Substrate for performing quantization process and related systems and methods

Abstract

Substrates and related systems and methods for performing quantization processes are disclosed. In particular implementations, an apparatus includes a substrate and an imaging system. The substrate includes a pair of plates and a plurality of spacers positioned between the plates to define a gap between the pair of plates. A portion of a sample is to be received within the gap of the substrate, and the imaging system is to obtain image data of the portion of the sample. The image data is to be used to determine a concentration of the portion of the sample.

Description

[0001] Related patent applications

[0002] This application claims the benefit of and priority to U.S. Provisional Patent Application No. 63 / 355,879, filed on June 27, 2022, the contents of which are incorporated herein by reference in their entirety and for all purposes. Background Art

[0003] DNA libraries can be prepared to allow sequencing of the samples. Summary of the invention

[0004] By providing a library preparation system and method, the shortcomings of the prior art can be overcome, and the benefits described later in this disclosure can be achieved. Various specific implementations of the apparatus and method are described below, and these apparatus and methods (including and excluding the additional specific implementations listed below) can overcome these shortcomings and achieve the beneficial effects described herein in any combination (provided that these combinations are not inconsistent).

[0005] In a first embodiment, an apparatus includes a substrate and an imaging system. The substrate includes a pair of plates and a plurality of spacers positioned between the plates to define a gap between the pair of plates. A portion of a sample is to be received within the gap of the substrate, and the imaging system is to obtain image data of the portion of the sample. The image data is to be used to determine a concentration of the portion of the sample.

[0006] In a second specific implementation, a method includes: dispensing a sample onto a first plate of a substrate; positioning a second plate of the substrate on the first plate and using a plurality of spacers to define a gap between the first plate and the second plate; obtaining image data of the sample; and using the image data to determine a concentration of the sample.

[0007] In a third specific implementation, a method includes: dispensing a sample in an inlet of a channel of a substrate; obtaining image data of the sample; using the image data to determine a concentration of the sample, and causing the sample to flow out of an outlet of the channel.

[0008] In a fourth embodiment, a device includes a substrate and an imaging system. The substrate includes a channel, the channel includes an inlet and an outlet, and a valve. The outlet is in fluid communication with the channel. The valve is disposed between the inlet and the outlet. The channel includes a first portion between the valve and the inlet and a second portion between the valve and the outlet. A portion of a sample is to be received in the first portion of the channel of the substrate, and the imaging system is to obtain image data of the portion of the sample. The image data is to be used to determine a concentration of the portion of the sample.

[0009] In a fifth embodiment, an apparatus includes a substrate, the substrate including a channel and a valve, the channel including an inlet and an outlet, the valve being disposed between the inlet and the outlet. The channel includes a first portion between the valve and the inlet and a second portion between the valve and the outlet. A portion of a sample is to be received in the first portion of the channel of the substrate, and an imaging system is to obtain image data of the portion of the sample. The image data is to be used to determine a concentration of the portion of the sample.

[0010] In a sixth embodiment, a device includes a circular sub-state, the circular sub-state including a plurality of channels. Each channel includes an inlet, an outlet, and a valve disposed between the inlet and the outlet. The channel includes a first portion located between the valve and the inlet and a second portion located between the valve and the outlet.

[0011] In a seventh specific implementation, a method includes: rotating a substrate around a rotation axis at a first rate to draw a portion of a sample into a first portion of a channel of the substrate. The substrate includes the channel, an inlet in fluid communication with the channel, an outlet, and a valve, the valve being disposed between the inlet and the outlet. The channel includes the first portion between the valve and the inlet and a second portion between the valve and the outlet. The method includes: stopping the rotation of the substrate; and obtaining image data of the portion of the sample. The image data is to be used to determine the concentration of the portion of the sample. The method includes: rotating the substrate around the rotation axis at a second rate to push the portion of the sample through the valve and into the second portion of the channel of the substrate and out of the outlet.

[0012] In an eighth specific implementation, a method includes: rotating a substrate around a rotation axis to draw a portion of a sample into a first portion of a channel of the substrate. The substrate includes the channel, an inlet, an outlet in fluid communication with the channel, and a valve disposed between the inlet and the outlet. The channel includes the first portion between the valve and the inlet and a second portion between the valve and the outlet. The method includes: stopping the rotation of the substrate; and obtaining image data of the portion of the sample. The image data is to be used to determine the concentration of the portion of the sample. The method includes: actuating the valve to an open position; and rotating the substrate around the rotation axis to push the portion of the sample through the valve and into the second portion of the channel of the substrate and out of the outlet.

[0013] Further according to the foregoing first specific implementation, second specific implementation, third specific implementation, fourth specific implementation, fifth specific implementation, sixth specific implementation, seventh specific implementation and / or eighth specific implementation, a device and / or a method may also include or comprise any one or more of the following:

[0014] In one implementation, the spacer includes a ball bearing having opposing flat surfaces.

[0015] In another specific implementation, the plurality of spacers includes three spacers.

[0016] In another implementation, the gap is about 500 microns.

[0017] In another specific implementation, the device includes a channel defined between the pair of plates.

[0018] In another specific implementation, the base plate further includes a seal positioned between the pair of plates and defines the channel.

[0019] In another implementation, the seal is positioned around a perimeter of the pair of plates.

[0020] In another implementation, the substrate includes a mask.

[0021] In another implementation, the mask is an opaque mask.

[0022] In another specific implementation, the substrate includes an inlet and an outlet in fluid communication with the gap.

[0023] In another implementation, the substrate includes a plurality of channels.

[0024] In another specific implementation, the substrate includes a first inlet, a second inlet, a third inlet, and an outlet in fluid communication with the gap.

[0025] In another implementation, the spacer includes a ball bearing having opposing flat surfaces.

[0026] In another specific implementation, the plurality of spacers includes three spacers.

[0027] In another specific implementation, the method further includes positioning a seal around a perimeter of the first plate and the second plate.

[0028] In another specific implementation, obtaining the image data includes obtaining the image data through a window of the substrate.

