Patent Application
20240001359
The present subject matter relates, in general, to in-vitro biosensing and analysis of biological samples, and particularly but not exclusively relates to a microfluidic analyser for in-vitro biosensing and analysis of a biological sample.
In-vitro biosensing, analysis, and diagnostics play an important role in medical decision-making process. An in-vitro biosensing, analysis, and diagnostics process includes performing bioassays of a biological sample, such as blood, saliva, etc., taken from a subject. Examples of bioassays includes, but are not limited to, electrochemical assays, nucleic acid tests, enzyme activity assays, cell-based assays, and immunoassays. In the bioassays, various pre-treatment process steps may be involved, in which various reagents and other pre-treatment solutions may be introduced in an assay for pre-treatment of the biological sample.
The features, aspects, and advantages of the subject matter will be better understood with regard to the following description and accompanying figures. The use of the same reference number in different figures indicates similar or identical features and components.
Generally, an in-vitro biosensing process of a biological sample has three stages including sample processing, sample enrichment, and sample detection. In all the three stages, the biological sample, typically a liquid sample, is manually handled using high precision liquid handling systems, such as pipettes. Such manual handling of samples by a user in different instances or by different users, may vary with a high degree, introducing undesired subjectivity to the biosensing and diagnostics processes. Further, to eliminate or reduce the degree of subjectivity, the user may be required to use the liquid handling systems precisely, which increases an overall time required for performing the biosensing and diagnostic processes. Thus, the conventional biosensing and diagnostics processes require extensive training of the user.
Moreover, handling of the biological sample is required to be done in a contained infrastructure, so as to reduce potential damages to a technician or a user involved. In addition, a detection technology being used has to be targeted towards a specific biomarker from the biological sample while preventing false positive and false negative outcomes. Therefore, the conventional techniques extensively require specialized infrastructure and high precision equipment, which, in turn, increases the overall cost of performing biosensing and in-vitro diagnostics and analysis.
In this respect, various automated devices have been developed to carry out the biosensing and in-vitro diagnostics and analysis without manual intervention. However, the conventional devices for automated biosensing and in-vitro diagnostics and analysis are optimized to operate with lesser resources, such as various equipment, at a low resource setting. The conventional devices, involving high resource settings require centralized laboratories and involves implementation of specialized and bulky equipment.
In addition, the overall time taken for biosensing and diagnostics play a critical role in diagnosing a medical condition and initiating an appropriate treatment process in order to impart optimal clinical outcomes in a timely manner. However, the conventional devices, as well as the analysis reporting processing time-consuming which introduces a critical challenge in achieving optimal clinical outcomes in a timely manner.
The present subject matter relates to a device for processing a biological sample for detection and analysis of a biomarker. Examples of the biomarker may include, but are not limited to, protein, nucleotides, metabolites, and carbohydrates/lipids, immunosensors, deoxyribonucleic acid (DNA) bio-sensors, enzyme-based bio-sensors, tissue-based bio-sensors, and thermal bio-sensors. The microfluidic analyser of the present subject matter can simultaneously process multiple samples for bioassay. For example, the microfluidic analyser may perform a bioassay including, but not limited to, basic enzyme-linked immunosorbent assay (ELISA), DNA detection.
The microfluidic analyser of the present subject matter includes a platform, a fluid control unit coupled to the platform, and an optical unit operably coupled to the platform. The platform is configured to hold at least one cartridge carrying the biological sample and at least one reagent, for simultaneously performing in-vitro diagnostic evaluation. In an example, the at least one cartridge includes one or more sealed ends.
Further, the fluid control unit is configured to regulate flow of the biological sample and the at least one reagent inside the at least one cartridge. The fluid control unit may include one or more needles to pierceably connect with the one or more sealed ends of the at least one cartridge to establish a fluid connection with the at least one cartridge. The fluid control unit may also include a pneumatic unit, operably coupled to the one or more needles, to provide at least one of a positive pressure and a negative pressure to the at least one cartridge. In addition, the optical unit comprises an optical sensor to detect presence of a fluorescence biomarker in the biological sample held in the at least one cartridge.
In an example, the microfluid analyser also includes a linear guide mechanism, a controller, and a battery. The linear guide mechanism may be positioned below the platform and may enable movement of the optical unit to align the optical unit with the at least one cartridge. For example, in an event of simultaneous processing of multiple biological samples, the linear guide mechanism facilitates the optical sensor to be aligned below a specific cartridge.
