, and the second lumen being configured to aspirate the first fluid at the distal end 230 of the probe. As shown, the distal end of the first lumen 222 is adjacent to the distal end of the second lumen 223. Advantageously, the space 224 between the first lumen 222 and the second lumen 223 and the sheath 221 is filled or sealed at least at or near the distal end of the sheath with a filler material. This prevents the sampling fluid from being drawn into the sheath 221, for example, by surface tension or capillary action. This can help facilitate a stable flow of the sampling fluid between the first lumen 222, or dispensing lumen, and the second lumen 223, or aspiration lumen.

[0193] The sampling device 100 is configured to dispense a first fluid via first lumens 222 and 322, and simultaneously draw in the first fluid via second lumens 223 and 323.

[0194] Advantageously, this enables continuous sampling, for example, when compared with... Figure 1 When used in conjunction with the endoscope 110 in system 100.

[0195] This is illustrated in Figures 3(a)-3(c). As shown in Figure 3(a), a first fluid 325 is dispensed at the distal end 330 of the probe 320 via a first lumen 322. As shown in Figure 3(b), suction via an adjacent second lumen 323 causes the first fluid 325 to be drawn into the second lumen 323 at the distal end 330 of the probe 320.

[0196] Advantageously, by controlling one or more parameters of the sampling device 100, such as the flow rate and / or size of the first lumen 322 and the second lumen 323, a self-sustaining flow of fluid 325 can be generated at the distal end 330 of the probe 320 between the first (dispensing) lumen 322 and the second (aspiration) lumen 323. This allows for simultaneous and / or continuous sampling in soft contact (fluid contact) with the target, thereby minimizing potential damage to the subject's tissues. Also advantageously, this allows the probe 120 to translate during sampling, thereby enabling normal function of the target (tissue), such as respiration.

[0197] Advantageously, the device can be configured to generate a substantially constant or stable volume of a first fluid 325 at or near the distal end 330 of the probe 320 during use. In this embodiment, the constant or stable volume of the first fluid 325 may define or may form droplets 325 (illustrated for ease of understanding). With the sheath 321 having an outer diameter of about 1.8 mm, the volume of the first fluid and / or droplets 325 may be about 0-200 μl, for example about 1-200 μl, for example about 10-200 μl.

[0198] This can help to create a continuous fluid interface between the probe 120 and the target (e.g., the subject's tissue) during use. Therefore, although the device 100 is able to generate a self-sustaining volume of fluid 325 at the distal end of the probe 320, it should be understood that when the fluid droplet comes into contact with the target / the target (e.g., tissue), surface tension will cause the fluid to "wet" the target (tissue), thereby creating a fluid interface, preferably a continuous fluid interface.

[0199] For clarity and ease of understanding, Figures 3(a)-3(c) illustrate probe 320 separately, so that the fluid drawn into the second lumen 323 is the same fluid as the fluid distributed via the first lumen 322.

[0200] However, as shown in Figure 3(d), it should be understood that in use, when the device is deployed at or near the target 341, and / or when a fluid interface is established with the target, the first fluid dispensed from the first lumen 322 “wets” the target and / or interacts with the target, thereby generating a second fluid 326. In such a case, the second lumen 323 is configured to draw in the second fluid 326 at its distal end 330. In other words, after a fluid interface is established with the target 341, the fluid 326 drawn in by the second lumen 323 is the second fluid 326 generated via the interaction between the first fluid 325 and the target sample 341.

[0201] like Figure 2 As shown in Figure 3, the probe includes sheaths 221 and 321. Typically, the first lumen 222 and 322 and the second lumen 223 and 323 are disposed within the sheaths 221 and 231.

[0202] The sheath has a distal end 235 at or near the sampling end 130 of the device 100. The width of the sheath (e.g., internal diameter) is typically about 0.5-5 mm, for example about 1-3 mm, such as about 1.8 mm. Suitably, this can be similar to or smaller than the internal diameter of the biopsy port of a standard bronchoscope (typically about 2 mm), which allows the probe 120 of the present invention to be placed within or compatible with a conventional bronchoscope 110.

[0203] exist Figure 2 In this embodiment, the distal end 235 of the sheath 221 is adjacent to and substantially flush with the distal ends of the first lumen 222 and the second lumen 223. In this configuration, the space between the first lumen 222 and the second lumen 223 and the sheath 221 is preferably filled with a filler material (not shown) at least at or near the distal end 235 of the sheath 221. This arrangement prevents the first fluid 325 from being drawn into the sheath 221, for example, by surface tension, which can help promote a stable flow of the first fluid between the first lumen 222, or distribution lumen, and the second lumen 223, or suction lumen.