[0029] In another specific implementation, the window includes an unmasked area, and the masked area can surround the unmasked area.

[0030] In another specific implementation, the method includes dispensing a fluid into the channel to separate the sample from another sample.

[0031] In another specific implementation, the fluid includes oil.

[0032] In another specific implementation, dispensing the fluid into the channel includes dispensing the fluid into a second inlet of the channel.

[0033] In another implementation, the device includes a pump for moving the sample from the inlet to the outlet.

[0034] In another specific implementation, the pump includes a plate including a rotational axis and an actuator for rotating the plate, the base plate being coupled to the plate and extending radially from the rotational axis.

[0035] In another implementation, the apparatus includes a second substrate coupled to the plate and extending radially from the axis of rotation.

[0036] In another specific implementation, the plate includes a first position and a second position. The substrate is positioned at the first position, and the second substrate is positioned at the second position. The first position is opposite to the second position.

[0037] In another implementation, the second substrate is a mirror image of the substrate on the plate.

[0038] In another specific implementation, the pump includes a light access region.

[0039] In another implementation, the device includes a waste reservoir fluidly coupled to the outlet.

[0040] In another implementation, the waste reservoir includes a vent.

[0041] In another specific implementation, the vent includes porous frit.

[0042] In another specific implementation, the waste reservoir includes a disposable microfluidic card assembly.

[0043] In another specific implementation, the valve includes a virtual valve.

[0044] In another specific implementation, the virtual valve includes a constriction.

[0045] In another specific implementation, after obtaining the image data of the portion of the sample, the portion of the sample is flowed from the first portion through the valve, the second portion, and the outlet.

[0046] In another specific implementation, the valve includes a virtual valve.

[0047] In another specific implementation, the virtual valve includes a constriction.

[0048] In another specific implementation, the first portion of the channel has a first width, the second portion of the channel has a second width, and the virtual valve has a third value, the third width being smaller than the first width and the second width.

[0049] In another implementation, the channel has a height between about 200 microns and about 500 microns.

[0050] In another implementation, the substrate includes a mask.

[0051] In another implementation, the mask is an opaque mask.

[0052] In another implementation, the substrate includes a microfluidic card.

[0053] In another specific implementation, the valve includes an elastomeric valve or a rotating face seal valve.

[0054] In another specific implementation, the device includes an inlet port at the inlet, the inlet port being used to funnel the portion of the sample into the channel.

[0055] In another specific implementation, the inlet port includes a flow channel, and the flow channel includes a frusto-conical flow channel.

[0056] In another specific implementation, the inlet port includes a first surface and a second surface. The first surface is coupled to the substrate. The truncated cone-shaped flow channel includes a base opening at the first surface and a top opening at the second surface. The base opening has a periphery. The inlet is tangent to the base opening.

[0057] In another implementation, the circular substrate comprises a plastic disk.

[0058] In another specific implementation, the circular substrate includes a circular wafer.

[0059] In another specific implementation, the circular substrate includes a rotation axis, and the device includes an actuator for rotating the circular substrate around the rotation axis.

[0060] In another specific implementation, the method includes receiving the portion of the sample in a waste reservoir fluidly coupled to the outlet.

[0061] In another specific implementation, the method includes flushing the channel by flowing a flushing fluid through the channel and between the inlet and the outlet.

[0062] In another specific implementation, the valve is a virtual valve.

[0063] In another specific implementation, the virtual valve includes a constriction.

[0064] In another specific implementation, the method includes flushing the channel by flowing a flushing fluid through the channel and between the inlet and the outlet.

[0065] It should be understood that all combinations of the foregoing concepts and the additional concepts discussed in more detail below (provided that these concepts are not mutually inconsistent) are contemplated as being part of the subject matter disclosed herein and / or can be combined to achieve the specific benefits of the specific aspects described herein. In particular, all combinations of the claimed subject matter appearing at the end of this disclosure are contemplated as being part of the subject matter disclosed herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0066] Figure 1 A schematic diagram of a specific implementation of a system according to the teachings of the present disclosure is shown.

[0067] Figure 2 It can be used to implement Figure 1 A top isometric view of a specific implementation of a substrate substrate.

[0068] Figure 3 yes Figure 2 Cross-sectional view of the substrate.

[0069] Figure 4 yes Figure 2 A cross-sectional view of a substrate showing a first plate removed from a second plate and a contact dispenser dispensing the parts onto the second plate.

[0070] Figure 5 yes Figure 2 A cross-sectional view of a substrate showing a first plate positioned on top of a second plate and an imaging system acquiring image data of one of the portions.

[0071] Figure 6 It can be used to implement Figure 1 A top isometric view of a specific implementation of a substrate substrate.

[0072] Figure 7 yes Figure 6 A cross-sectional view of a substrate showing a first plate positioned on top of a second plate and an imaging system acquiring image data of the portion.

[0073] Figure 8 It can be used to implement Figure 1 A cross-sectional view of a specific implementation of a substrate.

[0074] Fig. 9 It can be used to implement Figure 1 A top isometric view of a specific implementation of a substrate substrate.

[0075] Fig.10 yes Fig. 9A cross-sectional view of a substrate showing a first plate positioned on top of a second plate and an imaging system acquiring image data of a portion aligned with a window of the substrate.

[0076] Fig.11 Illustrated description for use Figures 1 to 10 or a flowchart of a process for a substrate and imaging system of any implementation in any of the other implementations disclosed herein.

[0077] Fig.12 Illustrated description for use Figures 1 to 10 or another flow chart of a process for a substrate and imaging system of any implementation in the other implementations disclosed herein.

[0078] Fig.13 It can be used to implement Figure 1 Schematic implementation of a substrate and imaging system.

[0079] Fig.14 yes Fig.13 Top view of the base plate and pump.

[0080] Fig.15 yes Fig.13 Detailed view of the baseplate and part of the pump.

[0081] Fig.16 It can be used to implement Figure 1 Schematic implementation of a circular base plate and a pump.