Further, the controller may control the pneumatic unit to perform pre-processing of the sample. The controller may include a communication module to connect the microfluidic analyser with a remotely located centralized server, such as a cloud server. The controller may gather the bioanalysis and diagnostics results from the optical unit and transmit the results to the remotely located centralized server for real-time decision-making process. The battery allows a portable use of the microfluidic analyser. Due to portability, the microfluidic analyser is suitable for being used in remote locations where there is scarcity of electricity.
Accordingly, the present subject matter describes a compact and deployable microfluidic analyser for automated in-vitro diagnostics for processing biological samples to derive test results without any manual intervention. The microfluidic analyser is capable of self-containment and reagent processing, and waste disposal, thereby eliminating usage of additional and specialized infrastructure. As the microfluidic analyser of the present subject matter is automated, the microfluidic analyser is usable with minimum training requirement.
The microfluidic analyser is further equipped with communication capabilities, utilizing which the microfluidic analyser can share the diagnostics results to a remote location, through a cloud server, for real-time and continuous data analysis. Therefore, the microfluidic analyser expedites the overall processing of the samples in order to achieve optimal clinical outcomes.
These and other advantages of the present subject matter would be described in a greater detail in conjunction with
The platform 102 is configured to hold at least one cartridge carrying the biological sample and at least one reagent. The at least one cartridge includes one or more sealed ends. The platform 102 may have a plate shaped structure. For example, the platform 102 may be designed to hold at least one cartridge and the platform 102 may allow for a simultaneous analysis of multiple samples contained in the at least one cartridge. The platform 102 may be divided into a set of sections suitable for holding the at least one cartridge. In an example, each section from the set of sections may include a set of slots formed corresponding to the optical sensor 124 to allow the light beam from the optical sensor 124 onto the sample contained in the at least one cartridge.
In an example, the set of sections may include a retaining member (not shown) for locking the cartridges in a specific section, once the cartridge is positioned on the platform 102. The locking of the cartridges by the retaining member prevents an undesired movement of the cartridges while performing a sample analysis process.
In an example, the platform 102 includes a plurality of temperature-controlled zones. The plurality of temperature-controlled zones may be formed for maintaining a desired temperature of the at least one cartridge for pre-treatment of the biological samples in order to prepare the samples for analysis. In an example, the platform 102 includes a heating element (not shown in
For example, the platform 102 may include a nichrome wire-based structure, as the heating element, for electrical temperature management. Further, a set of temperature sensors may be provided corresponding to the temperature-controlled zones for measuring temperature values of the respective zones. The nichrome wire-based structure and the set of temperature sensors may be communicatively coupled to the controller 116. The controller 116, upon receiving measured temperature values from one of the temperature sensors, may precisely adjust the temperature of a corresponding zone by regulating a power delivered to the nichrome wire-based structure.
In an example, the controller 116 includes a communication module (not shown). The communication module may facilitate in establishing a cloud-based connectivity of the microfluidic analyser 100, and thus allowing for cloud connectivity of data being collected by the microfluidic analyser 100 by analysing the biological sample. For example, the controller 116 may include an Internet of things (IoT) module for allowing a remote connection of the microfluidic analyser 100 with a centralized server. The capability of the microfluidic analyser 100 to remotely store the collected data allows for remote classification and distribution of the collected data while ensuring security of the collected data.
In an example, the microfluidic analyser 100 comprises a covering member (not shown in
Further, the linear guide mechanism 106 is arranged below the platform 102 to align the optical unit 104 with the at least one cartridge. The linear guide mechanism 106 may allow a linear movement of the optical unit 104 corresponding to the platform 102. The linear movement of the optical unit 104 with the linear guide mechanism 106 may allow for aligning the optical sensor 124 with respect to the corresponding slots of the platform 102 for performing the analysis of the sample contained in the at least one cartridge.
In an example, the linear guide mechanism 106 includes a drive and a movable member. The drive may actuate a linear movement of the movable member. The movable member may be coupled to the optical unit 104, and the optical unit 104 may be moved linearly in conjunction with the movement of the movable member. Examples of the movable member include, but are not limited to, a belt and pully arrangement, a profiled rail, and a rack and pinion arrangement.
Further, the fluid control unit 108 is configured to regulate flow of the biological sample and the at least one reagent inside the at least one cartridge. The one or more needles 110 of the fluid control unit are aligned with the platform 102 to be able to pierceably connect with the one or more sealed ends of the at least one cartridge. Such connection allows to establish a fluid connection of the one or more needles 110 with the at least one cartridge. Further, the pneumatic unit 112 is operably coupled to the one or more needles 110, to provide at least one of a positive pressure and a negative pressure to the at least one cartridge.
