Apparatus and method for measuring parameters of particles - Patents.com

Abstract

A portable device for measuring at least one parameter of particles in a solution, comprising a light source, a camera with an imaging sensor for generating a digital image, an optical system comprising at least one objective lens and / or a combination of objective lenses and / or lenses and a sample holder, wherein the sample comprises particles in solution, and wherein the imaging sensor and / or the camera have a frame rate greater than 100 frames / sec, preferably greater than 200 frames / sec, more preferably greater than 290 frames / sec and even more preferably greater than 300 frames / sec.

Description

[Technical field]

[0001] The present invention relates to an apparatus and method for measuring at least one parameter of particles, in particular particles in a solution. [Background technology]

[0002] Dairy farmers are losing money every year due to low conception rates in their cows, which have declined by 20% over the past 40 years. Current semen evaluation methods used on farms have an error rate of more than 10% for standard semen concentrations and rely primarily on visual evaluation. There are no user-independent quality control standards industry-wide for veterinarians, farmers and artificial insemination technicians to check semen quality prior to breeding.

[0003] It is desirable to find methods to help dairy farmers increase conception rates to improve the profitability of their farm operations while reducing the carbon footprint of dairy and meat products. It is also desirable to find improved semen evaluation methods for humans (e.g., human fertility studies in IVF clinics) and other animals (including sheep, horses, pigs, goats, fish, poultry, dogs and all cattle). Summary of the Invention [Means for solving the problem]

[0004] According to a first aspect of the invention, there is provided a portable device for measuring at least one parameter of particles in a solution, comprising a light source, an imaging means (e.g. an imaging sensor) for generating an image (e.g. a digital image), an optical system comprising at least one objective lens and / or a combination of objective lenses and / or lenses and / or a sample holder.

[0005] According to a second aspect of the invention, there is provided a system comprising a portable device for measuring at least one parameter of particles in a solution and a remote processing unit, wherein the portable device comprises a light source, an imaging means (e.g. an imaging sensor) for generating an image (e.g. a digital image), an optical system comprising at least one objective lens, a sample holder and means for transferring the image (e.g. a digital image) to the remote processing unit.

[0006] The device may comprise a processing unit for and / or configured to process digital images. The remote processing unit may be for and / or configured to process digital images.

[0007] The processing unit and / or remote processing unit may comprise a central processing unit (CPU) and a graphics processing unit (GPU). The processing unit and / or remote processing unit may provide processing resources for automatically or semi-automatically processing digital images. The processing unit and / or remote processing unit may comprise a single circuit (e.g., a suitable processing circuit) or multiple circuits. In an embodiment, the one or more circuits may be implemented in the CPU and / or GPU, respectively, by a computer program having computer-readable instructions executable to perform the method of the embodiment. In other embodiments, the one or more circuits may be implemented as one or more ASICs (Application Specific Integrated Circuits) or FPGAs (Field Programmable Gate Arrays) or other suitable dedicated circuits. The computing device may comprise a processing unit and / or remote processing unit. The computing device, processing unit and / or remote processing unit may also include a hard drive and other components of a PC, including RAM, ROM, a data bus, an operating system including various device drivers, and hardware devices including a graphics card. While specific circuits may be used, in embodiments, one or more of the functions of those circuits may be provided by a single processing resource or other component, or the functions provided by a single circuit may be provided by a combination of two or more processing resources or other components. A reference to a single circuit encompasses multiple components that provide the functions of that circuit, whether or not such components are separate from one another, and a reference to multiple circuits encompasses a single component that provides the functions of those circuits. An embodiment or features thereof may be implemented as a computer program product for use with a computer system, the computer program product being a series of computer instructions stored on a data recording medium, such as, for example, a disk, CD-ROM, ROM, etc., or embodied in a computer data signal, the signal being transmitted via a tangible medium or a wireless medium, such as, for example, microwave or infrared.The series of computer instructions may constitute all or part of the above-mentioned functions, and may be stored in any volatile or non-volatile memory device, such as a semiconductor device, a magnetic device, an optical device, or other memory device. In addition, although the embodiments implement certain functions by software, those skilled in the art will appreciate that the functions may be implemented by hardware alone or by a combination of hardware and software. Thus, the embodiments are not limited to being implemented by software.

[0008] The processing unit and / or the remote processing unit may be for processing, configured to process, and / or comprise at least one pre-stored processing routine. The pre-stored processing routine may be for obtaining at least one parameter. The pre-stored processing routine may be configurable by a user. The at least one processing routine may include analyzing a power spectrum of the difference between pairs of spatial Fourier transforms of digital images separated by a time delay over a range of time delays and spatial Fourier frequencies. This is the difference between the two images in a pair (not the difference between pairs), which is then calculated for all pairs or a subsection of all pairs of Fourier images. The time delay is the time difference between the two images in a pair, not the time difference between pairs.

[0009] The acquisition of the at least one parameter may be performed by performing differential dynamic microscopy (DDM). The processing unit and / or the remote processing unit may be for and / or configured to perform differential dynamic microscopy (DDM). DDM may be performed on all image pairs and q values ​​or a subsection of all image pairs and / or a subsection of all q values.

[0010] The at least one parameter may be obtained by processing the digital image. The at least one parameter may be obtained by analyzing (and / or calculating) a Fourier transform of the digital image(s). The at least one parameter may be obtained by analyzing (and / or calculating) a spectrum (e.g., a power spectrum). The at least one parameter may be obtained by analyzing (and / or calculating) a difference spectrum (e.g., a power spectrum) between multiple (e.g., pairs) of (e.g., spatial) Fourier transforms of the digital image(s). The digital images may be separated by time delays, for example, over a range of time delays and / or spatial Fourier frequencies.

[0011] For example, analyzing the difference (e.g., power) spectrum between pairs of (e.g., spatial) Fourier transforms of digital images separated by a time delay over a range of time delays and / or spatial Fourier frequencies can include (1) calculating a differential image correlation function (DICF), e.g., calculating a spatial Fourier transform of the images, and then calculating a power spectrum of the difference between pairs of Fourier images, e.g., over a range of available delay times τ (i.e., time differences between two selected images) and / or spatial frequencies (q) provided by the Fourier transform of the images. All possible Fourier image pairs or selected Fourier image pairs can be considered. The calculation can be performed for all possible Fourier image pairs or a subsection of the Fourier image pairs. The method can further include (2) then averaging all resulting DICFs that have the same delay time (τ) together. The method can further include (3) then performing radial averaging for each q value, e.g., generating a final time-averaged and vectorially {q}-averaged DICF, e.g., as a function of delay time τ and / or spatial frequency q. All possible q values ​​or selected q values ​​can be considered. The calculations can be performed for all possible q values ​​or a subsection of the q values.

[0012] The optical system and imaging sensor may be configurable or may be configured such that the pixel size of the image is and / or selectable from the range of 0.1 microns / pixel to 10.0 microns / pixel, preferably 0.5 microns / pixel to 7 microns / pixel, more preferably 2 to 7 microns / pixel, 3 to 6 microns / pixel, 4 to 5 microns / pixel, 4 to 4.5 microns / pixel, 4.10 to 4.30 microns / pixel, 4.15 to 4.25 microns / pixel, 4.2 to 4.35 microns / pixel, 4.25 to 4.35 microns / pixel, greater than 2.65 microns / pixel and less than 7 microns / pixel, greater than 2.65 microns / pixel and less than 7.04 microns / pixel, greater than 2 microns / pixel and less than 2.65 microns / pixel. The pixel size of the image may be 2 microns / pixel, 4.2 microns / pixel, 4.3 microns / pixel, or 7 microns / pixel. It will be understood that image pixel sizes are considered to be substantially and / or approximately the values ​​indicated. Image pixel size values ​​may not necessarily be exactly as stated. For example, they may be within + / - 5% of the values. This may be due, for example, to tolerances in design for manufacture and / or variations in manufacturing.

[0013] The optical system and the image sensor have a q value of 0.05 μm for DDM. -1 to 0.4 μm -1 , or any subset of this range adjusted for the species, e.g., 0.2 μm for bull sperm. -1 to 0.4 μm -1 Other parameters can be extracted from different ranges of q values. The Q value can depend on both the pixel size of the image and the size of the image in pixels.

[0014] The device and / or system may include a camera. The camera may include an imaging sensor. The imaging sensor and / or the camera may capture digital images and / or video.

[0015] The pixel size of the image may depend on several factors, such as the pixel size of the imaging sensor, the setup of the optical system (e.g., optical components such as an objective lens or other lenses, and the positions between the optical components in the optical system including the imaging sensor), and how the imaging sensor (or more specifically, the camera including the imaging sensor) is operating, e.g., the binning mode used or set.

[0016] The device and / or system may comprise means for displaying at least one parameter of the particles in the solution. The means for displaying at least one parameter of the particles in the solution may be a screen or a printer for generating a paper report.

[0017] The device and / or system may include a means for user interaction (such as a button) for (e.g., automatically) performing data collection and / or data analysis. The data collection and / or data analysis may be automated to generate measurements of parameters of the particles. The data collection and / or data analysis may be performed by a single user interaction such as the click of a button (or, more generally, the means for user interaction). Data collection and / or data analysis of measurements (such as a series of measurements) over time (e.g., over several hours, such as up to 12 hours) may be automatically collected by a single user interaction such as the click of a button.

[0018] The device and / or system may be pre-configured to provide a desired image pixel size, i.e., one or more of the factors that result in a particular image pixel size may be arranged or set such that the image pixel size is set to have a predetermined range or value.

[0019] The at least one objective lens may comprise an objective lens, a lens, or a combination of lenses.

[0020] At least one objective lens and / or device and / or other components of the optical system may be associated with one or more processing routines.