[0082] Fig.17 A flow chart of a method for implementing the examples disclosed herein is illustrated.

[0083] Fig.18 Another flow chart for implementing a method of the examples disclosed herein is illustrated. DETAILED DESCRIPTION

[0084] Although the following text discloses a detailed description of a specific implementation of the method, apparatus and / or article of manufacture, it should be understood that the legal scope of the property rights is defined by the text of the claims set forth at the end of this patent. Therefore, the following detailed description should be understood to be merely exemplary and does not describe every possible specific implementation, because it is impractical, if not impossible, to describe every possible specific implementation. Many alternative specific implementations can be implemented using current technology or technology developed after the filing date of this patent. It is contemplated that such alternative specific implementations will still fall within the scope of the claims.

[0085] Figure 1A schematic diagram of an implementation of a system 100 according to the teachings of the present disclosure is shown. For example, the system 100 can be used to automatically, easily, and efficiently perform a quantification process used in preparing a DNA library for sequencing applications. In the illustrated implementation, the system 100 includes a workspace 106. In the illustrated implementation, the workspace 106 includes a contact dispenser 145, a stage 148, and an analyzer area 154. The stage 148 can be an xy stage.

[0086] The stage 148 has a plate receptacle 160 that receives a plate 161, and the analyzer region 154 includes a base plate 162 and an imaging system 164. The plate receptacle 160 may be referred to as a plate station. The imaging system 164 may be a fluorescence imaging system, a fluorescence spectrophotometer including an objective lens and / or a solid-state imaging device. The solid-state imaging device may include a charge coupled device (CCD) and / or a complementary metal oxide semiconductor (CMOS).

[0087] The system 100 also includes a drive assembly 173 and a controller 176. The plate 161 may contain different samples 165. The samples 165 may be biological samples derived from humans, animals, plants, bacteria, or fungi. Other sources of obtaining biological samples may prove suitable. The stage 190 may be coupled to the contact dispenser 145 and used to move the contact dispenser. The stage 190 may be a z stage. Alternatively, the stage 190 may be omitted.

[0088] The analyzer region 154 of the system 100 can be used to perform a quantification process. In the illustrated implementation, the substrate 162 of the analyzer region 154 is carried by the stage 148, and the imaging system 164 is spaced apart from the stage 148 and coupled to a portion of the system 100, such as a frame of the system 100. The imaging system 164 may alternatively be carried by the stage 148. The substrate 162 is shown to include a pair of plates 200, 202, with a gap 204 defined between the pair of plates. The plate 200 may be referred to as a first plate or alternatively as a second plate, and the plate 202 may be referred to as a second plate or alternatively as a first plate. The substrate 162 also has an inlet 206 and an outlet 208 in fluid communication with the gap 204, and a seal 210 positioned between the pair of plates 200, 202. The plates 202, 202 and the seal 210 define a channel 212 located between the inlet 206 and the outlet 208. The waste reservoir 214 is fluidly coupled to the outlet 208 of the base plate 162 via a fluid line 216. A pump 215 may be fluidly coupled between the outlet of the base plate 162 and the waste reservoir 214. The pump 215 may be implemented by a syringe pump, a peristaltic pump, and / or a diaphragm pump. However, other types of fluid transfer devices may be used.

[0089] The stage 148 aligns the contact dispenser 145 with the plate 161 to perform the quantification process, and the contact dispenser 145 aspirates a portion of the reagent 178 and the sample 165 from the well 180 of the plate 161. The portion of the reagent 178 and the sample 165 may be about 2 μL.

[0090] The stage 148 aligns the contact dispenser 145 with the inlet 206 of the substrate 162, and the contact dispenser 145 dispenses the reagent 178 and the portion of the sample 165 into the inlet 206 of the substrate 162. In this implementation, the reagent 178 and the portion of the sample 165 can flow between the inlet 206 and the outlet 208 and / or be positioned therebetween, and the imaging system 164 obtains image data of the reagent 178 and the portion of the sample 165. The imaging system 164 and / or the system 100 uses the image data to determine the concentration of the sample 165. Negative pressure, oil, and / or another substance can be used to push the reagent 178 and the portion of the sample 165 between the inlet 206 and the outlet 208. The first plate 200 may alternatively be hingedly coupled or removably coupled to the second plate 202 to allow the contact dispenser 145 to dispense the portion of the reagent 178 and sample 165 onto the second plate 202 before the first plate 200 is positioned on top of the second plate 202 (e.g., see Figure 4 ).

[0091] The drive assembly 173 includes a pump drive assembly 219 and a valve drive assembly 220. The controller 176 includes a user interface 221, a communication interface 222, one or more processors 224, and a memory 226, which stores instructions that can be executed by the one or more processors 224 to perform various functions including the disclosed specific implementation. The user interface 221, the communication interface 222, and the memory 226 are electrically and / or communicatively coupled to the one or more processors 224.

[0092] In a specific implementation, the user interface 221 receives input from the user and provides the user with information associated with the operation of the system 100 and / or the analysis performed. The user interface 221 may include a touch screen, a display, a keyboard, a speaker, a mouse, a trackball, and / or a voice recognition system. The touch screen and / or the display may display a graphical user interface (GUI).

[0093] In a specific implementation, the communication interface 222 uses a network to implement communication between the system 100 and a remote system (e.g., a computer). The network may include an intranet, a local area network (LAN), a wide area network (WAN), an intranet, etc. Some communications provided to the remote system may be associated with a quantification process generated or otherwise obtained by the system 100. Some communications provided to the system 100 may be associated with an amplification process, a purification process, a library normalization process, and / or a pooling process to be performed by the system 100.

[0094] The one or more processors 224 and / or the system 100 may include one or more of a processor-based system or a microprocessor-based system. In some implementations, the one or more processors 224 and / or the system 100 include a reduced instruction set computer (RISC), an application specific integrated circuit (ASIC), a field programmable gate array (FPGA), a field programmable logic device (FPLD), a logic circuit, and / or another logic-based device that performs various functions including those described herein.