Upon placement of the cartridge on the platform 102, the pneumatic unit 112 may be coupled to the cartridge through the one or more needles 110. The pneumatic unit 112 may provide controlled air pressure to the cartridge. The said air pressure may allow control of a sample or a sample treatment solution present in the cartridge. For example, the air pressure provided by the pneumatic unit 112 may allow movement of the sample and a target sample treatment solution, within the cartridge, towards a target area. Further, the with controlled air pressure, a processed sample may be moved towards a waste containment area. Similarly, an undesired portion of the sample may be moved within the cartridge for isolation and collection.
In an example, the pneumatic unit 112 is configured to open or close an air passage to the cartridge in order to control an ambient pressure inside the cartridge.
The platform 102, linear guide mechanism 106, and the pneumatic unit 112 may be communicatively coupled with the controller 116. The controller 116 may provide control signals in order to precisely control a function of any of the platform 102, linear guide mechanism 106, and the pneumatic unit 112. Further, the controller 116 may be powered by the battery 118. Powering the microfluidic analyser 100 by the battery 118 may allow for a portable usage of the microfluidic analyser 100. In another example, the microfluidic analyser 100 may be powered by an external power source.
In an example, the microfluidic analyser 100 includes a plurality of buttons coupled to the controller 116 and the display unit 120 to display to a user a set of control parameters and status of the microfluidic analyser 100. In an example, the microfluidic analyser 100 may include a touch-sensing display unit 120 which may be used to control the control parameters of the microfluidic analyser 100.
In operation, a cartridge may be placed onto the platform 102 in a designated section of the platform 102. In the present operation described hereinafter, only one cartridge has been taken into account for the sake of brevity. However, the platform 102 may support placement of a plurality of cartridges and may support simultaneous analysis of a plurality of samples. Upon placement of the cartridge, the retaining member of the platform 102 a lock the cartridge in place. Further, the covering member may cover the cartridge from above and provide additional stability to the cartridge. The optical unit 104 is aligned, through the linear guide mechanism 106, with a section on the platform 102 containing the cartridge. Upon successful alignment of the optical unit 104 with the respective section, the controller 116 of the microfluidic analyser 100 may control the temperature of the plurality of temperature-controlled zones for pre-treatment of the sample.
The pneumatic unit 112 may be used to control an air pressure within the cartridge in order to perform pre-processing or pre-treatment of the sample with various reagents contained in the cartridge. The pre-processing of the sample may involve disintegrating a biochemical structure of the sample. Further, the pre-processing of the sample may involve mixing the sample with a washing reagent to remove undesired material from the sample.
In order to perform the above-described pre-processing step, upon placement of the cartridge, at least one outlet of the pneumatic unit 112 may couple with at least one opening of the cartridge. Upon successful coupling of the outlet of the pneumatic unit 112 with the inlet of the cartridge, the pneumatic unit 112 mat be automatically controlled by the controller to apply negative or positive pressure. Alternatively, the controller may also control the pneumatic unit 112 to open or close the inlet of the cartridge in order to control an internal pressure of the cartridge, without applying a negative of positive pressure. The operations of the pneumatic unit 112 may be performed by at least one solenoid valves. The said operations may result in the movement of the sample and a target reagent, from amongst the reagents, within the cartridge, allowing the performing of required pre-processing steps.
The optical sensor 124 may incident a light beam onto a sample and collect an emission from the sample generated due to the illumination by the incident light beam. The collection of the emission from the sample may involve detection of biosensors present in the sample. In an example, the biosensors are fluorescence markers, and the sample is marked with the fluorescence markers.
The display unit 120 may be communicatively coupled with the controller and may display a status of the microfluidic analyser 100 and control parameters associated with the microfluidic analyser 100. The display unit 120 may be coupled with a set of buttons for allowing a user to adjust and view different parameters of the microfluidic analyser 100.
Referring to
Referring now to
Referring now to
The one or more sealed ends 304 of the cartridge 300 may be couplable with a needle assembly of a fluid control unit, such as the fluid control unit 108. In an example, the cartridge 300 includes four sealed ends 304. The one or more sealed ends 304 may be air-tightly sealed in a non-operational state. In an operational state of air inlet, from amongst the one or more sealed ends 304, the air inlet may receive one of a positive pressure and a negative pressure from one of the control valves. Alternatively, opening and closing of the air inlet may be controlled through the control valve. The control of the pressure to the one or more sealed ends 304 and the respective opening and closing of the one or more sealed ends 304 may allow for a movement of the plurality of reagents, treatment solutions, and the sample within different chambers and regions of the cartridge 300.
