Patent Application
20170368365
This invention relates to assaying the application of energy to samples. More specifically, this invention relates to a device for applying various amounts of energy, such as electromagnetic energy, to different wells of a culture plate.
Applying electromagnetic energy to living cells and tissues has been shown to have biological effects on the cells and tissues. Various types of energy have been used for treatment, such as non-thermal plasma, radio frequency, light, electrical fields, and magnetic fields. Pulsing, or modulating, the energy at different frequencies can change the effect of the treatment. However, clinical studies for determining optimum frequencies for specific treatments have been difficult to justify because the physical mechanism behind the biological interaction is not well understood. Clinical studies are also very expensive and the protocol for selecting an optimum frequency is poorly defined.
Conventionally, in vitro studies have been used to determine an optimal dosage of a medicament that causes a desired outcome. In one approach, cells or tissue samples are deposited in a multi-well culture plate and each well is dosed with a different amount of the medicament. The dosage is preferably applied with a programmed robotic device to increase speed and accuracy of the testing. The cells or tissue samples of each well may then be individually analyzed to determine the effects of the applied dosage.
While many methods and devices exist for studying medicament dosages applied to cells or tissue samples, little exist for studying energy applied to cells or tissue samples in an inexpensive and accurate manner. Thus, it would be desirable to have a device for performing in vitro studies with various types of energy.
Some embodiments of the invention provide an assay device for a culture plate including a plurality of culture wells. The assay device includes a housing, an array, and a control module. The housing includes a socket sized to receive the culture plate. The array includes a plurality of emitters and is positioned within the housing adjacent to the socket so that the plurality of emitters are each aligned with at least one of the culture wells when the culture plate is received in the socket. The control module is operably coupled to the array and is configured to individually drive the plurality of emitters. The control module is configured to drive at least two of the plurality of emitters at different frequencies simultaneously.
Some embodiments of the invention provide a method of assaying the application of energy to a plurality of samples in vitro. The method includes applying the plurality of samples to individual wells of a culture plate and aligning at least one energy emitter with each individual well of the culture plate. The method also includes applying energy at a separate frequency toward each individual well via each of the at least one energy emitters for a first time period and incubating the wells for a second time period.
Before any embodiments of the disclosure are explained in detail, it is to be understood that the invention is not limited in its application to the details of construction and the arrangement of components set forth in the following description or illustrated in the following drawings. The invention is capable of other embodiments and of being practiced or of being carried out in various ways. Also, it is to be understood that the phraseology and terminology used herein is for the purpose of description and should not be regarded as limiting. The use of “including,” “comprising,” or “having” and variations thereof herein is meant to encompass the items listed thereafter and equivalents thereof as well as additional items. Unless specified or limited otherwise, the terms “mounted,” “connected,” “supported,” and “coupled” and variations thereof are used broadly and encompass both direct and indirect mountings, connections, supports, and couplings. Further, “connected” and “coupled” are not restricted to physical or mechanical connections or couplings.
The following discussion is presented to enable a person skilled in the art to make and use embodiments of the disclosure. Various modifications to the illustrated embodiments will be readily apparent to those skilled in the art, and the generic principles herein can be applied to other embodiments and applications without departing from embodiments of the invention. Thus, embodiments of the invention are not intended to be limited to embodiments shown, but are to be accorded the widest scope consistent with the principles and features disclosed herein. The following detailed description is to be read with reference to the figures, in which like elements in different figures have like reference numerals. The figures, which are not necessarily to scale, depict selected embodiments and are not intended to limit the scope of embodiments of the invention. Skilled artisans will recognize the examples provided herein have many useful alternatives and fall within the scope of embodiments of the invention.
Generally, embodiments of the present disclosure provide an assay device that enables treating multiple wells of a culture plate simultaneously with different pulse frequencies of energy. The device may receive the culture plate in a socket so that the plurality of wells are aligned in close proximity to an array of energy emitters, each of which emits a different pulse frequency simultaneously. This enables many frequencies to be tested simultaneously, speeding up the search for a frequency that provides a desired outcome on samples in the culture wells.