[0095] Memory 226 may include one or more of a hard drive, flash memory, read-only memory (ROM), erasable programmable read-only memory (EPROM), electrically erasable programmable read-only memory (EEPROM), random access memory (RAM), nonvolatile RAM (NVRAM) memory, compact disk (CD), digital versatile disk (DVD), cache, and / or any other storage device or storage disk in which information is stored for any duration (e.g., permanently, temporarily, for an extended period of time, for buffering, for caching).

[0096] Figure 2 It can be used to implement Figure 1 162 of the substrate 400. The substrate 400 includes a pair of plates 200, 202 and spacers 402 positioned between the plates 200, 202 to define a gap 204 between the plates 200, 202. The spacers 402 are ball bearings 404 having opposing flat surfaces 406, 408. The substrate 400 may include three ball bearings 404 to define the gap 204 between the plates 200, 202 to be approximately 500 μm. As explained herein, the phrase "approximately 500 μm" means + / - 5 μm of 500 μm, including 500 μm itself and / or taking into account manufacturing tolerances.

[0097] Portions 410 of sample 165 and reagent 178 are shown positioned between plates 200, 202. Portions 410 may be referred to as droplets. Each of portions 410 may be associated with a different sample 165. Portions 410 may each be approximately 2 μL and have a nominal diameter of about 2 mm.

[0098] Figure 3 yes Figure 24. The cross-sectional view of the substrate 400 of FIG. 400 shows a channel 412 defined between the plates 200, 202 and a seal 414 defining the channel 412 between the plates 200, 202. The plates 200, 202 also define the channel 412. The seal 414 can be a flexible gasket seal. The channel 412 of the substrate 400 can be rinsed and / or cleaned with a liquid such as ethanol and purged with air after the quantification process is completed. For example, a portion 410 of the sample 165 and the reagent 178 can flow to the waste reservoir 214.

[0099] Figure 4 yes Figure 2 4 is a cross-sectional view of a substrate 400 showing the first plate 200 removed from the second plate 202 and the contact dispenser 145 dispensing a portion 410 onto the second plate 202.

[0100] Figure 5 yes Figure 2 4 is a cross-sectional view of substrate 400, which shows first plate 200 positioned on top of second plate 202 and imaging system 164 obtaining image data of one of portions 410. Imaging system 164 and / or system 100 may use the image data to determine an intensity value and a diameter of portion 410. Thereafter, imaging system 164 and / or system 100 may calculate a concentration of portion 410, for example, using a total fluorescence measurement.

[0101] The imaging system 164 may also or alternatively be used to perform a calibration process, wherein the imaging system 164 obtains image data of the portion 410, and the imaging system 164 and / or the system 100 calibrates the contact dispenser 145 by determining the volume of the portion 410, comparing the determined volume of the portion 410 to a reference volume, and determining a difference between the determined volume and the reference volume. If the difference between the determined volume and the reference volume is greater than a threshold value, the system 100 may calibrate and / or change the amount of liquid dispensed by the contact dispenser 145.

[0102] Figure 6 It can be used to implement Figure 1 162 is a top isometric view of a specific implementation of substrate 500 . Figure 6 The substrate 500 is similar to Figure 2 substrate 400. Figure 6The substrate 500 includes a mask 502, however, in the specific implementation shown, the mask is an opaque mask 504. The mask 502 defines a window 506 that allows the imaging system 164 to obtain image data of a portion 510 of the sample 165 and the reagent 178. The portion 510 can be a 2 μL droplet. The window 506 can be referred to as an unmasked area. An area 508 of the mask 502 can be relatively well defined and is shown surrounding the window 506. The imaging system 164 can alternatively be used to measure the area 508 of the mask 502. The imaging system 164 can be capable of measuring the area 508 of the mask 502 with an accuracy of about 5%. However, the measurement accuracy of the imaging system 164 may vary.

[0103] The substrate 500 may include spacers for accurately defining the gap 204 between the plates 200, 202. The gap 204 may be between about 200 μm and about 500 μm. The imaging system 164 may alternatively be used to measure the gap 204. The imaging system 164 may be capable of measuring the gap 204 with an accuracy of about 1%. The imaging system 164 may be capable of determining the liquid volume of the portion 510 with an accuracy of about 7%.

[0104] Figure 7 yes Figure 6 506 and 510. The imaging system 164 and / or system 100 may use the image data to determine an intensity value of the portion 510 that can be viewed through the window 506. The imaging system 164 and / or system 100 may use the intensity value and the thickness 512 of the gap 204 to determine the concentration of the portion 510. The channel 412 of the substrate 500 may be rinsed and / or cleaned with a liquid such as ethanol and purged with air after the quantification process is completed.

[0105] Figure 8 It can be used to implement Figure 1 162 is a cross-sectional view of a specific implementation of substrate 600 . Figure 8 The substrate 600 is similar to Figure 7 substrate 500. Figure 8The substrate 600 of the embodiment of the present invention includes a divider 602, however, the divider is coupled between the plates 200, 202 and defines a first channel 604 and a second channel 606. In other words, the substrate 600 has a plurality of channels 604, 606. Each of the channels 604, 606 may have a corresponding inlet 206 and / or a corresponding outlet 208. Each channel 604, 606 may therefore receive a different sample, and multiple samples may be processed and / or flow through the substrate 600 at the same or similar time. Although the substrate 600 is shown as having two channels 604, 606, the substrate 600 may have any number of channels 604, 606.

[0106] Fig. 9 It can be used to implement Figure 1 162 is a top isometric view of a specific implementation of substrate 700 . Fig. 9 The substrate 700 is similar to Figure 6 substrate 500. Fig. 9 The substrate 700 includes a first inlet 702, a second inlet 703, a third inlet 704 and an outlet 208 that are in fluid communication with the gap 204. Portions 705 of the sample 165 and the reagent 178 are shown as being separated by another fluid 706. Portions 705 may be referred to as droplets. Fluid 706 may be oil. The contact dispenser 145 deposits portion 705 in the second inlet 703 during operation, and fluid 706 is dispensed into the first inlet 702 and / or the third inlet 704. Fluid 706 separates portion 705. Channel 412 may be under negative pressure, and the negative pressure may sequentially push portion 705 between the second inlet 703 and the outlet 208. Fluid 706 may also or alternatively be used to push portion 705 between the second inlet 703 and the outlet 208.