The body 302 includes an opening 316 for receiving the biological sample. For example, the biological sample may be collected on a swab and the swab is inserted in the cartridge 300 through the opening 316. In an example, the received sample is collected in the storage chamber 306. In the storage chamber 306, the biological sample may be suitably pre-treated and prepared for further processing. In an example, the storage chamber 306 may be provided with a pre-stored solution that enables the pre-treatment of the sample. For example, the pre-stored solution is a buffer solution.
Further, the storage chamber 306 may be coupled to the processing chamber 308 through one of the plurality of channels 310. The processing chamber 308 may include a filtering member to filter the biological sample. In an example, the processing chamber 308 may include multiple filtering members. The processing chamber 308 may be coupled with a treatment media storage. In an example, the treatment media storage may be a serpentine flow channel. The treatment media storage may be pre-stored with a plurality of reagents and treatment solutions. The treatment solutions facilitate in selecting a target biomarker in the biological sample. For example, the treatment solutions may bind with an antibody present in the biological sample, thereby selecting the target biomarker.
Upon completion of pre-treatment of the biological sample, the biological sample may be directed to the detection region 312, by controlling pressure inputs to at least one of the one or more sealed ends 304. Upon reaching the detection region 312, an optical detector of the microfluidic analyser 100 of
The identification marker 314 may include a Quick Response (OR) code. The OR code may be readable by a OR code reader of an optical unit, such as the optical unit 104. The QR code may allow for an identification of the biological sample contained in the cartridge 300. The identification of the cartridge 300 may allow for proper indexing of the biological samples while preventing inter-mixing of analysis results of different samples.
The cartridge 300 may further include a waste collection chamber 318 to collect residual and processed reagents and the biological sample. The waste collection chamber 318 prevents other chambers to come in direct contact of the residual and processed reagents and sample. Therefore, the waste collection chamber 318 prevents potential contamination of contents of other chambers.
In an example, the cartridge 300 may be formed from plastic. For example, the cartridge 300 may be formed from one of a thermoplastic material, a polypropylene material, a polycarbonate material, a polymethylmethacrylate material, and a cyclic olefin copolymer material.
Although the cartridge 300 has been depicted to include a serpentine shaped channel carrying one or more reagents and a section for holding a buffer solution in which the biological sample is received, the cartridge may have varying configuration and design. Accordingly, the microfluidic analyser of the present subject matter may be configured to operate with cartridges of different sizes and designs.
In the implementation as depicted in
In the present implementation, a length of the platform 500 may be in a range of about 290 mm to about 300 mm. For example, the length of the platform 500 is 292.5 mm. Further, a width of the platform 500 may be in a range of about 85 mm to about 95 mm. For example, the width of the platform 500 is 91.22 mm. In addition, a width of each section of the platform 500 may be in a range of about 40 mm to about 50 mm. For example, the width of each section of the platform 500 is 45.1 mm. Also, a height of the platform 500 may be in a range of about 15 mm to about 25 mm. For example, the height of the platform 500 is 19.93 mm.
The first section 802 and the second section 804 when connected with each other, form an enclosure 806. The enclosure 806 may accommodate a fluorescent detector (not shown) and a Quick Response (QR) code detector (not shown). The fluorescent detector may allow for a detection of fluorescence biomarkers in a biological sample. The QR code detector may allow for an identification of a sample contained in a cartridge by reading a QR code which may be marked on the cartridge. The identification of the cartridge allows for preventing inter-mixing of analysis results of different samples.
Further, the enclosure 806 may be formed for precise alignment and assembly of above-mentioned optical components of the optical sensor 800. In an example, upon assembly of the optical components, an internal degree of freedom of the optical components may be restricted to enable long term detection without requirement for calibration. In an example, the optical sensor 800 may be coupled to a controller (not shown) of the microfluidic analyser. The controller may be optimized to minimize a dark current and enable high signal to noise ratio detection through the optical sensor 800.
The optical sensor 800 may be configured for quantification of DNA amplification of the sample contained in the cartridge using a custom optical detector.
As shown in
Further, as shown in
In operation, the optical sensor 800 may incident a light beam on a biological sample through the excitation filter. The incident beam, upon passing through the excitation filter may be incident on the biological sample. The optical sensor 800 may accordingly detect fluorescence emission caused by the illumination of the biological sample due to the incident light beam. Such emitted light beam from the biological sample may be passed through the emission filter. The dichroic mirror 810 may be provided to separate the excitation and emission light beams. The fluorescence detection may be used for performing a process of bioassay of the sample.