As shown in
Accordingly, the array 24 can be positioned within the housing 12 adjacent to (e.g., below) the socket 14. As described in more detail below with respect to
According to some embodiments, the control module 28 can be operably coupled to the array 24 (e.g., through the array driver 26) and can be configured to individually drive each emitter 48 at a specific frequency for a set time period. For example, the control module 28 can control the array driver 26 to drive each emitter 48 at a different frequency, or to drive groupings of more than one emitter 48 at different frequencies. The control module 28 may include an application-specific integrated circuit (ASIC), a programmable logic controller (PLC), or, preferably, a field-programmable gate array (FPGA), such as the Xilinx Artix 7. A signal generator for each emitter 48 (e.g., a square wave signal generator) can be implemented in VHDL code as a phase accumulator, or numerically controlled oscillator (NCO). In a preferred embodiment, control module output to the array driver 26 consists of the most significant bit of the phase accumulator. Using only a single bit of the output may add deterministic phase jitter, but it allows the device to define individual emitter units (e.g., up to ninety-six units) with reasonable FPGA resources. Furthermore, using a 200 MHz clock and both edges of the clock, the peak timing uncertainty can be about ±2.5 nanoseconds (ns).
The control module 28 can also be operably coupled (e.g., electrically connected) to the memory card slot 38, the user interface 16, the RS-485 interface 40, the fan 34, the power supply 36 and/or other connections (not shown). More specifically, the memory card slot 38 can be sized to receive a memory device (such as a data card) allowing, for example, the control module 28 to upload control instructions and test data from the memory card and/or download test data to the memory card. Alternatively, in some embodiments, the memory card slot 38 can be sized to receive a USB flash drive. Also, in some embodiments, the control module 28 can include internal memory accessible via one of the I/O interfaces.
In some embodiments, the control module 28 can connect to an external computer via the RS-485 interface 40. The RS-485 interface 40 may also be used to connect multiple devices 10 (e.g., up to 128 devices) in a daisy-chain configuration, for example, using a MODBUS communication protocol. The interface can be designed so that the first device 10 in the cable chain the lowest address, and the subsequent devices 10 have sequentially higher addresses.
In some embodiments, the device 10 can include more than one RS-485 interface 40. Additionally, it should be noted that, while the memory card slot 38 and the RS-485 interface 40 are shown and described herein, it within the scope of this disclosure to include additional input/output interfaces and/or other communications circuits. For example, the device 10 can include a power supply interface (not shown), permitting the device 10 to be connected to and powered by an external power supply.
Accordingly, the control module 28 can receive instructions via the memory card slot 38 (i.e., via an inserted memory device), the RS-485 interface 40 (i.e., via a connected external computer), or additional input/output interfaces. Furthermore, the control module 28 can receive user inputs via the user interface 16. As shown in
Furthermore, the user interface 16 can include the display 20. The display 20 may be, for example, a light emitting diode (LED) display, an organic light emitting diode (OLED) display, a digital display, or another suitable display. In one embodiment, the display 20 is a 2×20-character OLED display. The control module 28 may control the display 20 to display to a user, for example, program status, on/off status, device status, present temperatures, a timer, test data, programs, or the like. In one example, the device 20 can display test data or programs in a list format, and a user can toggle the lists and/or select a desired program using the push buttons 18. Alternatively, the control module 28 may automatically execute a program upon receiving a memory card within the memory card slot 38 and being powered on.
Internally, the control module 28 can control operation of the fan 34 as part of a thermal control system of the device 10. More specifically, the thermal control system can include the fan 34, the heat sink 32, the thermal control devices 30 (such as thermoelectric coolers, TECs), and one or more temperature sensors (not shown). For example, the array 24, the array driver 26, and/or the control module 28 can be thermally coupled to the heat sink 32, the TECs 30, and the fan 34. The heat sink 32 may act as a passive cooling device, while the fan 34 may be operated for active cooling, and the TECs 30 may be operated for heating or cooling.
The control module 28 can control operation of the fan 34 and/or the TECs 30 based on a specific treatment program (e.g., the control module 28 may operate the device 10 at a desired temperature or within a desired temperature range based on received temperature control data for a specific program). For example, during and/or after energy application to the culture wells 42, the control module 28 can control the thermal control system (in particular, the fan 34 and the TECs 30) to incubate the culture wells 42 for a preset time period.