[0107] Fig.10 yes Fig. 9 506 and 506. The imaging system 164 obtains image data of each of the portions 705 when the corresponding portion is aligned with the window 506. The imaging system 164 and / or the system 100 can use the image data to determine the intensity value of the portion 510 that can be viewed through the window 506. The imaging system 164 and / or the system 100 can use the intensity value and the thickness 512 of the gap 204 to determine the concentration of the portion 510. The channel 412 of the substrate 500 can be rinsed and / or cleaned using the fluid 706 and / or using a liquid such as ethanol after the quantification process is completed. The channel 412 of the substrate 700 can be dried by blowing the substrate 700 with air.

[0108] Fig.11 and Fig.12 Illustrated description for use Figures 1 to 10 Flowcharts of processes for substrate 162, 400, 500, 600, 700 and imaging system 164 of any of the other implementations disclosed herein. The order of execution of the blocks may be changed, and / or some of the blocks described may be changed, eliminated, combined, and / or subdivided into multiple blocks.

[0109] Fig.11 The process of the present invention begins by dispensing a sample 165 onto a first plate 202 of a substrate 400 (block 1102). The dispensed sample 165 may also be referred to as a portion of the sample 165. The second plate 200 of the substrate 400 is positioned on the first plate 202, and a plurality of spacers 402 are used to define a gap 204 between the first plate 202 and the second plate 200 (block 1104). In some implementations, the spacers 402 are ball bearings 404 having opposing flat surfaces 406, 408. The seal 414 is positioned around the perimeter of the first plate 202 and the second plate 204 and between the first plate 202 and the second plate 200 (block 1106). Image data of the sample 165 is obtained (block 1108), and the image data is used to determine the concentration of the sample 165 (block 1110). In some implementations, the imaging system 164 obtains the image data of the sample 165, and the imaging system 164 and / or the system 100 use the image data to determine the concentration of the sample 165.

[0110] Fig.12 The process of begins by dispensing a sample 165 in an inlet 206, 703 of a channel 212, 412 of a substrate 162, 700 (block 1202). The dispensed sample 165 may also be referred to as a portion of a sample 165. A fluid 706 is dispensed into the channel 212, 412 to separate the sample 165 from another sample 165 (block 1204). In some implementations, the fluid 706 includes oil. The fluid 706 may be dispensed into the channel 212 by dispensing the fluid into the second inlet 702 and / or the third inlet 704 of the channel 212. Image data of the sample 165 is obtained (block 1206). The image data may be obtained through a window 506 of the substrate 162, 700. The window 506 may be an unmasked area, and the masked area 502 may surround the unmasked area. The image data may be used to determine the concentration of the sample 165 (block 1208), and the sample 165 may flow out of the outlet 208 of the channel 212, 412 (block 1210). Flowing the sample 165 out of the outlet 208 of the channel 212, 412 may include using negative pressure and / or using the fluid 706 to push the sample 165 out of the outlet 208 of the channel 212, 412.

[0111] Fig.13 It can be used to implement Figure 11300 and imaging system 164 of substrate 162 and imaging system 164. Substrate 1300 may be implemented by a microfluidic card and / or may be made of a pair of plates. Substrate 1300 has a channel 1302 having an inlet 1304 and an outlet 1306, and a valve 1308 disposed between inlet 1304 and outlet 1306. Channel 1302 has a first portion 1310 located between valve 1308 and inlet 1304 and a second portion 1312 located between valve 1308 and outlet 1306. In operation, a portion of sample 165 is received in first portion 1310 of channel 1302 of substrate 1300, and imaging system 165 obtains image data of the portion of sample 165. The image data is used to determine the concentration of the portion of sample 165.

[0112] After obtaining image data of a portion of the sample 165, the portion of the sample 165 flows from the first portion 1310 through the valve 1308, the second portion 1312, and the outlet 1306. The valve 1308 may be implemented by a mechanical valve such as an elastomeric valve, a rotary face seal valve, etc. However, the valve 1308 may be implemented by other mechanical valves. For example, the valve 1308 may be implemented by a rotary valve, a pinch valve, a plate valve, a solenoid valve, a check valve, and / or a piezoelectric valve. If the valve 1308 is implemented by a mechanical valve, the valve 1308 may be actuated to an open position to allow the portion of the sample 165 to flow from the first portion 1310 through the valve 1308, the second portion 1312, and the outlet 1306. Alternatively, the valve 1308 may be implemented by a virtual valve, as further disclosed below.

[0113] Also included is a pump 1314 for moving the sample 165 from the inlet 1304 to the outlet 106. The pump 1314 may be used to achieve Figure 1 The pump 215 of FIG. 1314 has a plate 1316 including a rotation axis 1318 and an actuator 1320 for rotating the plate 1316. The base plate 1300 is coupled to the plate 1316 and extends radially from the rotation axis 1318.

[0114] A second substrate 1324 is shown coupled to plate 1316 and extends radially from rotation axis 1318. Second substrate 1324 may be the same or similar to substrate 1300. Plate 1316 has a first position 1326 and a second position 1328, and substrate 1300 is shown positioned at first position 1326 and second substrate 1324 is shown positioned at second position 1328. First position 1326 is opposite second position 1328. Second substrate 1324 may be a mirror image of substrate 1300 on plate 1316.

[0115] The pump 1314 includes a light access region 1330. The light access region 1330 allows the imaging system 164 to obtain image data of the sample 165 within the channel 1302. The actuator 1320 can rotate the base plate 1300 and / or the second base plate 1324 to align the imaging system 164 with the corresponding base plate 1300, 1324 to allow imaging data to be obtained.

