Patent Application
20210291179
The field of the invention is small volume mixers for research materials and pharmaceuticals.
Microfluidic mixing incorporates the physics of fluids flowing in small channels to promote self-assembly of nanoparticles that can encapsulate nucleic acids, small molecules, proteins and/or peptides efficiently and with minimal loss of the delicate and expensive materials. The resulting formulations are useful for academic research and medical treatment.
United States Publication Nos. 20120276209 and 20140328759, by Cullis et al. describe methods of using small volume mixing technology and novel formulations derived thereby. United States Publication No. 20160022580 by Ramsay et al. describes more recent advances using small volume mixing technology and products.
In recent years, devices for biological microfluidic mixing have been designed. Precision NanoSystems Inc., in Vancouver, Canada, manufactures and distributes such devices under the NanoAssemblr™ brand. Single use cartridges or microfluidic chips (hereinafter, “m-Chips”) are miniaturized, laboratory-ready mixing platforms which work within these devices.
Currently, control of the mixing process on an m-chip is exerted by an operator through machine controls or a manually operated mechanism. Under the direction of the operating technician or “user”, these dispense reagents to the inlet in the M-chip at an optimal speed in order to achieve optimal mixing.
The fluidic elements being mixed by researchers are increasingly complex and valuable, and include nucleic acids, peptides, and small molecule drugs. In the laboratory and in the case of personalized medicine, a greater understanding of which lipid/surfactant/drug ratios and particle size are optimal for each drug and tissue target require that large numbers of formulations be prepared and screened, each with specific conditions which must be carefully tracked. Furthermore, the M-chips are so small that a user cannot easily determine whether they are clean or soiled, free flowing or blocked.
There is a need for a semi-automated, quality-controlled microfluidic mixing apparatus, which reduces losses of expensive materials to an absolute minimum, and which enables consistently high quality formulations regardless of the experience of the user.
According to embodiments of the inventions, there is provided an instrument for mixing, said instrument having a motor, a pump, a microfluidic chip engagement tray incorporating a data transmitter/receiver, a microcontroller, and a user interface. In an embodiment, the data transmitter/receiver includes an RFID reader. In another embodiment, the transmitter/receiver detects the correct positioning of a microfluidic chip on the engagement tray.
In another embodiment, the instrument operates in association with a microfluidic chip comprising a data component.
In another embodiment of the invention, the instrument and microfluidic chip communicate with each other when the microfluidic chip is engaged in the instrument and the instrument is turned on.
According an embodiment of the invention, there is provided a programmable microfluidic chip comprising an inlet, microchannels, an outlet, and a data component.
In another embodiment of the invention the microfluidic chip of claim 1, wherein the data component is a radiofrequency identification tag (“RFID”). In other embodiments, the RFID has a defined readable range. The range is, in some embodiments, from 0 to 50 mm. In other embodiments, the range is 0 to 20 mm. In other embodiments, it is 0 to 5 mm. In another embodiment of the invention, the microfluidic chip is provided with a removable fitted manifold and cover.
In another embodiment of the invention, the data component includes stored data which is readable by, and which directs the behaviour of, an instrument for mixing. In another embodiment of the invention, the stored data includes a status indicator includes historical data of said microfluidic chip.
In another embodiment of the invention, the stored data includes the type or purpose of microfluidic chip.
In another embodiment of the invention, the data component is read by an instrument for mixing, processed by a microcontroller within said instrument, and a corresponding message is communicated to a user via a user interface on the instrument.
In another embodiment of the invention, the data read from the data component dictates what information the instrument sends to the user interface.
In another embodiment of the invention, the data read from the data component of the microfluidic chip contains information which is sent to the user interface and appears to a user as a set of instructions. In another embodiment, the information appears to the user as a set of options.
In another embodiment of the invention, the data component is capable of receiving, storing and emitting data.
According an embodiment of the invention, there is provided a system for formulating a therapeutic agent for research use, the system including an instrument having a pump, a microfluidic chip engagement tray incorporating a data transmitter/receiver, a microcontroller, a memory storage device, and a user interface, as well as an interchangeable microfluidic chip, and wherein the therapeutic agent is selected from the group consisting of nucleic acids, peptides, protein, or hydrophobic small molecules.
Other aspects and features of the present invention will become apparent to those ordinarily skilled in the art upon review of the following description of specific embodiments of the invention in conjunction with the accompanying figures.
In drawings which illustrate embodiments of the invention,
In accordance with a first embodiment of the invention, there is provided an microfluidic mixing instrument as generally shown in
As illustrated best in
Connected to main PCB 340 via ribbon cable connector 68 is reader 110, and the secondary PCB 112. Secondary PCB 112 operates the LEDs associated with m-chip entry 82. Also connected to secondary PCB 112 is cartridge switch 67 and start switch 66. Cartridge switch 67 is engaged to reader 110. Cartridge switch 66 is engaged to start button 87.