[0021] The at least one objective lens may have at least one optical characteristic. The at least one optical characteristic may be a magnification, a field of view, a depth of field, a focal length. The at least one optical characteristic may be related to providing a desired image pixel size.

[0022] The optical system may comprise at least two objective lenses. The at least two objective lenses may have at least one different optical property. The at least two objective lenses may each have a different magnification. One of the objective lenses may be for enabling an image to be captured for DDM processing (e.g., for measuring the motility of microorganisms). The other of the objective lenses may be for visual inspection of particles (e.g., for visualizing particles in solution at a resolution at which the heads of microorganisms are visible). As an example, this may be in the range of 5 to 15 pixels in size of the image, although this depends on the pixel size of the final image and therefore on the objective lens or combination of lenses in combination with the way the camera works (e.g., 1x1 or 2x2 binning). The other of the objective lenses may be used, for example, for smaller microorganisms (e.g., bacteria) and / or colloidal particles.

[0023] The objective lens may have a magnification such that the size of the head of the microorganism in the image is in the range of 1 to 5 pixels or less than 1 pixel.

[0024] The objective lens may have a focal length of 15 cm. The objective lens may have a focal length of less than 15 cm. This allows the device to be more compact.

[0025] The objective lens may have a magnification in the range of 1x to 4x (e.g., for sperm), 5x to 10x (e.g., for bacteria such as E. coli), 5x to 20x (e.g., for microalgae), and / or 5x to 50x for colloids.

[0026] The optical system may comprise a means for reflecting light from the objective lens to the imaging sensor. The optical system may comprise a mirror. The mirror may be between the at least one objective lens and the imaging sensor. The mirror may have the advantage that the components of the device fit into a stable and conveniently sized housing or box. In the absence of a mirror, all light from the light source to the imaging sensor may travel on a single straight line. By having a mirror that splits the distance from the light source to the imaging sensor into two non-straight parts, a more compact device may be realized. At least one objective lens may require a distance of about 30 cm between the light source and the imaging sensor. The mirror may split that distance into, for example, 2 x 15 cm parts.

[0027] The optical system may comprise means for refracting and / or focusing light from the objective lens to the imaging sensor. The optical system may comprise a further lens between the at least one objective lens and the imaging sensor. More specifically, the further lens may be between the mirror and the imaging sensor. The further lens may be called an adjustment lens. The further lens may allow fine tuning of the pixel size of the final image and / or parfocal imaging. Parfocal imaging means that (approximately) the same part of the sample is imaged when switching from two different objective lenses.

[0028] The imaging sensor may have a pixel size in the range of 0.5 to 10 microns / pixel, more preferably in the range of 2 to 7 microns / pixel, even more preferably in the range of 2 to 5 microns / pixel, 3 to 6 microns / pixel, 4 to 5 microns / pixel. The imaging sensor may have a pixel size of 4.8 microns. It will be understood that this pixel size of the imaging sensor is different from the pixel size of the image. For example, if the overall optical setup provides a magnification of 10 times, then the pixel size of the image = 0.1 x the pixel size of the imaging sensor. The values ​​of the pixel sizes in the imaging sensor may not necessarily be as stated. For example, they may be within a range of + / - 5% of the value.

[0029] The device and / or system may include means for operating in different binning and / or skipping modes. The imaging sensor and / or camera may be configured to operate in different binning and / or skipping modes, e.g., 1x1, 2x2, and 4x4 binning modes. This allows the pixel range of the desired image to be set so that, for example, one objective lens can be used to perform DDM on the sperm, and the other objective lens can be used to perform visual imaging of the sperm. The camera may have a driver that allows binning. Binning can be done in the camera or in post-processing after the image is captured and / or the video is recorded. "Binning" may be considered to allow pixel combinations to become one pixel, and "skipping" may be considered to skip pixels to reduce image resolution.

[0030] The imaging sensor and / or camera may have a frame rate of greater than 50 frames / sec, preferably greater than 100 frames / sec, more preferably greater than 200 frames / sec, and even more preferably greater than 300 frames / sec. In an embodiment, the imaging sensor and / or camera may have a frame rate of greater than 290 frames / sec. In an embodiment, the imaging sensor and / or camera may have a frame rate of at least one of up to 500, 600, 700, 800, 900, 1000, 10,000, 100,000, 1 million frames / sec. As an example, the imaging sensor and / or camera may have a range of frame rates from 300 to 800 frames / sec. It will be understood that this is merely an example and that there may be ranges including any applicable lower and upper limits described. More specifically, an imaging sensor having a frame rate is considered to mean that the electronics surrounding the imaging sensor have or set the frame rate. The camera is considered to have or set the frame rate. The frame rate may be a desired and / or predetermined frame rate. The digital images and / or videos may be recorded (e.g., by a processing unit or a remote processing unit) at a (selected) frame rate. The processing unit or the remote processing unit may be for or configured to process and / or record the digital images and / or videos at a desired predetermined frame rate. The camera may operate at a desired predetermined frame rate and the video may be recorded at the desired predetermined frame rate. There may be a variation in the exact frame rate, for example, perhaps within 1%. For example, if the desired frame rate is 300 fps, the final and true frame rate of the recorded images may vary between about 297 fps and 303 fps throughout the video. For example, a frame rate of 300 fps is considered most preferred (i.e., ideal), because other lower frame rates (such as the use of a frame rate of 200 fps) may struggle to apply the technique to all species or a wide range of specimens. However, it will be understood that other frame rates, for example 290 fps, are effective.The frame rates mentioned above are considered to be substantial or approximate frame rates. The frame rate that the imaging sensor and / or camera can transmit may depend on the size of the image (in pixels). For example, frame rates of 300 frames per second or greater may be achieved with image sizes of, for example, 512x512 pixels or 328x328 pixels. It will be understood that these image sizes are merely examples and other different image sizes may be used for a particular frame rate. Frame rates greater than 300 frames per second may have a field of view (image size) of at least 300x300 pixels (+ / -5%), or more preferably at least 328x328 pixels (+ / -5%). Frame rates greater than 100 frames per second may have a field of view (image size) of at least 512x512 pixels (+ / -5%). Frame rates greater than 200 frames per second may have a field of view (image size) of at least 300x300 pixels (+ / -5%), more preferably at least 328x328 pixels (+ / -5%). A frame rate greater than 290 frames per second may be a field of view (image size) of at least 300x300 pixels (+ / -5%), more preferably at least 328x328 pixels (+ / -5%). For example, an imaging sensor and / or camera may perform up to 500 frames per second with a field of view (image size) of at least 512x512 pixels (+ / -5%). As another example, an imaging sensor and / or camera may perform up to 800 frames per second with a field of view (image size) of at least 328x328 pixels (+ / -5%). It will be understood that other imaging sensors and / or cameras may have higher frame rates than these for the same or different image sizes, and that, at least in theory, there is no upper limit to the frame rate. Generally, cameras with relatively high frame rates are more expensive than cameras with relatively low frame rates. As an example, when recording for 30 seconds at 600 fps, technically there is twice the amount of images compared to 300 fps. Therefore, large video sizes and many images to process are required, and therefore long processing times and computer RAM.However, although it can record 600fps video, only the equivalent of a 300fps image can be used for processing, which helps solve the problem of long processing times.

[0031] The pixel size of an image may be the size of the portion of the sample that is imaged per pixel. The pixel size of an image represents what the pixel "sees" in the true sample. For example, if there is a 10 micron feature in the sample and the pixel size of the image is 1 micron, then this feature will be displayed over approximately 10 pixels in the image. The pixel size of an image may be considered to be the resolution (i.e., the shortest distance between two points on the sample that can be distinguished as a separate entity or individual unit by the device and / or system). A pixel is considered to be a discrete unit. The pixel size of an image may be referred to as the size of the portion of the sample that is imaged per discrete unit that can be distinguished, processed, or displayed.

[0032] The image pixel size may be greater than 2.65 microns / pixel and / or less than 7.04 microns / pixel (e.g., for sperm). Having an image pixel size of 2.65 microns / pixel, 7.04 microns / pixel, and / or between 2.65 microns / pixel and 7.04 microns / pixel may be effective for measuring at least one parameter of sperm, for example, but not all image pixel sizes within that range are optimal. There may be an optimal image pixel size and / or pixel size range between 2.65 microns / pixel and 7.04 microns / pixel. The device and / or system may be configured and / or configurable to provide an optimal image pixel size and / or pixel size range for the particular particle desired to be measured. The device and / or system is configurable and / or can be configured such that the image pixel size and / or range of pixel sizes provide a balance between having a wide enough field of view for statistical analysis and a range of q values ​​for efficiency of DDM analysis, and / or the magnification is not so large that visual imaging of the particles is more difficult or not possible when using another objective lens (i.e., the magnification is such that visual imaging of the particles is possible and not too difficult when using another objective lens) (these image pixel sizes and / or range of pixel sizes are believed to be optimal).

[0033] The image pixel size may be substantially and / or approximately 4.3 microns / pixel. It is believed that the image pixel size is 4.3+ / -5% microns / pixel. This may have the advantage of providing a balance between having a wide enough field of view for good statistics and a suitable q-value range for optimal efficiency of DDM analysis. 2.65 microns / pixel may give a lower suitable q-value for the technique to work, resulting in a 4x smaller field of view compared to 4.3 microns / pixel when using the same number of image pixels. This may potentially result in less accurate measurements and may require a relatively higher number of pixels to obtain a similar field of view, which may result in problems processing videos efficiently on low spec laptops, for example. For microorganisms larger than sperm, 2.65 microns / pixel may still work, but may not be optimal for the same reasons as above. 7.04 microns / pixel also means a significantly higher magnification compared to using 4.3 microns / pixel, and therefore observation of individual sperm cells and flagella may not be possible.