More specifically, the temperature sensors can be positioned on the array 24, the housing 12, and/or the heat sink 32 and can be in communication with the control module 28. The control module 28 can monitor the temperatures of these components (i.e., by retrieving a current temperature from one or more of the temperature sensors) and determine whether passive or active cooling is necessary or whether heating is necessary. That is, the control module 28 can operate the fan 34 based on input from one or more of the temperature sensors. In one embodiment, the fan 34 can draw in air from one or more bottom cutouts of the housing 12, past the heat sink 32 and up through the socket 14 (i.e., past the culture plate 44 received in the socket 14). Additionally, in some embodiments, the control module 28 may display an alarm (e.g., via the display 20) if a temperature of one of the components exceeds a high or low temperature threshold for a set time period despite heating or cooling.
Furthermore, it should be noted that other thermal control devices are contemplated within the scope of this disclosure. Additionally, the heat sink 32 and all internal electronics of the device 10 can be thermally insulated from the housing 14.
The following paragraphs describe specific arrays 24 for devices 10 configured to treat culture wells 42 with RF energy, electric fields, magnetic fields, light, and non-thermal plasma, respectively. For example, the above-described components may be common to each device 10, with the exception of the arrays 24 and array drivers 26, which may be specific to the type of energy emission of the device 10. Thus, some embodiments provide different build options for an RF-specific device, an electric field-specific device, a magnetic field-specific device, a light-specific device, and a non-thermal plasma-specific device. Alternatively, in some embodiments, a device 10 can include all the above common components and can be configured to receive different arrays 24 and array drivers 26 in an interchangeable manner. Thus, such a device 10 can be used for all types of energy emissions.
The RF driver board 26 can be a separate printed circuit board that drives the RF array 24, as shown in
The magnetic field array driver 26 can be a separate printed circuit board that drives the magnetic field array 24. In some embodiments, the duty cycle of the driving waveform can be adjusted to produce a DC bias of 2 milliamperes (mA) at 5 V. A circuit (not shown) for monitoring DC bias can be connected to a single element 48, on the assumption that all elements 48 will have a similar bias. A calibration routine can be used to determine the required duty cycle for a given frequency. Additionally, as shown in
The optical driver board 26 can be a separate printed circuit board that drives the optical array 24. The LED drivers 26 can be designed to run at a fixed current, but the average power can be reduced by adjusting the duty cycle. In some embodiments, the LEDs 48 can be mounted upside down into the driver PCB 26 to create a flush surface for thermal contact with the culture plate 44. Additionally, as shown in
The plasma array driver 26 can be a separate printed circuit board that drives the plasma array 24. In one embodiment, the plasma array driver 26 can require multiple lines from the control module 28 to control the plasma driver 26 and high voltage transformers 60 to create a total of twenty-four drivers 26. Each driver 26 can then be connected to a set of 4 micro-plasma arrays 24, which sit underneath the culture wells 42. Each set of plasma arrays 24 can present a capacitance of about 20 picofarads (pF). This capacitance resonates with the plasma drive transformer 60, and the plasma drive frequency can be about 420 kHz. In one embodiment, the plasma modulation frequency can be adjusted by the control module 28 up to 10 kHz. The plasma tends to run at a fixed current, and a duty cycle of 10% may be used to reduce average power and extend the array life. Additionally, as shown in
In light of the above, embodiments of the present disclosure provide a device 10 that enables treating multiple wells 42 of a culture plate 44 simultaneously with different pulse frequencies of energy, such as RF, electric field, magnetic field, light, or plasma.
More specifically, at step 70, a sample culture of interest, such as cells, tissues, bacteria, fungus or virus, is deposited in each well 42 of the culture plate 44. For example, each well may include the same type of sample, or different wells may include different types of samples. Next, at step 72, the culture plate 44 is inserted into the socket 14 of the device 10. As described above, the socket 14 can be sized so that the culture plate 44 rests within the socket 14 atop the array 24. Additionally, in some embodiments, steps 70 and 72 may be reversed (i.e., the culture plate 44 can be inserted into the socket 14 and then the samples can be deposited in each well 42).