[0116] The waste reservoir 214 is shown as being fluidly coupled to the outlet 1306. The waste spout 1331 is shown extending from the outlet 1306 into the waste reservoir 214. In the illustrated embodiment, the waste reservoir 214 includes an exhaust port 1332. The exhaust port 1332 may include a porous glass material 1334. For example, the porous glass material 1334 may be used to limit the escape of the atomized sample. The waste reservoir 214 may be removable for draining and / or cleaning. Therefore, the waste reservoir 214 may be reusable. As an example, the waste reservoir 214 may alternatively be a disposable microfluidic card assembly.

[0117] Fig.14 yes Fig.13 1300 and a top view of a substrate 1300 and a pump 1314. In the illustrated implementation, the valve 1308 is illustrated as a virtual valve 1336. The virtual valve 1136 is formed by a constriction 1338. The constriction 1338 reduces the width of the channel 1302. The first portion 1326 of the channel 1302 has a first width, the second portion 1328 of the channel 1302 has a second width, and the virtual valve 1336 has a third width. The third width is less than the first width and the second width. The virtual valve 1336 may be associated with a sharp expansion of the channel 1302 geometry after a sharp contraction of the channel 1302 geometry. The virtual valve 1336 may be patterned into the substrate 1300, or formed at the micro-to-macro junction between the substrate 1300 and / or the channel 1302 of the substrate 1300 and the waste spout 1331.

[0118] In some examples, channel 1302 may have a height between about 200 microns and about 500 microns. However, channel 1302 may have different heights. As an example, channel 1302 may hold about 2 microliters to about 200 microliters.

[0119] In the illustrated implementation, substrate 1300 includes mask 1340. Mask 1340 can be an opaque mask. Mask 1340 defines a window 1342 that allows imaging system 164 to obtain image data of a portion of sample 165, for example.

[0120] Fig.15 yes Fig.13Detailed view of a portion of the substrate 1300 and pump 1314. In the illustrated implementation, an inlet port 1344 is positioned at the inlet 1304, which funnels a portion of the sample 165 into the channel 1302. The inlet port 1344 includes a flow channel 1346. The flow channel 1346 is shown as a truncated cone-shaped flow channel 1348. However, the flow channel 1346 can have a different shape. As an example, the flow channel 1346 can be cylindrical.

[0121] The inlet port 1344 includes a first surface 1350 and a second surface 1352. The first surface 1350 is coupled to the base plate 1300. The frustoconical flow channel 1348 has a base opening 1354 located at the first surface 1350 and a top opening 1356 located at the second surface 1352. In some implementations, the base opening 1354 has a perimeter, and the inlet 1304 of the channel 1302 can be tangential to the base opening 1354. For example, the base opening 1354 can be radially offset from the inlet 1304.

[0122] Fig.16 It can be used to implement Figure 1 162 and a circular substrate 1400 of the pump 215. The circular substrate 1400 includes a plurality of channels 1302, each of which has an inlet 1304 and an outlet 1306, and a valve 1308 disposed between the inlet 1304 and the outlet 1306. The circular substrate 1400 can be implemented by a plastic disk and / or a circular wafer. The circular substrate 1400 includes a rotation axis 1318 and also includes an actuator that can be used to rotate the circular substrate 1400 around the rotation axis 1318. For example, the actuator can be a rotary actuator. In the illustrated implementation, the channels 1302 extend radially from the rotation axis 1318.

[0123] Figure 17 to Figure 18 A flowchart for implementing a method of the examples disclosed herein is illustrated. The order of execution of the blocks may be changed, and / or some of the blocks described may be changed, eliminated, combined and / or subdivided into multiple blocks.

[0124] Fig.17The process begins with the substrate 1300, 1400 rotating about the rotation axis 1318 at a first rate to draw a portion of the sample 165 into the first portion 1310 of the channel 1302 of the substrate 1300, 1400 (block 1502). The substrate 1300, 1400 includes the channel 1302, an inlet 1304 in fluid communication with the channel 1302, an outlet 1306, and a valve 1308 disposed between the inlet 1304 and the outlet 1306. The channel 1302 includes a first portion 1310 located between the valve 1308 and the inlet 1304 and a second portion 1312 located between the valve 1308 and the outlet 1306. The valve 1308 may include a virtual valve 1336. The virtual valve 1336 may include a constriction 1338.

[0125] The rotation of the substrate 1300, 1400 is stopped (block 1504), and image data of the portion of the sample 165 is obtained (block 1506). For example, the image data can be used to determine the concentration of the portion of the sample 165.

[0126] The substrate 1300, 1400 is rotated at a second rate about the rotation axis 1318 to push a portion of the sample 165 through the valve 1308 and into the second portion 1312 of the channel 1302 of the substrate 1300, 1400 and out of the outlet 1306 (block 1508). The portion of the sample 165 is received in a waste reservoir 214 fluidly coupled to the outlet 1306 (block 1510). The channel 1302 is flushed by flowing a flushing fluid through the channel 1302 and between the inlet 1304 and the outlet 1306 (block 1512).

[0127] Fig.18 The process begins with the substrate 1300, 1400 rotating about the rotation axis 1318 to draw a portion of the sample 165 into the first portion 1310 of the channel 1302 of the substrate 1300, 1400 (block 1602). The substrate 1300, 1400 includes the channel 1302, an inlet 1304 in fluid communication with the channel 1302, an outlet 1306, and a valve 1308 disposed between the inlet 1304 and the outlet 1306. The channel 1302 includes a first portion 1310 located between the valve 1308 and the inlet 1304 and a second portion 1312 located between the valve 1308 and the outlet 1306. The rotation of the substrate 1300, 1400 is stopped (block 1604).