[0034] The pixel size of the image may be substantially and / or approximately 0.9 microns / pixel, 1.7 microns / pixel, 2.1 microns / pixel and / or 4.3 microns / pixel. 0.9 microns / pixel, or more specifically 0.86 microns / pixel, allows imaging of sperm and / or microalgae flagella (e.g., approximately 1 micron thick). 4.3 microns / pixel allows performing DDM on sperm and / or microalgae. 0.9 microns / pixel, 1.7 microns / pixel and / or 2.1 microns / pixel allows performing DDM on bacteria. 0.9 microns / pixel allows performing DDM on colloidal particles. The values ​​of the pixel size of the image are considered to be within + / - 5% of the value. These exemplary image pixel sizes may provide a balance between a sufficiently wide field of view for good statistics and an appropriate range of q values ​​for optimal efficiency of the DDM analysis.

[0035] At least one parameter of a particle is considered to be a parameter that characterizes the particle.

[0036] The device and / or system may be for measuring multiple parameters of particles in solution and / or parameters characterizing particles in solution.

[0037] The at least one parameter may include the mobility of the particle, preferably the rate of movement, the average velocity, the concentration, the size, the amplitude of the head movement, the diffusion velocity and / or the frequency of the head movement. More generally, the intensity variations in the image may be characterized. The processing unit and / or the remote processing unit may be for and / or configured to characterize the intensity variations in the image (e.g., using DDM). The imaging sensor and / or the camera is for detecting the intensity variations of the light transmitted through the particle in the solution. Characterizing the intensity variations in the image may consist of collecting the intensity variations (e.g., by detecting the intensity variations using the imaging sensor and / or the camera) and processing the image (e.g., using DDM) to obtain measurements.

[0038] The particle may be a microorganism.

[0039] Microorganisms can vary in size from bacteria to microalgae.

[0040] The microorganisms may be bacteria, sperm, and microalgae, with bacteria having a size of about 0.1 microns to 5 microns, sperm having a size of about 1 micron to 10 microns, and microalgae having a size of about 2 microns to 15 microns.

[0041] The particles may be colloids or colloidal particles. They may have a size ranging from 0.05 microns to 3 microns, more preferably from 0.1 microns to 3 microns, more commonly from tens of nanometers to micrometers. The particles may be droplets (e.g., milk fat droplets) and / or emulsions.

[0042] The sample may include particles in a solution. The sample holder may be for holding the sample (e.g., on a sample slide).

[0043] The device can be configured and / or configured such that light from the light source is transmitted to the sample holder without undergoing refraction. The device may not include a focusing lens. The device can be configured such that light from the light source is incident on the sample in the sample holder without refraction. The light can be directly incident on the sample in the sample holder. This has the advantage that the device can be physically smaller and lighter (i.e., easier to carry or move). Standard microscopes include a focusing lens to focus the light on the objective lens. This is because the microscope is intended to use high magnifications that require more light. A focusing lens is not required for the present device because it is based on low magnification and therefore does not require high illumination. Avoiding the use of a focusing lens does not adversely affect imaging and analysis, but allows for a reduction in the overall height of the device.

[0044] The light source may be an LED. The LED may be disposed in an LED holder. The LED may be of any color. The LED may preferably be green. This may be for bright field imaging of the sample.

[0045] The device may be at least partially enclosed in a housing (e.g., a box). The device and / or box may have dimensions of less than 40x30x20cm. The device and / or box may weigh less than 2.5kg. The device and / or box may weigh approximately 4kg. The device may comprise a housing.

[0046] The device and / or system may comprise a means for moving at least one objective lens (e.g., to a desired position). The device and / or system may comprise a means for switching between at least two objective lenses. The device and / or system may comprise an objective lens movement stage (e.g., a slider) for sliding (e.g., laterally) to switch between at least two objective lenses. That is, the slider may be configured to slide laterally to move between the two objective lenses such that light transmitted from the sample passes through one of the objective lenses or the other of the objective lenses. The means for moving at least one objective lens and / or the means for switching between at least two objective lenses sliders and / or the sliders may be configured to be moved manually (e.g., by a user moving them by hand) and / or controlled and / or automated (e.g., electronically or by a processing unit).

[0047] The device and / or system may comprise means for returning the at least one objective lens and / or the two objective lenses to the correct and / or exact position.

[0048] The device and / or system may comprise means for providing automatic detection of the position of the objective translation stage and / or the objective mounting plate.

[0049] The device and / or system may comprise means for heating, cooling or maintaining the temperature of the sample and / or the sample slide and / or the plurality of grooves of the sample slide and / or the sample holder and / or the sample stage. This may comprise an integrated sample heating and / or cooling system. The device may comprise a heating stage configured to heat and / or maintain the temperature of the sample (and / or the sample slide for holding the sample) at a predetermined temperature. The heating stage may be configured to heat and / or maintain the temperature of the groove or the plurality (e.g., four) of grooves of the sample slide at a predetermined temperature. Each or all of the plurality of grooves of the sample slide may be maintained at substantially the same predetermined temperature. The predetermined temperature may be fixed by the device (or system) as an inherent value of the device (or system) or may be selected / adjusted by a user, for example, through the use of software. The sample holder may hold the sample slide. The predetermined temperature may be between ambient temperature (e.g., 15 to 25° C.) and 50° C. The specimen and / or specimen slide may be maintained at a desired temperature, e.g., a temperature associated with animal reproduction, e.g., in the range of 36-41°C (which may, for example, be + / -0.1°C, + / -0.5°C or + / -1°C), e.g., substantially 37.5°C+ / -0.5°C (e.g., for bacteria or sperm), 36°C, 36°C+ / -0.1°C, 36°C+ / -0.5°C, 36°C+ / -1°C, 37°C, 37°C+ / -0.1°C, 37°C+ / -0.5°C, 37°C+ / -1°C, 37.5°C, The sample may be heated and / or maintained at 37.5° C.+ / -0.1° C., 37.5° C.+ / -0.5° C., 37.5° C.+ / -1° C., 38° C., 38° C.+ / -0.1° C., 38° C.+ / -0.5° C., 38° C.+ / -1° C., 39° C., 39° C.+ / -0.1° C., 39° C.+ / -0.5° C., 39° C.+ / -1° C., 40° C., 40° C.+ / -0.1° C., 40° C.+ / -0.5° C., 40° C.+ / -1° C., 41° C., 41° C.+ / -0.1° C., 41° C.+ / -0.5° C., and / or 41° C.+ / -1° C. Samples may be maintained at other temperatures depending on the particles, microorganisms, or colloids.

[0050] The heating stage may comprise a plate that is heated to a predetermined temperature. The heating stage may comprise at least one resistor or a plurality of resistors that heat the plate. The heating stage may comprise two resistors that heat the plate. The heating stage may comprise a temperature sensor that measures the temperature of the plate. The temperature may be read continuously during use and the temperature measurement may be available in real time. The resistor, the two resistors and / or the temperature sensor may be located in one or more holes in the heating stage. The device and / or system comprises a means for maintaining contact between the heating stage (e.g., plate) and the sample slide. The sample holder and heating stage are configured such that the sample slide is in contact with the heating stage (e.g., plate). This may be done by the plate lifting the slide.

[0051] The device and / or system may include a cooling system and / or cooling stage (e.g., to cool and / or maintain the temperature of the sample at a predetermined temperature). The cooling system or cooling stage may include a Peltier module, which allows for more precise control of temperature over a wider temperature range.

[0052] The device may comprise a heating stage and / or a cooling stage. The heating stage and the cooling stage may be combined into a heating / cooling stage.

[0053] The sample slide may include a groove or grooves for holding a sample. The sample holder may be configured to hold a sample slide with multiple grooves. The device and / or sample holder may be configured such that a predetermined position or positions (e.g., for observing one or more grooves of the sample slide) are selectable for the multiple grooves of the sample slide. The device and / or sample may include a predetermined position or positions. The sample holder may be configured to slide laterally to select a position for observing one or more grooves of the sample slide. This allows simple and easy selection of one of the grooves (e.g., four) of the sample slide (e.g., for observation).

[0054] The device may include at least one sensor. The at least one sensor may report measurements of at least one of the following: the position of the heating plate, the temperature (corresponding to the temperature of the sample), and the position of at least one objective lens. The sensor may report the measurements to a processing unit (e.g., software installed on a laptop). The temperature may be displayed on a user interface. The temperature may be updated in real time every few seconds.

[0055] The device may comprise a mechanism for moving the sample holder relative to at least one objective lens. This may be for focusing on the sample. The mechanism may comprise a focus knob configured to rotate to move the sample holder. The device may comprise a means for alerting a user when the sample holder reaches a limit of the mechanism (e.g., the base of the mechanism). The means for alerting the user may be a microswitch that is activated when the sample stage reaches a limit (e.g., a lower limit) and a pop-up window may be displayed on the screen (or more generally on the user interface) to inform the user that the user needs to move the sample stage in the other direction. The microswitch may send a value of 0 if it is safe, or a value of 1 if the limit is being approached. The value of the microswitch may be updated in real time every few seconds, which is important when focusing on the sample. This has the advantage that the user is notified when the limit is reached and does not attempt to continue lowering the sample stage beyond the limit of the mechanism (e.g., if they are unaware that they are moving it in the wrong direction). If the user continues to lower the sample stage beyond the limit of the mechanism, the gear system may break and the device may become inoperable.