[0204] Alternatively, as shown in Figure 3, the distal ends of the first lumen 322 and the second lumen 323 extend beyond (i.e., in a more distal direction) the distal end 335 of the sheath 321. This can facilitate the formation of a constant or stable volume of the first fluid 325 in use by minimizing possible interactions and / or surface tension with the distal end 335 of the sheath 321.

[0205] like Figure 1 As shown, the sampling device 115 includes a circulation device or a pumping device, here in the form of pumps 151 and 161.

[0206] The first pump 151 is in fluid communication with the first lumen 222 via the first conduit 153 and is configured to dispense a first fluid from the first fluid reservoir 152. The first pump is configured to operate at a first flow rate (approximately 500 μl / min in this embodiment).

[0207] The second pump 161 is in fluid communication with the second lumen 223 via the second conduit 163 and is configured to draw the first fluid into the second fluid reservoir 162. The second pump is configured to operate at a second flow rate (approximately 500 μl / min in this embodiment) such that the generated shear force does not exceed the shear force experienced by the system under physiological conditions. Advantageously, the use of two independent pumps 151, 161 allows for adjustment of the relative values ​​between the first and second flow rates. This can facilitate the adjustment of the performance of device 115 during use.

[0208] Although this embodiment features two separate pumps 151 and 161 for dispensing and aspirating the first fluid, it should be understood that in other embodiments, a single pump may be provided, configured to both dispense the first fluid via the first conduit 153 and aspirate fluid via the second conduit 163. In such a case, the first and second flow rates are substantially the same. Advantageously, using a single pump can reduce the cost, size, and / or complexity of the equipment.

[0209] Typically, the first flow rate and / or the second flow rate can be in the range of about 50-7000 μl / min, for example, about 100-2000 μl / min, for example, about 500 μl / min.

[0210] Advantageously, the first and second flow rates can be relatively low. This setting minimizes the shear rate, thereby avoiding or reducing damage to the cellular material in the fluid (e.g., the first fluid). Typically, the first and second flow rates can be less than about 5000 μl / min, for example, less than about 1000 μl / min.

[0211] Typically, when used in conjunction with lung sampling, the shear rate in the first lumen can be maintained at approximately 1700 Dyn / cm. 2 Below, for example, it remains at approximately 1000 Dyn / cm 2 the following.

[0212] Typically, when used in conjunction with lung sampling, the shear rate in the second lumen can be maintained at approximately 20 Dyn / cm. 2 Below, for example, it remains at approximately 10 Dyn / cm 2 the following.

[0213] Those skilled in the art will understand that endoscope 110 may include any devices that are typically useful in endoscopes but are not shown here for clarity, or may be associated with them, such as a camera (e.g., a fiber optic camera), an illumination device (e.g., one or more lamps), and / or a directional device and / or directional mechanism (e.g., Bowden cable) configured to adjust the orientation of probe 220 at its distal end 230.

[0214] It should be understood that this embodiment is provided as an example only, and various modifications can be made to this embodiment without departing from the scope of the invention.

[0215] Example

[0216] Overview

[0217] We conducted a preliminary in vivo study to validate a novel lung sampling method (using... Figure 1 The device, which has achieved promising results in a large animal sheep model that simulates the lungs of an adult, has been used.

[0218] The hypothesis validated by this study is that the novel sampling probe should be able to detect clinically relevant levels of the target analyte (in this case, granulocyte / macrophage colony-stimulating factor (GM-CSF) protein in the sampled ELF), and that the dilution factor of the ELF should be comparable to that in the BAL.

[0219] Materials and methods

[0220] breeding

[0221] The animals were housed at the Large Animal Research Facility (LARIF) in the Easter Bush campus and cared for and maintained by the team at that facility. Anesthesia, blood sampling, and bronchoscopy were all performed by the LARIF team.

[0222] anaesthetization

[0223] Propofol was administered systemically first, followed by general anesthesia maintained by isoflurane administered by a LARIF anesthesiologist.