Secondary PCB 112 is normally within the base of instrument 100, below platform 115. Turning now to
An on/off power switch is located at the back of the instrument 100 in preferred embodiments.
The data emitting sensor 20 interacts with the instrument 100 reader 110 to accomplish the processes, checking for cartridge, whether used or not, informing user if not, whether the cartridge is compatible with the instrument 100, loading a recipe, prompting user for confirmation, executing process if user indicates yes, indicating success or error outcome, and recording usage data to the tag.
The microcontroller 300 (for example, a microcontroller such as an Atmel™ ATmega2560™ microcontroller, available from any robotics vendor including http://www.canadarobotix.com, www.BC-Robotics.com, https://www.buyapi.ca and many others) coordinates and controls the commands and feedback from and to the other components. In embodiments, microcontroller 300 is a single-board microcontroller used for building digital devices and interactive objects that can sense and control real objects. Main PCB 340 receives and delivers the commands and feedback to and from the other components. Microcontroller 300 is generally placed in front of the PCB 340 and behind the instrument front face 95.
There are one or two motors in preferred embodiments. These are shown only in block diagram form in
Data Component
In accordance with a second embodiment of the invention, there is provided an m-chip 50 according to the invention as exemplified by embodiments shown in
M-chip 50 is a solid material, such as rigid or semi-rigid plastic, metal or glass, and is manufactured to have inlet 55, microchannels with micromixing geometries, outlet 45, and a data component 20. M-chip 50 possesses strength and surfaces for a clamp to secure it into position and to allow an inlet pump 47 (not shown) to seal to inlet 55, in some embodiments in the form of a custom fitted lid that is placed on the m-chip 50 after reagents are added to inlet 55. In some embodiments, m-chips have a side flange 52 for stability and user manipulation. These are not necessary for the operation of the m-chip, but are added for convenience for the user. When correctly positioned in the instrument 100, m-chip 50 microchannels are hydraulically connected to the instrument pump(s) 47 which impels the flow of reagents from inlet 55 into microchannels via positive displacement, or by controlled pressurization of the inlet reservoirs 55 integrated within the chip 50 (shown for example in
As referred to above, the M-chip 50 in
Hydraulics
Syringe pumps are used in one embodiment, one per inlet. In some embodiments, there are two inlets 55 and there are engaged by separate pumps so that reagents from each inlet 55 are separately driven (at different speeds, for example).
A microchannel is defined as a channel with a hydraulic diameter of below 1mm. “Mixing geometries” are known in the art, and include herringbone and other patterns of microchannels, examples of which are disclosed in United States Patent Publication Nos. US20120276209A1, US20160214103A1, US20160235688A1, and PCT publication No. WO2016138175A1. In some embodiments, the data component 20 rests in a tag recess 25 in the m-chip 50 to reduce the risk of tampering or breakage. “Mixing” is meant to include any action wherein two or more materials are combined.
Data Emitting Sensor
Data emitting sensor 20 is embedded or adhered to an m-chip 50, has a sensitive range 80, and interacts with a data receiver 110. Sources for data emitting sensors are any electronics vendors online. The tags 20 are programmed using simple computer language and installed in m-chip 50 by hand, in some embodiments. In alternate embodiments, the tag 20 is manufactured into the m-chip and programmed after manufacture. In another embodiment the tag 20 is pre-programmed and manufactured into the m-chip after that.
In preferred embodiments, data emitting sensor 20 is an RFID tag.
Generally, RFID tags or radiofrequency identification tags are embedded with a transmitter and a receiver. The RFID component according to embodiments of the tags 20 has a chip that stores and processes information, and an antenna to receive and transmit a signal. The tag encodes the unique serial number for a specific m-chip 50, and certain characteristics can be programed in.
The RFID tags 20 used in some embodiments of the invention are passive, in that they use the reader's radio wave energy to relay their stored data back to the reader 110. In other embodiments, a powered RFID tag is embedded with a small battery that powers the relay of information.
The interaction of tag 20 with microfluidic mixing instrument as shown generally in
The interaction between tag 20 and reader 110 is unidirectional is some embodiments, or bidirectional in preferred embodiments.