In an example, a length of the linear guide mechanism 900 may be in a range of about 280 mm to about 290 mm. For example, the length of the linear guide mechanism 900 is 288.7 mm. Further, a width of the linear guide mechanism 900 may be in a range of about 85 mm to about 95 mm. For example, the width of the linear guide mechanism 900 is 90.92 mm. In addition, a height of the linear guide mechanism 900 may be in a range of about 60 mm to about 70 mm. For example, the height of the linear guide mechanism 900 is 64.3 mm.
In an example, the fluid control unit 1000 includes one or more needles 1002 to pierceably connect with one or more sealed ends (not shown in
The fluid control unit 1000 further includes a plurality of tubes 1006 connected, at a first end 1006A, to a free end 1002B of the one or more needles 1002. In an example, the plurality of tubes 1006 are made of silicon. The fluid control unit 1000 also includes a plurality of control units 1008 which are coupled to a second end 1006B of the plurality of tubes 1006. The plurality of control units 1008 controls the flow of fluid from the pneumatic unit 1004 to the plurality of tubes 1006. For example, a control unit from the plurality of control units 1008 is coupled to an individual tube from the plurality of tubes 1006 to control the flow of fluid in the corresponding tube. In an example, the plurality of control units 1008 are electronically controlled valves, such as solenoid valves.
The fluid control unit 1000 further includes a plurality of check valves 1010. In an example, the plurality of check valves 1010 are mounted between the one or more needles 1002 and the plurality of control units 1008, to allow unidirectional flow of the fluid through the plurality of tubes 1006. In an example, a set of check valves 1010 may allow a flow of the fluid, through the plurality of tubes 1006, from the pneumatic unit 1004 towards the one or more needles 1002. Further, another set of check valves 1010 may allow a flow of the fluid, through the plurality of tubes 1006, from the one or more needles 1002 towards the pneumatic unit 1004. The unidirectional flow of the fluid controlled by the plurality of check valves 1010 may selectively provide a positive pressure or a negative pressure to the at least one cartridge.
The fluid control unit 1000 also includes a plurality of flow control valves 1012. In an example, the plurality of flow control valves 1012 are mounted between the one or more needles 1002 and the plurality of control units 1008. The plurality of flow control valves 1012 regulates the positive pressure or the negative pressure of the fluid provided at the at least one cartridge.
Further, the pneumatic unit 1004 may include a pump 1014, a check valve 1016, a reservoir 1018, a pressure sensor 1020. The reservoir 1018 may carry the fluid and the pump 1014 may be used to control the positive or negative pressure of the fluid in the reservoir 1018. The reservoir 1018 may include an inlet connected to the check valve 1016. The pump 1014 and the check valve 1016 are electronically controlled by a controller (not shown) of the microfluidic analyser to achieve a desired pressure value from the reservoir 1018. Further, the pressure sensor 1020 is coupled to the reservoir 1018 to measure a value of pressure of the reservoir 1018.
In an example, the needle assembly 1100 may include four set of inlet openings and outlet openings. For example, each set of inlet openings and outlet openings includes five needles. The needle assembly 1100 may equally distribute the incoming pressure from the valves to the four outlet openings.
In an example, the plurality of control units 1204 includes four or more number of valves having dedicated functions with respect to the controlling of the air pressure inside the cartridge. The valves may be configured to perform different operations, such as providing a positive pressure by addition of air in the cartridge, providing a negative pressure by removal of air from the cartridge, and opening and closing of an air passage of the cartridge.
By controlling a combination of the above-described configurations of the valves, a target liquid inside the cartridge can be moved to a specific desired direction or position.
The pump 1208 may control the positive or negative pressure in the pressure reservoir 1206. The pressure reservoir 1206 may include a set of inlets connected to the valves. The pump 1208 and the valves may be electronically controlled by a controller of the microfluidic analyser for achieving desired automation of liquid handling.
In an example, a length of the pneumatic unit 1202 may be in a range of about 135 mm to about 145 mm. For example, the length of the pneumatic unit 1202 is 141.55 mm. Further, a width of the pneumatic unit 1202 may be in a range of about 95 mm to about 100 mm. For example, the width of the pneumatic unit 1202 is 96.7 mm. In addition, a height of the pneumatic unit 1202 may be in a range of about 55 mm to about 65 mm. For example, the height of the pneumatic unit 1202 is 57.03 mm.
Although examples for the present disclosure have been described in language specific to structural features and/or methods, it is to be understood that the appended claims are not limited to the specific features or methods described herein. Rather, the specific features and methods are disclosed and explained as examples of the present disclosure.
| Number | Date | Country | Kind |
|---|---|---|---|
| 202011051838 | Nov 2020 | IN | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/IN2021/051106 | 11/26/2021 | WO |