[0224] Gene therapy treatment

[0225] The right lung lobe was designated as the control side, and the left lung lobe as the treatment side. One week prior to treatment, the right anterior lobe (RA) underwent bacteriological analysis (BAL) to confirm lung health via a non-inflammatory state (differential cell count). Treatment involved delivery of mouse granulocyte / macrophage colony-stimulating factor (mGM-CSF) packaged in a lentiviral vector (vGM173) to the left lung. Based on previous experience with similar multi-site delivery in newborn piglets (which was also well-tolerated), a total of 1e10 TU was delivered to five lung zones via multi-site delivery (2e9 TU each delivered in 10 ml volumes to LC, LVD1, LVD2, LVD3, and LCD). Viral reflux to the right control lobe was observed during infusion, necessitating the use of a completely untreated blank control (naive control).

[0226] Two sheep were administered the GM-CSF infusion on day 0, with a third companion animal serving as an untreated blank control. Based on previous research, GM-CSF levels were expected to peak on day 7, at which point one animal was sacrificed to verify GM-CSF detection by the probe. Once GM-CSF was confirmed by enzyme-linked immunosorbent assay (ELISA), the second sheep was allowed to continue the long-term study. The untreated blank control companion animal was housed with this second sheep.

[0227] Sampling probe

[0228] Before use, the custom-designed 2.2-meter long, 1.8-mm outer diameter dual-lumen sampling probe was cleaned three times with 6 ml of Virkon and then rinsed three times with 6 ml of water (H2O). The dispensing lumen 222 was connected to the Luer lock of a 1-meter airtight glass syringe (Hamilton), pre-filled with PBST to minimize the possibility of air ingress into the system. The delivery lumen 222 was then backfilled with PBS to minimize the risk of airlock in the small-diameter lumen. The collection lumen 223 was connected to the Luer lock of a 1 ml empty airtight glass syringe (Hamilton), and both syringes were loaded into an injection pump (Harvard Instruments). During sampling, both pumps were operated at the same flow rate (0.5 mL / min in this embodiment).

[0229] Sampling time point

[0230] For ethical reasons (to minimize any unnecessary suffering / prolongation of the experiment), the first sheep was sampled and euthanized on day 7, and additional ex vivo sampling was performed. GM-CSF was confirmed with a 35% higher in vivo level, justifying the use of a second animal for sampling. GM-CSF levels in the probe and BAL samples were quantified and compared using ELISA (Quantikine ELISA mouse GM-CSF, R&D systems). After confirming the detection of GM-CSF in BAL, probe, and serum samples, the second sheep was sampled by probe on day 7. The animal was sampled in the same manner on days 7, 21, 28, 77, and 112 (a total of 16 weeks, spanning Christmas / New Year). The experimental completion point was determined when GM-CSF was no longer detectable in either the probe or BAL samples at week 16. The animal was euthanized, and tissues were collected for HCR RNA-FISH imaging and DNA analysis to determine whether GM-CSF function was silenced by the immune system or whether transduced cells were actively removed by an immune T-cell response.

[0231] Sampling operation

[0232] Prior to anesthesia, three blood samples were obtained for hematological testing, serum enzyme / acute phase protein testing, and serum urea level quantification. A novel probe was used to sample segments RC, LC, LVD1, LVD2, LVD3, and LCD. During sampling, probe 120 was introduced through the biopsy port of endoscope 110. Before advancing the sampling catheter, the target area of ​​the airway was visually identified and examined; care was taken not to obstruct respiratory movement in any way. During sampling, the fixed probe tip effectively translated back and forth within the same airway area under the influence of respiration. Fluid collection into the collection syringe could be observed. The aspiration and dispensing pumps were programmed to automatically stop upon reaching the target volume.

[0233] Sample processing

[0234] Obtain GM-CSF levels in samples using a commercially available kit via ELISA (QuantikineELISA mouse GM-CSF, R&D systems) according to the manufacturer's instructions. Perform colorimetric assessment using a microplate reader (Synergy HT, Biotek) with the manufacturer's instructions and software (Gen5, Biotek).

[0235] Urea levels in samples were measured using a commercially available kit (MAK006 Sigma-Aldrich, Merck) according to the manufacturer's instructions. Colorimetric assessment was performed using a microplate reader (Synergy, Biotek) according to the manufacturer's instructions and software (Gen5, Biotek). ELF dilution was estimated by comparing ELF urea levels with serum levels obtained from Easter Bush pathological analysis of blood samples acquired by the LARIF team immediately prior to probe sampling.