In one embodiment, data emitting sensor 20 may include data in the form of an integer count, binary flag, defined character, string or equivalent to indicate whether or not it has been previously used and if so, how many times it has been used or how many uses remain. Such an application is valuable at occasions when the number of uses, including single-use, of an m-chip 50 requires enforcement for either regulatory or licensing reasons. Specifically in microfluidics, the presence of small channels which cannot be easily visualized may result in a condition where an m-chip 50 appears “clean” and usable to an operator, but in fact small amount of material (such as nucleic acids, salts, proteins or other molecules) representing cross-contamination between runs may be present. Additionally, use of the m-chip 50 may cause microscopic damage, not apparent to the operator, which would compromise future experiments carried out on said device 50.
In one embodiment, the m-chip 50 may contain a means of storing a set of instructions for the instrument to carry out on an inserted m-chip 50. This may include instrument settings such as temperatures, delay times, pressure values or any other parameter which could conceivably be incorporated onto an electromechanical instrument. In one embodiment, after an m-chip 50 is inserted into the instrument the recipe would be read and either executed (with or without a user prompt or warning). Such a configuration would be attractive in many scenarios. In one situation, a manufacturer may provide m-chip 50s as part of a larger kit where different kits may perform different tasks using the same m-chip. In this situation, a stored recipe allows the manufacturer to produce only one type of m-chip 50 but load a different recipe depending on which kit it will be bundled with. This approach reduces the likelihood of the error versus one where the operator would have to enter or select a recipe on the instrument. Additionally, this approach allows the manufacturer to update recipes or release new ones without having to perform updates to instruments deployed in the field. In a further embodiment, the m-chip 50 may contain multiple recipes, each with a corresponding signature which is recognized by the instrument, thus enabling the m-chip 50s to be backwards and cross-compatible with instruments containing different software or hardware versions.
Thus, in embodiments, recording usage data can be achieved with a writeable RFID tag 20 containing a defined section of memory with a flag which the instrument 100 may switch on to indicate that the m-chip 50 has been used. If re-use is permissible for a certain number of times under a particular licensing or regulatory condition, a block of memory is used to store how many more times the m-chip 50 may be used or how many uses remain.
In embodiments of the invention, the RFID tag 20 contains a memory for storing a set of instructions for the instrument to carry out on an inserted m-chip 50. This may include instrument settings such as temperatures, delay times, pressure values, flow rates, speed or distance of movement of actuators, or any other parameter which could conceivably be incorporated onto an electromechanical instrument. In one embodiment, after an m-chip 50 is inserted into the microfluidic mixing instrument, the recipe would be read and executed with or without further user action. In such an embodiment, a cartridge may be provided with one or more embedded or pre-loaded reagents for which the corresponding recipe may be programmed onto the cartridge to avoid error and to simplify the workflow of the operator.
This approach reduces the likelihood of user error. Additionally, the system of the invention allows the manufacturer to update recipes or release new ones without having to perform updates to instruments deployed in the field. In a further embodiment, the m-chip 50 may contain multiple recipes, each with a corresponding signature which is recognized by the instrument 100, even enabling the m-chips 50 to be backwards and cross-compatible with instruments containing different software or hardware versions
In certain embodiments, the instrument 100 may record data onto the m-chip 50. In one embodiment, if a fault occurs during operation, the instrument records items such as error codes, instrument settings, sensor readings etc. onto the m-chip. In this way, if the m-chip 50 is presented to the manufacturer or their representative, the information is read in order to diagnose the fault.
In one embodiment, a specially programmed m-chip 50 contains data to update the settings, parameters or other information on the instrument. In such an embodiment, once the m-chip 50 is read, the data on the instrument will be updated to the new value for future uses with standard microfluidic cartridges.
In one embodiment, the instrument may adapt its behaviour based on the information read from the m-chip 50. Different recipes or settings present on the m-chip 50 may require different interfaces, options, parameters, indicators etc. to be presented to the operator. In a further embodiment, the chip may contain data which is or is used to generate steps for the operator to follow (for example, what volumes to load onto the chip) such that is the operator taps the chip on the instrument, the instrument guides the operator through the steps of a recipe.
In some embodiments, the reader 110 is a two-way radio transmitter-receiver, whose location in the Instrument is indicated in
For example, when instrument is switched on (power switch on rear of machine), and when an m-chip 50 is inserted into chip entry 82 along platform 115, a pressure sensor on platform 115 provides a signal to cartridge switch 67, which signals the reader 110 to emit a signal to the tag 20 using a built in antenna.
Correct placement and orientation of m-chip 50 is guided by the pressure sensor primarily, then by the tag 20 interacting with reader 110 for fine tuning, which is dependent on the specific range of the data signal range 80 as illustrated in
The tag 20 responds to reader 110 with the information written in tag 20 memory.