[0128] Image data of a portion of the sample 165 is obtained (block 1606). The image data can be used to determine the concentration of the portion of the sample 165. The valve 1308 is actuated to an open position (block 1608), and the substrates 1300, 1400 are rotated about the rotation axis 1318 to push the portion of the sample 165 through the valve 1308 and into the second portion 1312 of the channel 1302 of the substrates 1300, 1400 and out of the outlet 1306 (block 1610). In this specific implementation, the valve 1308 can be implemented by a mechanical valve. The channel 1302 is flushed by flowing a flushing fluid through the channel 1302 and between the inlet 1304 and the outlet 1306 (block 1612). The flushing fluid can be removed from the channel 1302 by the centrifugal force of the rotating substrates 1300, 1400.

[0129] The above description is provided to enable those skilled in the art to practice the various configurations described herein. Although the subject technology has been described in detail with reference to various drawings and configurations, it should be understood that these drawings and configurations are only for illustrative purposes and should not be considered to limit the scope of the subject technology.

[0130] As used herein, an element or step recited in the singular and preceded by the word "one" or "a kind of" should be understood not to exclude a plurality of said elements or steps, unless such exclusion is expressly indicated. In addition, reference to "a specific implementation" is not intended to be interpreted as excluding the existence of additional specific implementations that also include the described features. In addition, unless expressly stated to the contrary, a specific implementation that "includes" or "has" one or more elements having a particular property may include additional elements, whether or not they have that property. In addition, the terms "includes," "has," etc. are used interchangeably herein.

[0131] The terms "substantially", "approximately" and "about" used throughout this specification are used to describe and illustrate small fluctuations, such as small fluctuations caused by changes in processing. For example, they can refer to less than or equal to ±5%, such as less than or equal to ±2%, such as less than or equal to ±1%, such as less than or equal to ±0.5%, such as less than or equal to ±0.2%, such as less than or equal to ±0.1%, such as less than or equal to ±0.05%. In one example, these terms include the absence of a variation of -0%.

[0132] There may be many other ways to realize the subject technology. Without departing from the scope of the subject technology, the various functions and elements described herein may be divided differently from those shown and elements. Various modifications to these specific implementations may be apparent to those skilled in the art, and the general principles defined herein may be applied to other specific implementations. Therefore, without departing from the scope of the subject technology, those of ordinary skill in the art may make many changes and modifications to the subject technology. For example, different numbers of given modules or units may be adopted, one or more different types of given modules or units may be adopted, given modules or units may be added or given modules or units may be omitted.

[0133] Underlined and / or italicized headings and subheadings are used for convenience only, do not limit the subject technology, and are not referenced in conjunction with the explanation of the description of the subject technology. All structural and functional equivalents of the elements of each specific implementation described throughout the present disclosure that are known or will later be known to those of ordinary skill in the art are expressly incorporated herein by reference and are intended to be covered by the subject technology. In addition, nothing disclosed herein is intended to be dedicated to the public, regardless of whether such disclosure is explicitly recited in the above description.

[0134] It should be understood that all combinations of the foregoing concepts and the additional concepts discussed in more detail below (assuming such concepts are not mutually inconsistent) are contemplated as being part of the subject matter disclosed herein. In particular, all combinations of the claimed subject matter appearing at the end of this disclosure are contemplated as being part of the subject matter disclosed herein.

Claims

1. A device, comprising: a substrate including a pair of plates and a plurality of spacers positioned between the plates to define a gap between the pair of plates; and Imaging system, A portion of a sample is to be received within the gap of the substrate, and the imaging system is to obtain image data of the portion of the sample, the image data being used to determine a concentration of the portion of the sample.

2. The device of claim 1, wherein the spacer comprises a ball bearing having opposing flat surfaces.

3. The device of any of the preceding claims, wherein the plurality of spacers comprises three spacers.

4. The device of any preceding claim, wherein the gap is about 500 microns.

5. A device according to any one of the preceding claims, wherein a channel is defined between the pair of plates.

6. The apparatus of claim 5, wherein the base plate further comprises a seal positioned between the pair of plates and defining the channel.

7. The apparatus of claim 6, wherein the seal is positioned around a perimeter of the pair of plates.

8. The apparatus of any preceding claim, wherein the substrate comprises a mask. The device of claim 8 , wherein the mask is an opaque mask.

10. An apparatus according to any preceding claim, wherein the substrate comprises an inlet and an outlet in fluid communication with the gap.

11. The device of any preceding claim, wherein the substrate comprises a plurality of channels.

12. The apparatus of any preceding claim, wherein the substrate comprises a first inlet, a second inlet, a third inlet and an outlet in fluid communication with the gap.

13. A method comprising: dispensing a sample onto a first plate of a substrate; positioning a second plate of the substrate on the first plate and defining a gap between the first plate and the second plate using a plurality of spacers; obtaining image data of the sample; as well as The image data is used to determine a concentration of the sample.

14. The method of claim 13, wherein the spacer comprises a ball bearing having opposing flat surfaces.

15. The method of any one of claims 13 to 14, wherein the plurality of spacers comprises three spacers.

16. The method according to any one of claims 13 to 15, further comprising: A seal is positioned around a perimeter of the first plate and the second plate.

17. A method comprising: dispensing a sample into an inlet of a channel of a substrate; obtaining image data of the sample; using the image data to determine a concentration of the sample; as well as The sample is allowed to flow out of the outlet of the channel.

18. The method of claim 17, wherein obtaining the image data comprises obtaining the image data through a window of the substrate.

19. The method of claim 18, wherein the window comprises an unmasked area, and wherein a masked area can surround the unmasked area.

20. The method according to any one of claims 17 to 19, further comprising: A fluid is dispensed into the channel to separate the sample from another sample.

21. The method of claim 20, wherein the fluid comprises oil.

22. The method of any one of claims 20 to 21, wherein dispensing the fluid into the channel comprises dispensing the fluid into a second inlet of the channel.

23. The method of any one of claims 17 to 22, wherein causing the sample to flow out of the outlet of the channel comprises using negative pressure to push the sample out of the outlet of the channel.

24. The method of any one of claims 17 to 22, wherein causing the sample to flow out of the outlet of the channel comprises using a fluid to push the sample out of the outlet of the channel.