[0236] result

[0237] The sampling method was repeatable and had a high success rate, with no hardware failures observed. Of the 51 samples from 3 animals, only 2 failed (suspected airlock in the system), and the recovery rate was relatively stable, approximately 75-90% of the infused sample volume (three times the recovery rate of BAL). All samples were clean; in contrast, BAL samples obtained over 16 weeks showed blood on multiple occasions. No tissue damage or irritation was observed during examination one week after sampling.

[0238] Interplate controls for ELISA remain comparable over several weeks, allowing for comparisons between different batches.

[0239] Comparison of BAL with novel probes

[0240] Figure 4The results of GM-CSF measurements in fluid sampled using BAL are shown.

[0241] Figure 5 The measurement results of GM-CSF in fluid sampled using an embodiment of the present invention as described above are shown.

[0242] from Figure 4 and Figure 5 As can be seen, using the BAL method and the novel probe method, GM-CSF was detectable in ELF sampled by the probe up to 77 days after instillation. On day 112 (16 weeks), GM-CSF was not detected, and the animals were euthanized. Therefore, the method of this invention can accurately measure the target analyte, with performance comparable to that obtained using the current "gold standard," BAL.

[0243] Figure 6 The illustrated chart shows the use of BAL with Figure 1 The sampling device samples the urea level in the fluid. This indicates that the urea concentration measured using the device of the present invention is comparable to the urea level measured using BAL, confirming the suitability of the device and method of the present invention as a sampling device (in the context of bronchoscopy in this application).

[0244] Figure 7 The illustrated chart shows the use of BAL with Figure 1 The sampling equipment samples the GM-CSF level in the fluid. This yields results related to the above regarding... Figure 6 The same conclusion is emphasized.

[0245] Figure 8 The chart shown is based on the total sample volume example illustration. Figure 1 The sampling equipment was used to sample the GM-CSF concentration in the fluid. Experiments were conducted with a first sample (smaller total volume, obtained in a 40-second sampling time) and a second sample (larger volume, obtained in a 120-second sampling time). It can be seen that sample volume does not affect the analyte concentration in the sample. Smaller sample volumes do not concentrate the analyte in the fluid sample. Extending the sampling time simply results in averaging over a larger sampling area. In other words, longer sampling times yield larger sample volumes without diluting the sample.

[0246] Figure 9 Showing the use Figure 1 Microscopic images of the epithelial liner fluid sampled by the device. The ELF sample was centrifuged at 450g for 5 minutes. 1 ml of supernatant was collected for downstream ELISA analysis. The remaining supernatant was discarded, and the cell pellet was resuspended in 1 ml PBS.

[0247] Cells were counted using a hematology counter, and the concentration was adjusted to 5 × 10⁻⁶. 5 / ml. 100 ml of this solution has a concentration of 5 × 10⁻⁶. 5 A sample of / ml was added to a cell centrifuge and centrifuged at 600 rpm for 5 minutes. The slides were then stained with Giemsa stain according to the manufacturer's instructions, and the slides were imaged at 20x magnification to produce... Figure 1 The image shown.

[0248] Investigation of potential inflammatory responses

[0249] We believe that the present invention offers significant advantages over existing methods: providing a self-sustaining volume of fluid at the distal end of the sampling device enables simultaneous and / or continuous sampling with soft contact (fluid contact) with the target, thereby minimizing potential damage to the subject's tissues compared to adsorption methods.

[0250] Based on the fact that no tissue damage was visually observed in the sampled airways during the aforementioned experiments, we believe this is indeed the case. This study aims to verify this hypothesis by quantifying the immune system response.

[0251] method

[0252] We treated a single segment (LVD1) of an adult ewe with gene-therapy virus (8 ml VGM124 2.8 e9 TU GLUX). We then sampled every four days over a 16-day period.

[0253] Experimental timeline at Figure 10 The following abbreviations are shown in the figure:

[0254] RC: Right heart lobe

[0255] LC: Left Heart Lobe

[0256] LVD1: Left ventral diaphragmatic lobe 1

[0257] LVD2: Left ventral diaphragmatic lobe 2

[0258] LCD: Left caudal diaphragm

[0259] "ELF" sampling was performed using the probe of this invention. Each sample was obtained over a 2-minute period using 1 ml of PBS.

[0260] Perform "BAL" as follows: wed the endoscope in, instill 10 ml of PBS through the bronchoscope biopsy channel, and then perform aspiration.

[0261] It should be noted that, for ethical reasons, BAL cannot be performed more than once a week in the same region, so the sampling area of ​​BAL will change over time.

[0262] All samples were tested using ELISA to detect the original solution (neat).