When the culture plate 44 is received within the socket 14, a treatment program may be run on the device 10 at step 74. Generally, a treatment program can include specific frequencies to apply, the first set time period, the second set time period, temperature set points or thresholds, and/or other testing parameters. For example, the device 10 may be powered on and a memory card may be inserted in the memory card slot 38 so that the control module 28 automatically executes a treatment program stored on the memory card. In another example, the device 10 may display multiple treatment program options on the display 20 and a user may select a desired treatment program for the control module 28 to run via the push buttons 18. In yet another example, the device 10 may be connected to an external computer via the RS-485 interface 40 (or another input/output interface 22) and a user may select a desired treatment program for the control module 28 to run via the external computer.
According to the selected treatment program, the control module 28 can control the array 26 to apply energy to the culture plate 44 at step 76. More specifically, the control module 28 can generate a set of separate frequencies (i.e., more than one frequency) for each emitter 48 or for groups of emitters 48 to emit toward individual wells 42 of the culture plate 44 simultaneously. As discussed above, the array 24 can be configured so that a single emitter 48 is aligned under each well 42, or so that groups of emitters 48 are aligned under each well 42. Thus, each emitter 48 (or each group of emitters 48) can output a different frequency simultaneously in order to test the effects of multiple frequencies on separate samples (i.e., in the separate culture wells 42) at the same time. In one example, the control module 28 can drive at least two emitters 48 at different frequencies simultaneously. In another example, the control module 28 can generate a set of ninety-six separate frequencies simultaneously to individual control ninety-six emitters 48. The control module 28 can apply the energy to the culture wells 42 for a first set time period according to the selected treatment program.
The control module 28 can also operate the thermal control system to incubate the culture wells 42 for a second set time period, at step 78, according to the selected treatment program. For example, in one embodiment, the control module 28 can thermally control the temperature of the array 24 in order to incubate the wells. In some embodiments, the second set time period may be equal to the first set time period of step 76. Additionally, in some embodiments, the second set time period may partially or completely overlap with the first set time period.
Once the second set time period is completed, the culture wells 42 may be analyzed to determine the effects of the applied energies to the samples, at step 78. In some embodiments, the device 10 may automatically turn off once the second set time period is completed, may automatically save treatment data to memory (e.g., store frequency and temperature control data to internal memory or the memory device), or may display an alert via the display 20 to notify a user that the treatment program is complete.
In accordance with the above method, multiple samples can be simultaneously treated in vitro with different frequencies and examined to determine which sample had a desired or optimal outcome, thus allowing a determination of the optimal frequency for a specific treatment. Furthermore, the method may be repeated, for example, to see if the energy effects are repeatable. For example, the control module 28 can apply the same treatment program, but shuffle the modulation frequencies to different emitters 48. In particular, repeating the methods and shuffling the frequencies can compensate for signal overlap between adjacent array elements. Additionally, the present methods can be applied to multiple devices 10 simultaneously using a daisy-chain configuration, as described above. Multiple tests can thus be completed quickly and accurately and both frequency and temperature data may be stored, for example, to memory devices or a database as part of a research program and to track tested frequencies.
In light of the above, embodiments of the disclosure provide a device and method for studying the application of energy at different frequencies to samples in vitro. By allowing multiple samples to be tested at different frequencies simultaneously and then incubated, the present device and method provide a standalone low-cost, quick, and accurate way to study the biological effects of samples based on the modulation frequency of RF, electric fields, magnetic fields, light, and plasma.
It will be appreciated by those skilled in the art that while the invention has been described above in connection with particular embodiments and examples, the invention is not necessarily so limited, and that numerous other embodiments, examples, uses, modifications and departures from the embodiments, examples and uses are intended to be encompassed by the claims attached hereto. The entire disclosure of each patent and publication cited herein is incorporated by reference, as if each such patent or publication were individually incorporated by reference herein. Various features and advantages of the invention are set forth in the following claims.
This application claims priority under 35 U.S.C. §119 to U.S. Provisional Patent Application No. 62/355,543 filed on Jun. 28, 2016, the entire contents of which is incorporated herein by reference.
| Number | Date | Country | |
|---|---|---|---|
| 62355543 | Jun 2016 | US |