[0056] The processing unit or a processing unit in the device may be configured to at least control the current to the LEDs, control the current to the heating stage, collect signals related to the time and / or position of the measurement (e.g., by collecting GPS signals) and / or warnings that the sample stage has reached a limit of the mechanism (i.e., position sensors), and / or signals related to position sensors that measure the position of the first objective lens, the second objective lens and / or the objective lens mounting plate. The processing unit may be located on the PCB.

[0057] There may be a wireless connection between the processing unit and the cloud where the time-correlated intensity variations of the image at each q are analyzed to generate parameters that characterize the particles.

[0058] The sample holder, the objective lens translation stage, and / or the mechanism for moving the sample holder relative to at least one objective lens (e.g., focus knob) may be accessible from outside the box. The sample holder can be slid laterally to allow external access to the location of the sample slide (e.g., to add, remove, or replace a sample slide) or to remove the sample holder completely. The objective lens translation stage can be slid laterally to switch objective lenses. The focus knob can be rotated to move the sample holder.

[0059] The device may include a chassis configured to hold the components of the device together. The chassis and / or the components of the device may be 3D printed and / or machined. More generally, at least some of the components of the device may be 3D printed.

[0060] According to a third aspect of the invention, there is provided a method of measuring at least one parameter of a particle with a portable device, the method comprising providing a sample comprising particles in solution, introducing the sample into a sample holder in the portable device, and generating an image (e.g. a digital image) from an imaging means (e.g. an imaging sensor) in the portable device (e.g. in a camera). In an embodiment, the method further comprises recording the digital image at a frame rate of greater than 100 frames / s, preferably greater than 200 frames / s, more preferably greater than 300 frames / s.

[0061] The method can include processing the digital images in a processing unit in the portable device or in a remote processing unit by performing differential dynamic microscopy. This can include analyzing the power spectrum of the difference between pairs of spatial Fourier transforms of the digital images separated by a time delay over a range of time delays and spatial Fourier frequencies. This can be over all time delays and spatial frequencies or any subsection. This can be over all possible Fourier image pairs or selected Fourier image pairs and / or all possible q-values ​​and / or selected q-values.

[0062] The method may include selecting a pixel size for the image from the range of 0.1 microns / pixel to 10.0 microns / pixel, preferably 0.5 microns / pixel to 7 microns / pixel.

[0063] According to a fourth aspect of the invention, there is provided a portable device or system according to any other aspect and / or embodiment described herein, wherein the device is configured such that light from the light source is transmitted to the sample holder without undergoing refraction.

[0064] According to a fifth aspect of the invention, there is provided a portable device or system according to any other aspect and / or embodiment described herein, wherein the device comprises an objective lens movement stage that slides laterally to switch between at least two objective lenses.

[0065] According to a sixth aspect of the invention there is provided a portable device or system according to any other aspect and / or embodiment described herein, wherein the device comprises a heating stage and / or cooling stage configured to heat, cool and / or maintain the temperature of particles in solution, the sample holder and the heating stage and / or cooling stage being configured such that a slide for holding the particles in solution is in contact with the heating stage and / or cooling stage in use. The heating stage and the cooling stage may be integrated into a (single) heating / cooling stage.

[0066] According to a seventh aspect of the invention there is provided a portable device or system according to any other aspect and / or embodiment described herein, wherein the device comprises at least one sensor for measuring at least one of a position of a heating plate, a temperature, and / or a position of at least one objective lens.

[0067] According to an eighth aspect of the invention there is provided a portable device or system according to any other aspect and / or embodiment described herein, wherein the imaging sensor has a pixel size in the range of 0.5 to 10 microns / pixel, more preferably in the range of 2 to 5 microns / pixel, and / or the imaging sensor has a frame rate greater than 50 frames / sec, preferably greater than 100 frames / sec, more preferably greater than 300 frames / sec.

[0068] According to a ninth aspect of the invention, there is provided a method for manufacturing a portable device for measuring at least one parameter of particles in a solution, the method comprising providing a light source, providing an imaging means (e.g. an imaging sensor) for generating an image (e.g. a digital image), providing an optical system comprising at least one objective lens, and providing a sample holder.

[0069] The method may include arranging the optical system and the imaging sensor such that a pixel size of the image is in at least one of the ranges of 0.1 microns / pixel to 10.0 microns / pixel, preferably 0.5 microns / pixel to 7 microns / pixel.

[0070] According to a tenth aspect of the invention there is provided a computer program comprising computer readable instructions configured to cause a computer to carry out the method of any aspect or embodiment.

[0071] According to an eleventh aspect of the invention there is provided a computer readable medium carrying a computer program according to the tenth aspect.

[0072] According to a twelfth aspect of the invention there is provided a computing apparatus comprising a memory storing processor-readable instructions and a processor configured to read and execute instructions stored in the memory, the processor-readable instructions including instructions configured to control a computer to perform a method of any aspect or embodiment.

[0073] Advantages include an easy-to-use, automated, portable instrument for semen analysis, it can provide reproducible motility measurements (error <5%), and the method can be valid for any semen concentration above 1 million / mL (below sex-sorted semen concentration).

[0074] Features in one aspect may be applied as features in any other aspect in any suitable combination, for example, features of an apparatus, device, and / or system may be applied as features of a method, and vice versa, in any combination.

[0075] It is to be understood that each individual feature and / or combination of features defined above according to any aspect of the invention or below in relation to any particular embodiment of the invention can be used separately and individually, alone or in combination with any other defined feature, in any other aspect or embodiment of the invention.

[0076] Furthermore, the present invention is intended to cover apparatus, devices and / or systems configured to carry out any of the features described herein in relation to the methods, and / or methods of using or producing, using or manufacturing features of any of the apparatus, devices and / or systems described herein. [Brief description of the drawings]

[0077] Embodiments of the present invention will now be described, by way of example only, with reference to the accompanying schematic drawings, in which: [Figure 1] FIG. 1 shows a perspective view of a device for measuring parameters of particles in a solution according to one embodiment of the present invention. [Diagram 2] 1 shows a box for a device according to one embodiment of the present invention. [Diagram 3] 1 shows a perspective longitudinal cross-sectional view of a device according to one embodiment of the present invention. [Figure 4] FIG. 2 shows a cross-sectional view of a device according to one embodiment of the present invention. [Diagram 5] 2 shows a cross-sectional view of the mirrors, lenses, and camera of a device according to one embodiment of the present invention. [Figure 6] 1 shows a differential image correlation function for a lag time τ=0.01 seconds, according to one embodiment of the present invention. [Figure 7] 1 shows a differential image correlation function for a lag time τ=0.01 seconds, according to one embodiment of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0078] FIG. 1 shows a device 10 for measuring parameters of particles in a solution (i.e. a sample). The device 10 may be a microscope or an imaging module. The device 10 comprises a light source that illuminates the sample, which in this embodiment is an LED 12 in an LED holder 14. In an embodiment, the LED 12 may be green. This may be for bright field and / or phase contrast imaging of the sample. It will be understood that in other embodiments the LED may be any color and the light source may be different from the LED.

[0079] The particles in the solution can be microorganisms. The particles in the solution can be colloids, droplets (e.g., milk fat droplets), and / or emulsions. The microorganisms can be sperm, bacteria such as Escherichia coli, also known as E. coli, and / or microalgae. The microorganisms can vary in size from bacteria to microalgae.

[0080] The device 10 may be for measuring a single parameter of a particle or multiple parameters of a particle. The parameters of a particle may include the particle's motility, preferably the rate of movement, the average velocity, the concentration, the size, the amplitude of the head movement, the diffusivity, and / or the frequency of the head movement. More generally, the intensity variations in the image are characterized. The intensity variations in the image are due to the particle movement and therefore contain all the information of the particle movement. More specifically, the spatiotemporal variations across all the images on the video are characterized. In other words, "characterization of the spatiotemporal variations of the intensity across the images" or "the variations of the intensity in space and time across the images".

[0081] The device 10 extends horizontally in an xz plane, perpendicular to the y direction, as shown. For ease of understanding, the z direction will be referred to as horizontal and the x direction as vertical. It will be understood that this is merely by convention and is not limiting. For example, the device may be generally vertical, horizontal, or two horizontal. For example, rather than the y axis as shown in the figure, the z axis may instead extend vertically.

[0082] The device 10 includes a sample stage 16, which includes a sample holder 18 that is slidably movable relative to the sample stage 16. That is, the sample holder 18 can slide laterally within the sample stage 16. The sample holder 18 is configured to hold a sample slide 20. The sample holder 18 can be configured to slide laterally to select a position for observing one or more grooves 21 of the sample slide 20. For example, the sample slide 20 can include four grooves 21, and there can be positions for observing the grooves 21 that can be selected relative to the sample slide 20. This does not require the user to directly look at the grooves. There can be a means of feedback to the user that the sample slide is correctly in the observation position (e.g., there can be an audible or tactile click). When the groove is in the correct position under the LED 12, the groove is considered to be in the observation position within the observation area. The correct position can be directly under the LED, or it can be an intermediate component (e.g., a light diffuser) between the LED and the sample, which in this case does not need to be directly considered. It will be appreciated that in other embodiments, there may be more or less than four grooves on the specimen slide.

[0083] The device 10 is configured such that the light from the LED 12 is transmitted to the sample holder 18 without refraction. That is, the device 10 does not include a focusing lens. The device 10 is configured such that the light from the LED 12 is incident on the sample in the sample holder 18 without refraction. In other words, the light is incident directly from the LED 12 on the sample in the sample holder 18. This has the advantage that the device 10 can be physically smaller and lighter (i.e., easier to carry or move). The sample holder 18 can hold the size of a typical microscope sample slide (75×26 mm and 1 mm thick (imperial) or 75×25 mm (metric) and 1 mm thick), although the samples can be smaller or larger, thinner or thicker. In other embodiments, the sample holder can be sized to hold different sized samples and / or different sized sample slides.