[0263] The board layout is shown in Table 1 below.

[0264] Table 1:

[0265]

[0266] The results of the two independent reads are shown in Tables 2 and 3 below.

[0267] Table 2: Readings at 450 nm:

[0268]

[0269] Table 3: Readings at 450 nm: 2

[0270]

[0271] The obtained standard curve is in Figure 11 As shown in the image.

[0272] result

[0273] The measurement results and related calculations are summarized in Table 4 below:

[0274] Table 4:

[0275]

[0276] The conclusion is that IL-6 levels were not significantly higher than the blank background level, and none exceeded the minimum standard of the standard curve.

[0277] This aligns with the initial hypothesis: since no physical damage was observed in the sampling area, no increase in IL-6 was induced by sampling, even with high-frequency sampling. Therefore, this study demonstrates that the sampling procedure caused no tissue damage, either visually or in terms of immune response.

[0278] Research on flow patterns

[0279] As mentioned earlier, intermittent flow can be used to distribute and / or pump fluid in a "semi-continuous" manner. This intermittent flow can be stable or unstable, such as ramp flow, square wave flow, oscillating flow, or pulsating flow. This study explores the impact of flow type on probe performance.

[0280] We hypothesize that effective sampling can still be achieved by changing the distribution flow pattern while maintaining a constant sampling flow rate; that is, we do not need to operate solely in continuous flow mode. Furthermore, we believe that discontinuous mode may be advantageous because the flow rate variation may generate stronger mixing at the sampling site, allowing more analyte to be mixed into the aspirate.

[0281] method

[0282] We treated a single segment (LVD1) of an adult ewe with gene therapy virus (8 ml VGM124 2.8 e9 TU GLUX) and sampled every four days for 16 days (4 sampling days).

[0283] At time points 1-4, the RC and LCD were sampled using different flow rate configurations.

[0284] The sampling event is:

[0285] -Sampling point 1: RC and LCD, using continuous aspiration and continuous dispensing, with a flow rate of 0.5 ml / min for both flows;

[0286] - Sampling point 2: RC and LCD, using continuous suction and zigzag ramp dispensing, the flow rate first ramps up to the maximum value, then ramps down to the minimum value and repeats in a cycle - ramping up from 0.25 ml / min to 0.75 ml / min within 6 seconds, then ramping down from 0.75 ml / min to 0.25 ml / min, and so on ( / \ / \ / \).

[0287] -Sampling point 3: RC and LCD, using continuous suction and square wave dispensing, switching between low and high rates, repeating with a 50% duty cycle, so the duration of high and low rates is equal -0.25 ml / min for 3 seconds, then 0.75 ml / min for 3 seconds, and so on (_|-|_|-|_).

[0288] -Sampling point 4: RC and LCD, using continuous aspiration and pulsed dispensing, intermittently switching from low flow rate to maximum flow rate in a short time, 1.19 ml / min for 1 second, then 0.36 ml / min for 5 seconds (|-|___|-|___).

[0289] Note that the flow parameters are balanced to ensure that the total volume of fluid allocated is the same in every case.

[0290] We measured the amount of protein in each sample (using the Pierce Protein Quantification Kit, Thermoscientific 16158), and for dilution, we monitored the level of endogenous urea in each sample (using Abcam ab83362) and the concentration of the exogenous marker mannitol (added to the allocated PBS) (using the Sigma-Aldrich MAK514 kit).

[0291] result

[0292] The results are shown in Table 5:

[0293]

[0294] As shown in Table 5, this study demonstrates that, in principle, any flow pattern can be used, and sampling can still be performed efficiently.

Claims

1. A sampling device including a probe, wherein, The probe includes: A first lumen, the first lumen being configured to dispense a first fluid at its distal end; A second lumen, configured to aspirate the first fluid at its distal end, wherein the distal end of the first lumen is adjacent to the distal end of the second lumen. The sampling device is configured to dispense the first fluid via the first lumen and simultaneously aspirate the first fluid via the second lumen.

2. The sampling device according to claim 1, wherein, The device is configured to generate a self-sustaining and / or continuous fluid flow between the first lumen and the second lumen at its distal end.

3. The sampling device according to claim 1 or claim 2, wherein, The sampling device is configured to generate a first fluid volume at or near the distal end of the probe during use, the volume being sufficient to create a continuous fluid interface between the probe and the target.