The logical pattern embodied in the m-chip and instrument of the invention, and their interaction with each other, is illustrated in
If the m-chip is not detected on platform 115:
or if m-chip is detected on platform 115, but not properly positioned:
The menu screen allows the user to select a mode.
If an m-chip is not the right type for the MODE selected by user on the GUI 90,
If the m-chip has been used already:
The exact wording can be updated in each chip manufacture. It should be noted that this is a great advance over the prior art instrument and microfluidic chip use, during which information was not available to user on the success or failure of a chip and its formulation.
The instrument 100 selects the appropriate information to display on its GUI 90 based on data emitting sensor 20 data.
Thus this disclosure is directed towards a disposable cartridge containing an m-chip 50 and an embedded data emitting component, such as an RFID tag 20, to store data. By using this cartridge with an accompanying scientific instrument, together referred to as a system, information can be transferred in both directions (read from the m-chip 50 to the instrument or written by the instrument onto the m-chip 50 at tag 20) to facilitate ease of use, software updates, troubleshooting, and end user licensing among other tasks. Various embodiments are described below which is used independently or combined to form further embodiments.
RFID 20 is used as means to store and read data on an m-chip 50. In one such instance, an RFID reader 110 was embedded inside of a NanoAssemblr™ Spark™ laboratory research instrument 100. In this specific case, a DLP-RFID2 (DLP Design, Allen, Tex.) reader (#+1) was attached to the underside of the instrument's microfluidic cartridge receiving tray 115. This reader was positioned such that a 7.5 mm RFID tag (meeting the specifications of ISO/IEC 15693. (Verigenics, Southampton, Pa.) installed on the front, underside of a Spark™ m-chip 50 was successfully read when properly inserted and positioned within the Spark. The RFID reader 110 was directly connected to the instrument's internal microcontroller (#+2) in such a way as to communicate using industry standard protocols.
During m-chip 50 manufacturing, the RFID tag 20 was affixed to a recess 25 on the front, underside of an m-chip 50 using a double-sided adhesive film. The RFID 20 was programmed using standard techniques for programming such a tag (method may vary, but vendors provide standard instructions or software). A simple handheld programming device is used to program the tags in this example.
The m-chip 50 was programmed to run a specific set of parameters required to formulate 2 nmol of siRNA into lipid nanoparticles for delivery to neurons in vitro.
Data to be stored on the RFID tag was loaded onto a host computer in the form of a .csv file. The host computer broke this data into 8-byte blocks that were written to an RFID tag one at a time. The host computer sent a Write Block command, with one block of data, to the RFID read/write module via an RS-232 serial connection. The RFID read/write module generated an electromagnetic field to power and communicate with an RFID tag according to the ISO-15693 standard.
The module sent out the Write Block command. If an RFID tag was within range of the module's antenna, the tag stores the block of data into non-volatile internal memory and responds to the RFID module with a Success code. The RFID module waited for a tag response, and then reported back to the host computer about whether the write was successful.
Steps 4-7 were repeated until all of the data was sent to the RFID tag and stored.
A pass-fail scenario that was programmed into the chip provided error codes simply stating the following:
ST01: no chip inserted.
ST02: used chip inserted
ST03: wrong chip for current mode, kit chip in formulation mode
ST04: wrong chip for current mode, formulation chip in kit mode
ST05: chip inserted while purging
ST06: can't read chip, no RFID header
ST07: can't read chip, bad checksum
To formulate using a Spark™ mixing instrument, the operator used an m-chip as shown in
The instrument then ran the formulation as per the parameters stored on the RFID tag. After the formulating process was successfully completed, the data on the tag was updated by the reader in theSpark™ to indicate that the m-chip had been used.
After formulation, the operator removed the m-chip from the instrument, removed the cap and manifold and pipetted out the resultant formulation from the outlet well. The m-chip 50 (with corresponding tag) was then disposed of according to local regulations.
An m-chip is prepared as in Examples 1 and 2, but error codes include:
ST08: flow rate error
ST09: pressure error
If an error had occurred during formulation, such as a loss of pressure, a corresponding error code or message would have been displayed to the operator and written to the RFID tag.
Enhanced feedback from Instrument to tag, and vice versa, includes pressure losses, unexpected resistance, or unexpected lack of resistance, These enhanced data are included in the GUI readout to inform user of further exceptions. These exceptions can help diagnose mechanical issues with the instrument, which will help with repair of the instrument.
While specific embodiments of the invention have been described and illustrated, such embodiments should be considered illustrative of the invention only and not as limiting the invention as construed in accordance with the accompanying claim
| Number | Date | Country | |
|---|---|---|---|
| 62359123 | Jul 2016 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | 16314941 | Jan 2019 | US |
| Child | 17340863 | US |