[0084] Standard microscopes include a focusing lens to focus light onto the objective lens. This is because the microscope is intended to be used at high magnifications that require more light. A focusing lens is not required for device 10 because device 10 is generally based on low magnifications and therefore does not require high illumination. Avoiding the use of a focusing lens in device 10 may not adversely affect imaging and analysis, but may allow for a reduction in the overall height of device 10 of approximately 5 cm, which may represent a 25%-30% reduction in height (i.e., making it physically smaller).

[0085] The device 10 comprises an optical system including two objective lenses, namely a first objective lens 22 and a second objective lens 24. The first objective lens 22 has a lower magnification than the second objective lens 24. The objective lenses 22, 24 are fixedly arranged in an objective lens mounting plate 26 (or an objective lens moving stage), which is arranged in an objective lens mounting clamp 28. The objective lens mounting plate 26 is slidably movable relative to the objective lens mounting clamp 28. The objective lens mounting plate 26 can be called a slider. That is, the objective lens mounting plate 26 can slide laterally in the objective lens mounting clamp 28 to switch between the first objective lens 22 and the second objective lens 24. The objective lens mounting plate 26 can be arranged so that light reflected from the sample passes through either the first objective lens 22 or the second objective lens 24 (depending on which objective lens is in the correct position). The objective lenses 22, 24 need to be precisely positioned (i.e. aligned with the rest of the optical setup) for effective imaging. This means that the objective lenses 22, 24 must be returned to the correct position. The device 10 may include means for returning the objective lenses 22, 24 to the correct position. The means for returning the objective lenses 22, 24 to the correct position may include a spring ball (not shown) and / or a recess (not shown) in the objective lens mounting plate 26.

[0086] In embodiments, the objective lens mounting plate 26 may be moved manually (e.g., by a user manually) and / or controlled (e.g., electronically, e.g., by a processing unit) and / or automated. Additionally, the means for returning the objective lenses 22, 24 to the correct position may be manual and / or controlled and / or automated.

[0087] Automatic detection of the position of the objective lens mounting plate 26, i.e., which objective lens 22, 24 is selected for imaging, can be provided so that the software can automatically detect the selected imaging mode. This can be done by position sensors (not shown) that measure and report the position of the first objective lens 22, the second objective lens 24 and / or the objective lens mounting plate 26.

[0088] The device 10 comprises a mechanism for moving the sample holder 18 relative to the objective lenses 22, 24 (in practice, the sample stage 16 is moved and the sample holder 18 moves with it). This may be to focus the sample. The mechanism comprises a focus knob 30 configured to rotate to move the sample holder 18. The knob 30 is attached to a rod 32 which, via a gear system (not shown), rotates a lead screw 34 which moves a lead screw nut (not shown) up and down. The lead screw nut is attached to the sample stage 16, meaning that the sample stage 16 moves up and down as well. The total travel distance the sample stage 16 can travel may set the depth of focus range for DDM processing and visual imaging.

[0089] The device 10 may be provided with means to warn the user when the sample holder 18 (actually the sample stage 16) reaches a limit of the mechanism (e.g. the base of the mechanism). The means to warn the user may be a microswitch (or more generally a position sensor) that is activated when the sample stage 16 reaches a limit (e.g. the lower limit) and a pop-up window may be displayed (for example) on the screen to inform the user that the user needs to move the sample stage 16 in the other direction. This has the advantage that the user is informed when the limit is reached and that the user does not try to continue lowering the sample stage beyond the limit of the mechanism (e.g. if he does not realize that he is moving it in the wrong direction). If the user continues to lower the sample stage beyond the limit of the mechanism, the gear system may break and the device 10 may become inoperable. This may be used to avoid contact between the heated stage 52 (see FIG. 3) and one of the objectives 22, 24 (e.g. an objective not being used during imaging).

[0090] The sensors can report measurements to a processing unit, which can be provided to a user (eg, using a screen).

[0091] The base housing 36 is disposed at the bottom of the device 10 and surrounds other components of the device 10, which will be described later. The base housing 36 may be for ensuring a relatively closed environment, for example, to prevent dust from accumulating in the optical system or to prevent external light from affecting imaging. The two guide rods 38 may extend vertically substantially the entire height of the device 10 and hold the components of the device 10 together. However, in other embodiments, other methods may be used to enable y-axis movement. The base housing 36 and / or the two guide rods 38 may form part of the chassis of the device 10. The chassis may include other parts of the device 10, not specifically described, that may hold the components of the device together. The chassis and / or other components of the device 10 may be 3D printed and / or machined. For example, all components of the microscope, except for the rotating elements of the y-axis subsystem, the y-axis guide rods, and the heating stage, may be 3D printed. A significant portion or at least half of the mass of the device 10 may be in the lower quarter of the device 10. This has the advantage of helping to prevent the device from tipping over.

[0092] FIG. 2 shows the device 10 enclosed within a box 40. The box 40 may be considered to form part of the device 10, and the box 40 may be considered to substantially completely enclose the components of the device 10 (except as described below). As can be seen, a portion of the sample holder 18, a portion of the objective lens mounting plate 26, and the focus knob 30 are visible and accessible from outside the box 40. The sample holder 18 and the objective lens mounting plate 26 pass through slots in the box 40. The knob 30 can be rotated from outside the box 40 to move the sample holder 18 up and down. As can be seen, the slots in the box 40 around the sample holder 18 extend in the vertical (y) direction to allow the sample holder 18 to move up and down. Furthermore, the sample holder 18 may be slid laterally away from the box 40 to allow external access to the location of the sample slide 20 (e.g., to add, remove or replace the sample slide 20) or to remove the sample holder 18 completely. Additionally, the objective lens mounting plate 26 can be slid laterally away from the box 40 to switch from the first objective lens 22 to the second objective lens 24. Similarly, the objective lens mounting plate 26 can be slid laterally towards the box 40 to switch from the second objective lens 24 to the first objective lens 22. Thus, switching between the first objective lens 22 and the second objective lens 24 can be performed outside the box 40.

[0093] In an embodiment, some of the components of the device 10 may be motorized and / or an automation system may be present. For example, these components may include the sample stage 16, the sample holder 18, and / or the objective mounting plate 26. This may mean that parts of the sample holder 18 and / or parts of the objective mounting plate 26 and / or the focus knob 30 that are outside the box 40 that can be held by the user may not be needed. In this case, the user may not need to select the sample holder 18, the objective mounting plate 26, and / or the focus knob 30 outside the box 40 (other than inserting the sample slide 20 into the box 40). This may mean that the box 40 is more enclosed and protected from the environment.

[0094] Box 40 may be made from a material that is waterproof and / or dustproof and / or relatively durable (e.g., aluminum, various commodity plastics (PP, glass filled, HDPE, ABS, PC / ABS blends, TPE overmolding)). Advantages of enclosing the components of device 10 within box 40 include: the components can be kept dry (preventing condensation on optical components), kept clean, protected from light and dust, etc., protected from physical damage, and not contaminated by the environment (e.g., location in a farm). Additionally, a user cannot access the components to change settings, etc., which may result in inaccurate results. Box 40 also adds a user interface point for moving, storing, operating, transporting device 10. Box 40 may also provide EMC shielding. Additionally, more components (such as electronic components) may be added to the box, which helps provide a safe environment for the user and / or improve EMC shielding. Device 10 and / or box 40 may include vibration or shock absorbing materials. Vibrations can be detrimental to microscopic imaging and having vibration or shock absorbers can help mitigate this.

[0095] Box 40 includes an on / off switch 42. In this embodiment, device 10 is powered from a power source and box 40 includes a connection 44 for power from the power source. In other embodiments, device 10 may be battery powered (battery not shown). In some embodiments, device 10 may have both a power source and battery power option.

[0096] Box 40 also includes a USB connection 46 for transferring data from device 10 (i.e., outside of box 40). This can include data from an imaging sensor and / or camera. This can also include data from any other components that can provide, for example, a GPS signal, and / or from an electronics board that collects temperature readings from sample stage 16 into software for real-time reading of sample temperature. It will be appreciated that this is only one example and other means for transferring data from device 10 (outside of the box) can be used. In other embodiments, there is no USB connection, data can be transferred wirelessly, or processing of data can occur within box 40. In an embodiment, device 10 can be operated using a Windows 10 laptop.

[0097] Box 40 also includes a GPS locator cap 48, under which is a GPS antenna (not shown). Cap 48 can be 3D printed. The GPS antenna does not have to be enclosed within box 40 (which can be metallic) as otherwise there may be no signal (or at least a reduced signal). A PCBA is also provided to collect the signal from the GPS antenna. It will be appreciated that in other embodiments there may be other ways of detecting the location of box 40 (and where measurements are taken).

[0098] The device 10 is portable. For example, the device 10 can be moved by hand by a person and is relatively small and lightweight. The device 10 (including the box 40) can have dimensions of about 30×20×15 cm, and more typically can be smaller than 40×30×20 cm. The device 10 (including the box 40) can be less than 5 kg, and more preferably the box can be 2.5 kg. For example, the box 40 can be about 4 kg. The box 40 can include a handle 50 for easy carrying. The device 10 does not need to be used in a laboratory environment (even though it can) and does not need to be disassembled into components to be moved.

[0099] Box 40 also includes four legs 49 (only three shown). These legs 49 may be anti-vibration legs that may help provide a good video image. It will be appreciated that in other embodiments there may be more or less than four legs. More generally, in embodiments the box may include multiple legs, and more preferably multiple anti-vibration legs.

[0100] 3 is a perspective cross-sectional view taken vertically (i.e., along the x-direction) through device 10. Components of device 10 that were hidden behind base housing 36 in FIG.