4. The sampling device according to any of the preceding claims, wherein, In use, when the sampling device is deployed at or near the target, and / or when a fluid interface is established with the target, the first fluid interacts with the target to generate a second fluid containing the analyte of interest through mixing.

5. The sampling device according to claim 4, wherein, The second lumen is configured to aspirate the second fluid at its distal end.

6. The sampling device according to any of the preceding claims, wherein, The sampling device is configured to continuously dispense a first fluid at the distal end of the first lumen via the first lumen, and to continuously aspirate the first fluid or the second fluid at the distal end of the second lumen via the second lumen.

7. The sampling device according to any of the preceding claims, wherein, The probe includes a sheath, wherein the first lumen and the second lumen are disposed within the sheath.

8. The sampling device according to claim 7, wherein, The outer width or outer diameter of the sheath is about 1-1.9 mm, or optionally about 1-1.8 mm.

9. The sampling device according to any of the preceding claims, wherein, The distal end of the first lumen is substantially flush with or level with the distal end of the second lumen, and the space between the first lumen and the second lumen and the sheath is filled or sealed with a filler material at least at or near the distal end of the sheath.

10. The sampling device according to any of the preceding claims, wherein, The first cavity has a first internal width or internal diameter, and the second cavity has a second internal width or internal diameter, wherein the second internal width or internal diameter is equal to or greater than the first internal width or internal diameter.

11. The sampling device according to claim 10, wherein, The ratio of the second inner width or inner diameter to the first inner width or inner diameter is in the range of approximately 22:1 to 1:

1.

12. The sampling device according to any of the preceding claims, further comprising one or more pumps configured to dispense the first fluid and / or aspirate the first fluid or the second fluid.

13. The sampling device according to claim 12, wherein, The sampling device includes a first pump and a second pump, the first pump being configured to dispense the first fluid via the first lumen at a first flow rate, and the second pump being configured to draw fluid / the fluid via the second lumen at a second flow rate.

14. The sampling device according to claim 13, wherein, The first flow rate and / or the second flow rate are in the range of about 50-7000 μl / min, and optionally in the range of about 100-2000 μl / min.

15. The sampling device according to any of the preceding claims, wherein, One or more of the first lumens are defined by a first conduit, and one or more of the second lumens are defined by a second conduit, wherein the first conduit and the second conduit are arranged side by side at least at the distal end of the sheath.

16. A system comprising: Endoscope; as well as Sampling device, wherein the sampling device includes a probe, wherein the probe includes: A first lumen, the first lumen being configured to dispense a first fluid at its distal end; as well as A second lumen, configured to aspirate the first fluid at its distal end, wherein the distal end of the first lumen is adjacent to the distal end of the second lumen, wherein the sampling device is configured to dispense the first fluid via the first lumen and simultaneously aspirate the first fluid via the second lumen. The probe is configured to be inserted or deployed via a port or internal conduit of the endoscope, and the outer width or outer diameter of the probe is smaller than the inner width or inner diameter of the port or internal conduit of the endoscope.

17. The system according to claim 16, wherein, Endoscopes include or may be bronchoscopes.

18. A method for sampling a target sample, such as biological tissue, the method comprising: A sampling device including a probe is provided, wherein the probe includes a first lumen and a second lumen, the first lumen being configured to dispense a first fluid at its distal end, and the second lumen being configured to aspirate the first fluid at its distal end, wherein the distal end of the first lumen is adjacent to the distal end of the second lumen. The first fluid is distributed via the first lumen; as well as Simultaneously, the first fluid is drawn in through the second lumen.

19. The method of claim 18, further comprising placing the distal end of the probe close to the target sample to establish a fluid interface between the distal end of the probe and the target.

20. The method of claim 18 or claim 19, comprising continuously dispensing the first fluid at a distal end of the first lumen via the first lumen, and continuously aspirating the first fluid or the second fluid at a distal end of the second lumen via the second lumen.

21. The method of any one of claims 18 to 20, the method comprising activating a first pump and activating a second pump, the first pump being configured to dispense the first fluid via the first lumen at a first flow rate, and the second pump being configured to draw the first fluid or the second fluid via the second lumen at a second flow rate.

22. The method according to claim 21, wherein, The first pump and the second pump are identical.

23. The method according to claim 21, wherein, The first pump and the second pump are different.

24. The method according to any one of claims 18 to 23, the method comprising generating a fluid interface between the distal end of the probe and the target sample.

25. The method according to any one of claims 18 to 24, the method comprising translating the probe relative to the target sample.

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