25. An apparatus, comprising: A substrate, comprising: a channel, the channel comprising an inlet and an outlet; and a valve disposed between the inlet and the outlet, wherein the passage includes a first portion between the valve and the inlet and a second portion between the valve and the outlet; and Imaging system, A portion of a sample is to be received within the first portion of the channel of the substrate, and the imaging system is to obtain image data of the portion of the sample, the image data being used to determine a concentration of the portion of the sample.

26. The device of claim 25, further comprising a pump for moving the sample from the inlet to the outlet.

27. The device of claim 26, wherein the pump comprises a plate including a rotational axis and an actuator for rotating the plate, the base plate being coupled to the plate and extending radially from the rotational axis.

28. The apparatus of claim 27, further comprising a second substrate coupled to the plate and extending radially from the rotation axis.

29. The apparatus of claim 28, wherein the plate includes a first position and a second position, the substrate being positioned at the first position and the second substrate being positioned at the second position, the first position being opposite to the second position.

30. The apparatus of any one of claims 28 to 29, wherein the second substrate comprises a mirror image of the substrate on the plate.

31. The device of any one of claims 26 to 31, wherein the pump comprises a light access region.

32. The device of any one of claims 25 to 31 further comprising a waste reservoir fluidly coupled to the outlet.

33. The apparatus of claim 32, wherein the waste reservoir comprises a vent.

34. The apparatus of claim 33, wherein the exhaust port comprises a porous glass frit.

35. The device of claim 32, wherein the waste reservoir comprises a disposable microfluidic card assembly.

36. An apparatus according to any one of claims 25 to 32, wherein the valve comprises a virtual valve.

37. The device of claim 36, wherein the virtual valve comprises a constriction.

38. An apparatus, comprising: A substrate, comprising: a channel, the channel comprising an inlet and an outlet; and a valve disposed between the inlet and the outlet, wherein the passage comprises a first portion between the valve and the inlet and a second portion between the valve and the outlet, A portion of a sample is to be received within the first portion of the channel of the substrate, and an imaging system is to obtain image data of the portion of the sample, the image data being used to determine a concentration of the portion of the sample.

39. The apparatus of claim 38, wherein after obtaining the image data of the portion of the sample, the portion of the sample is to flow from the first portion through the valve, the second portion, and the outlet.

40. The device of any one of claims 38 to 39, wherein the valve comprises a virtual valve.

41. The device of claim 40, wherein the virtual valve comprises a constriction.

42. The device of any one of claims 38 to 41, wherein the first portion of the channel has a first width, the second portion of the channel has a second width, and the virtual valve has a third value, the third width being less than the first width and the second width.

43. The device of any one of claims 38 to 42, wherein the channel comprises a height between about 200 microns and about 500 microns.

44. An apparatus according to any one of claims 38 to 43, wherein the substrate comprises a mask.

45. The device of claim 44, wherein the mask is an opaque mask.

46. ​​A device according to any one of claims 38 to 45, wherein the substrate comprises a microfluidic card.

47. The device of any one of claims 38, 39, 43 to 46, wherein the valve comprises an elastomeric valve or a rotating face seal valve.

48. The device of any one of claims 38 to 47, further comprising an inlet port at the inlet for funneling the portion of the sample into the channel.

49. The apparatus of claim 48, wherein the inlet port comprises a flow passage, the flow passage comprising a frusto-conical flow passage.

50. A device according to claim 49, wherein the inlet port includes a first surface and a second surface, the first surface is connected to the substrate, the truncated conical flow channel includes a base opening located at the first surface and a top opening located at the second surface, the base opening has a periphery, and the inlet is tangent to the base opening.

51. An apparatus, comprising: A circular substate, the circular substate comprising a plurality of channels, each channel comprising: Entrance; export; and A valve is disposed between the inlet and the outlet, wherein the passage includes a first portion between the valve and the inlet and a second portion between the valve and the outlet.

52. The apparatus of claim 51, wherein the circular substrate comprises a plastic disk.

53. The device of claim 51, wherein the circular substrate comprises a circular wafer.

54. The device according to any one of claims 51 to 53, wherein the circular substrate includes a rotation axis, the circular substrate further comprising an actuator for rotating the circular substrate about the rotation axis, and the channel extends radially from the rotation axis.

55. A method comprising: Rotating a substrate about a rotation axis at a first rate to draw a portion of a sample into a first portion of a channel of the substrate, the substrate comprising: the channel; an inlet; an outlet, the outlet being in fluid communication with the channel; and a valve disposed between the inlet and the outlet, wherein the channel comprises the first portion between the valve and the inlet and a second portion between the valve and the outlet; stopping the rotation of the substrate; obtaining image data of the portion of the sample, the image data to be used to determine a concentration of the portion of the sample; The substrate is rotated about the rotation axis at a second rate to push the portion of the sample through the valve and into the second portion of the channel of the substrate and out of the outlet.

56. The method of claim 55, further comprising: The portion of the sample is received in a waste reservoir fluidly coupled to the outlet.

57. The method according to any one of claims 55 to 56, further comprising: The channel is flushed by flowing a flushing fluid through the channel and between the inlet and the outlet.

58. A method according to any one of claims 55 to 57, wherein the valve comprises a virtual valve.

59. The method of claim 58, wherein the virtual valve comprises a constriction.

60. A method comprising: Rotating a substrate about a rotation axis to draw a portion of the sample into a first portion of a channel of the substrate, the substrate comprising: the channel; an inlet; an outlet, the outlet being in fluid communication with the channel; and a valve disposed between the inlet and the outlet, wherein the channel comprises the first portion between the valve and the inlet and a second portion between the valve and the outlet; stopping the rotation of the substrate; obtaining image data of the portion of the sample, the image data to be used to determine a concentration of the portion of the sample; actuating the valve to an open position; and The substrate is rotated about the rotation axis to push the portion of the sample through the valve and into the second portion of the channel of the substrate and out of the outlet.

61. The method of claim 60, further comprising: The channel is flushed by flowing a flushing fluid through the channel and between the inlet and the outlet.

Images