[0101] A cross section of the sample stage 16 is shown. The device 10 also includes a heating stage 52. The heating stage 52 is configured to heat and / or maintain the temperature of the sample at a predetermined temperature by heating and / or maintaining the temperature of the sample slide 20 holding the sample. The predetermined temperature can be between ambient temperature (e.g., 15 to 25° C.) and 50° C. There can be a maximum temperature that is set depending on the material surrounding the heating stage 52 (e.g., based on the maximum temperature of the 3D printed material). The sample and / or the sample slide 20 can be heated and / or maintained at substantially 37.5° C. + / - 0.5° C. (e.g., for bacteria or sperm). The sample can be maintained at other temperatures depending on the microorganism or colloid. In an embodiment, alternatively or additionally, the device 10 can include a cooling system (e.g., to cool and / or maintain the temperature of the sample at a predetermined temperature). This can include a Peltier module. This allows for more precise control of the temperature over a wider temperature range.

[0102] The heating stage 52 comprises a plate 54 (e.g. made of aluminium) which is heated to a predetermined temperature. The heating stage 52 may comprise resistors (not shown) arranged in two side holes 56 for heating the plate 54. There may be two resistors. There may be six resistors in series, two in each side hole. This may provide a relatively good uniformity of temperature across the plate. There may also be a number of resistors different from two or six. The heating stage 52 may comprise a temperature sensor (not shown) for measuring the temperature of the plate 54. The temperature sensor may be arranged in a central hole 58 of the heating stage 52. The temperature may be read continuously during use and the temperature measurements may be available in real time. Measurements may be made, for example, every second.

[0103] The sample holder 18 and the heating stage 52 are configured such that the sample slide 20 comes into contact with the heating stage 52 (i.e., the plate 54). This can be by the plate 54 lifting the sample slide 20. This can be because the top surface of the plate 54 is above the surface of the sample holder 18 on which the sample slide 20 rests. In other words, the side of the (3D printed) sample holder 18 can be lower than the top surface of the plate 54 so that the sample slide 20 only contacts the heating plate 54 when it is slid in. There may not be a spring or active force pushing the sample slide 20 into the heating plate 54. They may simply be in nominal contact and rely on gravity to maintain contact. There may be a vertical constraint on the sample slide 20 using a lock on the sample holder 18 and a lip on the sample holder 18. However, these features are more to aid in manual manipulation of the sample before it enters the device 10 than to maintain contact with the heating plate.

[0104] Device 10 also includes a mirror 60 held in a mirror mount 62. Mirror 60 may be considered to form part of the optical system of device 10.

[0105] FIG. 4 shows a cross-section taken laterally (i.e. along the z-direction) through the device 10, showing the LED 12, the first objective lens 22 and the mirror 60 aligned with each other. This means that the light from the LED 12 is incident on the sample in the sample slide 20 (the sample is in the groove 21 of the sample slide 20 located in the observation area). The light then passes through the sample and enters the first objective lens 22, which sends the light to the mirror 60, from which it is then reflected. The mirror 60 reflects the light at about 90 degrees (i.e. from vertical to horizontal, out of the page as shown). The use of the mirror 60 means that the height of the device 10 can be reduced. The mirror 60 can be provided to fit the components of the device 10 into a stable and conveniently sized box 40. It will be understood that in other embodiments the mirror 60 is not required.

[0106] Referring again to FIG. 3, the light reflected from the mirror 60 is incident on a lens 64 (not shown in FIG. 3, but see FIG. 5) in a lens holder 66. The lens 64 is considered to form part of the optical system of the device 10 and can be called a further lens. The lens 64 focuses the light on an imaging sensor 68 (see FIG. 5) in a camera 70. The lens can allow for parfocal imaging, i.e. allowing (approximately) the same focal plane when changing the objective lens and / or fine-tuning the size of the pixels of the resulting image by moving the lens in the horizontal plane, and therefore the lens can be called an adjustment lens. The imaging sensor 68 is for generating a digital image. The camera 40 can be a digital camera (including the imaging sensor 68) embedded in a camera mount that can be made of 3D printed material. The material surrounding the camera mount can allow for ventilation to avoid overheating of the camera 70. The position of the camera 40 (and thus the imaging sensor 68) may be adjustable or adjusted in the x-plane to fine-tune the size of the pixels in the resulting digital image.

[0107] Figure 5 shows a cross-sectional view taken vertically (i.e., along the x-direction) through a portion of device 10. Mirror mount 62, mirror 60, lens 64, lens holder 66, camera 70 and base housing 36 are all shown in Figure 5. Lens 64 may be a plano-convex lens.

[0108] The sensors can report measurements to a processing unit, which can be provided to a user (eg, using a screen).

[0109] The device 10 may include a processing unit (not shown) in the box 40 for processing the digital images to obtain parameters of the particles in the solution. In other embodiments, there may be a system including the device 10 and a remote processing unit for processing the digital images to obtain parameters of the particles in the solution. In embodiments, the processing unit and / or the remote processing unit may include 1) a laptop computer (e.g., using Windows 10), 2) a desktop computer, 3) embedded processing in the unit, 4) a tablet or smartphone, 5) the cloud, 6) a combination of one or more of 1) to 5). There may be a wired or wireless connection between the device 10 and the processing unit and / or the remote processing unit.

[0110] Parameters of the particles in solution (i.e., the sample) can be obtained by a processing unit (within box 40 or remote from box 40) analyzing (or calculating) the power spectrum of the difference between pairs of spatial Fourier transforms of digital images separated by a time delay over a range of time delays and spatial Fourier frequencies. This can be done by digitally processing the images by the Fourier transform of each image such that the wave vector q represents the spatial-domain and time-correlated intensity variations of the image at each q. This can be referred to as differential dynamic microscopy (DDM).

[0111] Analyzing the power spectrum of the difference between pairs of spatial Fourier transforms of digital images separated by a time delay over a range of time delays and spatial Fourier frequencies can include: (1) calculating the differential image correlation function (DICF), i.e., calculating the spatial Fourier transform of the image, and then calculating the power spectrum of the difference between pairs of Fourier images over the range of available delay times τ (i.e., the time difference between two selected images) and spatial frequencies (q) provided by the Fourier transform of the image. All possible Fourier image pairs or selected Fourier image pairs can be considered. The calculation can be done for all possible Fourier image pairs or a subsection of the Fourier image pairs. (2) Then averaging all the resulting DICFs that have the same delay time (τ) together, and (3) then performing radial averaging for each q value to generate the final time-averaged and vector {q}-averaged DICFs as a function of delay time τ and spatial frequency q. All possible q values ​​or selected q values ​​can be considered. The calculation can be done for all possible q values ​​or a subsection of q values. DDM includes calculating the power spectrum of the difference between pairs of images. This process involves the calculation of a Fourier transform that defines the Fourier component q, which defines the length scale. The spacing between pairs of images defines the delay time τ. In practice, the range of q values ​​is defined by the image size in pixels and the size of the pixels in the image. The standard method is to calculate the Fourier transform for all possible q values, which may correspond to half the number of pixels in the image. The total number of Fourier image pairs can be calculated by N*(N-1) / 2, where N is the number of images. Therefore, considering all possible pairs and all possible q values ​​requires significant computer processing power and memory to store all the calculations in memory. Taking a sub-selection (subsection) of the number of Fourier image pairs (e.g. considering only one tenth of the image pairs) and q values ​​(e.g. about 20 values ​​compared to the 256 q values ​​expected for an image of 512x512 pixel size) means that processing times can be reduced.For example, processing time can be reduced from about 10 minutes (e.g., using a laptop with 16 Gb RAM) to about 2 minutes, and memory usage can be reduced from about 12 GB to about 5 Gb for a video of 10,000 images with an image size of 512x512 pixels. Importantly, this allows the use of lower spec processing units, e.g., laptops with 8 Gb RAM rather than 16 Gb laptops. In embodiments, the subsection (or subselection) of Fourier image pairs and / or q values ​​can be calculated using an algorithm, or can be user selected, or can be otherwise determined. The minimum number of possible Fourier image pairs per delay time used for the size of the subsection can be 10. This can, for example, give results within 10% of the true result. The minimum number of Fourier image pairs per delay time can depend on the concentration of the particles and the size of the particles.

[0112] The digital image processing and imaging technique can be carried out by differential dynamic microscopy (DDM). The digital image processing and imaging technique can be carried out using any function or any method described in (High-throughput characterisation of bull semen motility using differential dynamic microscopy (plos.org) (PLoS ONE 14(4): e0202720. https: / / doi.org / 10.1371 / journal.pone.0202720) and Differential Dynamic Microscopy: A High-Throughput Method for Characterizing the Motility of Microorganisms (Biophysical Journal Volume 103 October 2012 1637-1647), both of which are incorporated herein by reference.

[0113] There can be a wireless connection between the processing unit (wherever it is located) and the cloud where the time-correlated Fourier image intensity fluctuations at each q are analyzed to generate parameters that characterize the particle.

[0114] The processing unit may be configured to control the current to the LED 12, control the current to the heating stage 52, and collect (e.g., by collecting GPS signals) signals related to the time and / or position of the measurement, and / or a warning that the sample stage 16 has reached a limit of its mechanism (i.e., position sensors), and / or signals related to position sensors measuring the position of the first objective lens 22, the second objective lens 24, and / or the objective lens mounting plate 26. The GPS signals may provide an accurate time stamp so that continuous measurements of motility parameters can be made over time. The location of the measurement (of a particle parameter) may also be determined using the GPS signals to identify where the sample (e.g., semen) was tested and to investigate correlations between the measurements and other factors, e.g., weather, diet, environmental conditions, etc. The processing unit may be located on the PCB.

[0115] The device 10 or system may comprise means for displaying a parameter of the particles in the solution. The means for displaying a parameter of the particles in the solution may be a screen (not shown).

[0116] As mentioned above, the two objective lenses 22, 24 each have a different magnification. The first objective lens 22 is for enabling an image to be captured for DDM processing (e.g., for measuring the motility of the microorganisms). The second objective lens 24 is for visual inspection of the particles (e.g., for visualizing the particles in the solution with a resolution in which the size of the image of the head of the microorganism is in the range of 5 to 15 pixels). It will be understood that in other embodiments, the first objective lens can be for visual inspection and the second objective lens can be for DDM processing. In other embodiments, there can be a single objective lens that can be for example only for DDM processing. In embodiments, both the first objective lens and / or the second objective lens can be used for further image processing other than DDM.

[0117] In an embodiment, both the first objective lens 22 and the second objective lens 24 may be used for DDM processing, albeit for different sized particles in the solution. For example, the first objective lens 22 may be used for DDM of sperm, and the second objective lens 24 may be used for visual inspection of sperm and DDM of different sized particles (such as bacteria and algae). Thus, an objective lens may be used for more than one purpose.

[0118] The first objective lens 22 and / or the second objective lens 24 (e.g., specifically for sperm) may have a sufficiently large depth of field such that the resulting digital image includes all cells traversing a vertical cross section of the groove, e.g., a 20 micron high groove, which may eliminate the need to collect multiple images at different focal planes through the sample.

[0119] The first objective lens 22 and / or the second objective lens 24 may provide a relatively large field of view so that a large amount of the sample may be imaged, which may eliminate the need to collect multiple images across different locations of the sample for better statistics.

[0120] The optical system and the imaging sensor 68 are configured such that the image pixel size can be selected for different particle sizes. That is, the magnification of the first objective lens 22 and / or the second objective lens 24, the distance between the objective lenses 22, 24 and the imaging sensor 68, and the size of the pixels of the imaging sensor 68 are each selected such that the image pixel size (i.e., the resolution) is predetermined or falls within a predetermined range. Thus, the device 10 can be preconfigured to provide a desired image pixel size.

[0121] As an example, the first objective lens 22 may have a magnification such that the size of the head of a microorganism (eg, sperm) in an image is in the range of 1 to 5 pixels.

[0122] As other examples, the first objective lens 22 and / or the second objective lens 24 may have a magnification in the range of 1x to 4x (e.g., for sperm), in the range of 5x to 10x (e.g., for bacteria such as E. coli), and / or in the range of 5x to 20x (e.g., for microalgae).

[0123] The pixel size of the image can be substantially 0.9 microns / pixel, 1.7 microns / pixel, 2.1 microns / pixel, and / or 4.3 microns / pixel. For example, 0.9 microns / pixel, more specifically 0.86 microns / pixel, allows imaging of sperm and / or microalgae flagella (e.g., approximately 1 micron thick). 4.3 microns / pixel allows performing DDM on sperm and / or microalgae. 0.9 microns / pixel, 1.7 microns / pixel, and / or 2.1 microns / pixel allows performing DDM on bacteria.

[0124] The imaging sensor 68 may have a pixel size in the range of 0.5 to 10 microns / pixel, and more preferably in the range of 2 to 5 microns / pixel.

[0125] The imaging sensor 68 can be configured to operate in 1x1, 2x2 and 4x4 binning modes. The imaging sensor 68 can be configured to operate in a skipping mode.

[0126] Once the components of device 10 are enclosed within box 40, the user can only switch the objective lenses 22, 24 to switch between different modes (e.g., viewing and DDM processing) and cannot otherwise change the settings to change the pixel size of the image.

[0127] The imaging sensor 68 may have a frame rate of greater than 50 frames / sec, preferably greater than 100 frames / sec, and more preferably greater than 300 frames / sec. A higher frame rate may enable characterization of faster particles, e.g., small particles having a size less than 100 nanometers in diameter, or fast swimmers, e.g., greater than 50 microns.

[0128] In use, a user adds a sample containing particles in solution to groove 21 of sample slide 20, then places sample slide 20 into sample holder 18 (which has been slid away from box 40 for access), and then slides sample holder 18 into box 40 until the user hears a click that tells the user the groove is in the correct position to be observed.

[0129] The first objective lens 22 and / or the second objective lens 24 are selected based on the desired imaging, i.e., the pixel size (microns / pixel) of the image is set based on the purpose. For example, if it is desired to perform DDM on sperm, the objective lens mounting plate 26 can be slid laterally toward the box 40 so that the first objective lens 22 is in the observation position.

[0130] Light from LED 12 illuminates the sample, and the light from the sample then passes through first objective lens 22, reflects off mirror 60, is refracted (e.g., focused) by lens 64, and then impinges on imaging sensor 68. The sample stage 16 is then moved up and down to focus on the sample. The user can view a screen with a live stream of the imaging, and then adjust focus knob 30 accordingly.

[0131] The imaging sensor 68 produces a digital image, which is then processed by a processing unit that analyzes (or calculates) the power spectrum of the difference between pairs of spatial Fourier transforms of the digital images separated by a time delay over a range of time delays and spatial Fourier frequencies to obtain parameters of the particles in solution (e.g., sperm motility). The results can be presented to the user on a screen.

[0132] The data collection and / or data analysis can be automated to generate measurements of parameters of the particles. In embodiments, the device and / or system can include user interaction means (such as a button) for performing the data collection and / or data analysis. The data collection and / or data analysis can be performed automatically by a user clicking a button (e.g., once). A measurement or a series of measurements can be performed over time (e.g., up to 12 hours) (in real time) by a user simply clicking a button (i.e., a single user interaction).

[0133] In an embodiment, the image pixel size may be in the range of about 2 to 7 microns / pixel. This is believed to be the optimal range, as explained below. For clarity, all pixels in an image may have the same size, but the size of the image pixels may be adjusted by the optical system, imaging sensor, etc. For DDM to be useful, for example, for sperm motility, it may be necessary to achieve low magnification to reach the required low q value. A low q value is <0.4 μm -1 , or 0.05 μm -1to 0.4 μm -1 Low magnification would be considered to mean that the image pixel size is between about 2 microns / pixel and 7 microns / pixel. To illustrate why the range of 2 microns / pixel to 7 microns / pixel is considered to be the optimal range (e.g., for sperm motility), three example image pixel sizes (a), (b) and (c) and the results for these sizes are shown below. Example (a) is at the low end of the 2 to 7 microns / pixel range, example (c) is at the high end of the 2 to 7 microns / pixel range, and example (b) is about halfway between the 2 to 7 microns / pixel range.

[0134] Importantly, for example, the DDM applied to the measurement of sperm motility has a resolution of about 0.05 μm. -1 to 0.4 μm -1A range of q values ​​may be needed, with the final range being determined by the animal species. The available q values ​​depend on both the pixel size of the image (i.e. image pixel size) and the size of the image in pixels (image size in pixels). The Q value is the true value of the spatial Fourier frequency q and has the dimension of the inverse of the length (e.g. here 1 / micron). The Q index is the number of pixels along a radial line of the Fourier image where the radial averaging is performed. An example is given with the aid of Figure 6 to explain what the q index means. When calculating the differential image correlation function (DICF), there is one image per DICF for a given delay time. Figure 6 shows the DICF for a delay time τ=0.01 seconds. This resulting image is 251 pixels wide and 500 pixels high. Then the radial averaging is performed. This means that all the values ​​of this image are averaged along a semicircle, shown as a black line. The radius of this line can be defined as the number of pixels along a radial line from the center of the image. Thus, q_index=0 means the first pixel in the center of the image. Q_index=250 is the last pixel on the right side. The true value of q depends on the radius used to compute the radial average, so it requires knowing the pixel size over which the radial average is done (defined by q_index) and the total size of the image in pixels. The formula to compute the q value from the q index is:

[0135] Equation 1: Q = 2 * π * q index / (image pixel size * image size in pixels). For example, pixel size = 4.3 μm, image size in pixels = 512 pixels, q index = 80 → q = 0.228 μm -1 It is.

[0136] A low q-index may result in a noisier differential image correlation function (DICF) since there are fewer pixels to perform radial averaging to obtain the final DICF, making it more difficult to measure sperm motility. A low q-index has a smaller radius, so the radial average is performed over a smaller number of pixels. An example is given with FIG. 7 to explain what a low q-index means. A low q-index means that there are fewer pixels, since the radius of the semicircle is smaller and therefore its circumference is smaller. A simple example: Considering the first q_index (called q_index=0 in FIG. 7), at such a q_index there is only one pixel (located at the center left of the image). Considering the last q_index (called q_index=250 in FIG. 7), the radius of the largest semicircle is the largest possible in this image and has many pixels. In an embodiment, the lower limit of q_index may be 20 (which may be a good practice) or 10 may be used.

[0137] In the following example, images are limited to 512x512 pixels or less. This has to do with reducing the computer RAM and processing time required to process the images; i.e., processing larger images requires more computer RAM and longer processing time. For example, processing 10,000 images of 512x512 using a standard DDM algorithm may require a minimum of 12Gb of free RAM, with the processing time depending on the exact algorithm used. Using 1024x1024 pixels instead of 512x512 pixels in the image may require 12Gbx4=48Gb of free RAM. Using 512x512 pixels means that the total q index available is from 0 to 255. In practice, the increment of q_index is considered to be one pixel in the Fourier image. However, any increment at the sub-pixel level is considered in the Fourier image. Thus, for the example here, the increment is one pixel in the Fourier image. In an embodiment, all possible q values ​​are considered (and processed) for processing. Every possible q value corresponds to half the number of pixels in the image. However, in other embodiments, a subsection of all possible q values ​​is considered. For example, for a 512x512 pixel video, instead of taking all q indices from 0 to 255, only a range of q indices at a predefined step from 0 to 255 may be kept for the Fourier image. That is, a subsection of the full range of q indices may be selected, and within that subsection, only q indices every x (predefined step) may be kept. Using a subsection (or subselection) of all q indices means that the amount of RAM used can be reduced (e.g., relatively very large compared to processing all q indices). This allows the use of low-spec (or lower spec) processing units (e.g., laptops) and faster processing so that results can be obtained in less time (e.g., half the time or less). It will be understood that the absolute length of time depends on the computational power.The particular range of the subsections of the q index depends on the image size, and may be different ranges (and different predefined steps) for a 328x328 image, for example. It will be appreciated that there are many different possible subsection ranges and steps that can be used that can provide satisfactory results (e.g., obtain a parameter (such as head movement amplitude) to a desired level of accuracy). The smallest number of possible q values ​​considered for the subsection size may be 1.

[0138] Example (a): Image pixel size = 2 microns / pixel. This can be done by using the first objective lens 22 to capture images for DDM processing (e.g., to measure sperm motility). That is, using a magnification that is considered low (relative to standard microscopy). In other words, this setup is for DDM imaging and can be called DDM imaging mode.

[0139] The total field of view for a 512x512 is approximately 1mmx1mm. More precisely, with field of view = (image pixel size * image size in pixels), (2*512)^2 = (1.024)^2 mm^2.

[0140] ii. Using Equation 1, q = 0.05 to 0.4 μm -1 The q-index required to obtain is q-index = 8 to 65. For example, if q-index 65 is used, then q = 2 * π * 65 / (2 * 512) = 0.399 μm -1 to 0.4 μm -1 Having to use q-indexes between 8 and 65 means that we need to access the lower end of the q-index range, which is not ideal as it can increase the noise in the DICF.

[0141] iii. Use of the setup for visual imaging (can be referred to as visual imaging mode): This can be done by using the second objective lens 24 to capture images for visual inspection (e.g., of sperm). This can be based on a 10x magnification that can give an image pixel size of 0.4 microns, suitable for viewing individual flagella of sperm.

[0142] Example (b): Image pixel size = 4.3 microns / pixel, using DDM imaging mode.

[0143] i. The total field of view is approximately 2.2mmx2.2mm. More precisely, it is (4.3*512)^2=(2.2016)^2mm^2. This means that (compared to example (a), the field of view is approximately 4.62 times larger, so [(2.2016 / 1.024)^2=4.62]), so there are 4.62 times more cells in the field of view, and therefore we can average over more cells. For a given concentration, there are 4.62 times more cells than in example (a).

[0144] ii. Using Equation 1, q = 0.05 to 0.4 μm -1 The q index required to obtain is q index = 18 to 140.

[0145] iii.Furthermore, the image size can be reduced from 512x512 to 238x238, which gives the same final field of view as in example (a) since the image pixel size is larger, i.e. (4.3*238)^2=(1.024)^2mm^2, which is the same as in example (a). This allows for about a quarter less required free RAM and much faster DDM processing. In such a case the q index considered is the same as in example (a) [see point ii].

[0146] iv.Therefore, there is an advantage to using 4.3 microns / pixel for the image pixel size in terms of q-index selection for optimized DDM processing and improved overall size of the image.

[0147] v. Visual imaging mode (based on 10x magnification) can give an image pixel size of 0.86 microns, which is not as good as example a (see point iii), but is sufficient to see individual flagella.

[0148] Example (c) Image pixel size = 7 microns / pixel, using DDM imaging mode.

[0149] i. The total field of view is approximately 3.6mmx3.6mm. More precisely, it is (7*512)^2=3.584^2mm^2. Thus, the total field of view is 2.65 [(3.584^2) / (2.2016^2)=2.65] times larger than in the optimal case of example (b) [see point i].

[0150] ii. Using Equation 1, q = 0.05 to μm -1 The q-index required to obtain is q-index=28 to 228, which is an estimate, but is greater than 20 and therefore within the ideal range of q-indexes.

[0151] iii. However, due to inherent optical aberrations, maintaining all cells within the focal plane over such a large field of view may be difficult and ultimately not necessary.

[0152] iv. To give a similar field of view as the optimized case in example (b), the image size can be reduced to 316x316, which will reduce the amount of free RAM required and also speed up the DDM process.

[0153] v. However, visual imaging mode (based on 10x magnification) can give an image pixel size of 1.4 microns, which is not suitable for observing individual flagella as in example (b), but is still applicable.

[0154] All three examples (a), (b) and (c) are valid for DDM processing using the DDM imaging mode and for observing sperm flagella using the visual imaging mode, but each example has its advantages and disadvantages. For the visual imaging mode, it is possible to use a higher magnification (e.g., 20x magnification instead of 10x magnification), but this creates other physical problems, such as a shorter working distance from the top of the objective lens to where you want to focus in the sample. There are also specially large working distances for 20x magnification, but they are generally more expensive.

[0155] Although an image pixel size of 4.3 microns / pixel is mentioned (and believed to be particularly advantageous), it will be understood that there may be other values ​​(e.g., about 4.3 microns / pixel) that may be used and have the same or similar advantages (even if to a lesser extent). For example, in some embodiments, the image pixel size may be in the range of 4 to 4.5 microns / pixel.

[0156] As mentioned above, there are many factors that can contribute to and affect the pixel size of a desired image, such as the pixel size of the image sensor, the setup of the optical system (e.g., optical components such as an objective lens or other lenses, and the positions between the optical components within the optical system), and the way the image sensor (or more specifically the camera that includes the image sensor) is operated, such as the binning mode used or set.

[0157] As an example, the pixel size of the image sensor may be 4.8 microns. This may correspond to example (b). DDM imaging mode: image pixel size = 4.3 microns / pixel (using 4x magnification (of the objective lens) and 2x2 binning). Visual imaging mode: image pixel size = 0.86 microns / pixel (10x magnification and 1x1 binning). Here we use 4.8 microns as the pixel size of the image sensor, but other values ​​may be used. However, this may require adjusting the optical setup by using lenses with different optical properties between the objective lens and the camera (image sensor) or by changing the distance from the objective lens to the camera. Another example may be to use a camera with a pixel size of the image sensor that is 2x smaller, but records with 4x4 binning (2x larger than when using 2x2 binning). However, to reach a final size of, for example, 512x512, a camera is needed that has a 4x larger 2048x2048 image sensor, ensuring that high frame rates are still possible.

[0158] Using a 2 micron pixel size of the imaging sensor would correspond to example (a). DDM imaging mode: Image pixel size = (or approximately) 2 microns / pixel (using 4x magnification + 2x2 binning). More precisely, 1.8 microns / pixel. Calculating from the scaling, if the imaging sensor pixel is 4.8 microns, you get 4.3 microns of the image, so using a 2 micron pixel size of the sensor, it is 4.3*2 / 4.8=1.8. Visual imaging mode: Image pixel size = (or approximately) 0.4 microns / pixel (using 10x magnification and 1x1 binning).

[0159] Using an 8 micron pixel size of the imaging sensor would correspond to example (c). DDM imaging mode: Image pixel size = (or approximately) 7 microns / pixel (using 4x magnification and 2x2 binning). More precisely, 7.2 microns / pixel. Calculating from the scaling, if the imaging sensor pixel is 4.8 microns, you get 4.3 microns in the image, so using an 8 micron pixel size of the sensor, it is 4.3*7 / 4.8 / 8=7.2. Visual imaging mode: Image pixel size = (or approximately) 1.4 microns / pixel (using 10x magnification and 1x1 binning).

[0160] Although an image sensor pixel size of 4.8 microns is mentioned, it will be understood that in other embodiments there may be other values ​​that may be used (e.g., approximately 4.8 microns / pixel). For example, in some embodiments, the image sensor pixel size may be in the range of 4 to 5 microns / pixel.

[0161] It will be appreciated that in other embodiments, different magnifications of the objective lens may be used (e.g., in the range of 1x to 4x for the DDM imaging mode and 5x to 20x for the visual imaging mode), and that in other embodiments, different binning modes may be used for the DDM imaging mode and / or the visual imaging mode.

[0162] Where circumstances permit, embodiments of the invention may be implemented in hardware, firmware, software, or any combination thereof. Embodiments of the invention may also be implemented as instructions stored on a machine-readable medium that may be read and executed by one or more processors. A machine-readable medium may include any mechanism for storing or transmitting information in a form readable by a machine (e.g., a computing device). For example, a machine-readable medium may include a read-only memory (ROM), a random access memory (RAM), a magnetic storage medium, an optical storage medium, a flash memory device, an electrical, optical, acoustic or other form of propagated signal (e.g., carrier waves, infrared signals, digital signals, etc.), and the like. Furthermore, firmware, software, routines, instructions may be described herein as performing certain operations. However, it should be understood that such description is merely for convenience and that such operations may in fact result from a computing device, processor, controller, or other device executing the firmware, software, routines, instructions, etc., which in turn may cause actuators or other devices to interact with the physical world.

[0163] While specific embodiments of the invention have been described above, it will be appreciated that the invention may be practiced otherwise than as described. The above description is intended to be illustrative and not restrictive. Thus, it will be apparent to one skilled in the art that modifications may be made to the invention as described without departing from the scope of the claims set out below.

Claims

1. 1. A portable device for measuring at least one parameter of particles in a solution, comprising: A light source and a camera having an imaging sensor for generating a digital image; an optical system comprising at least one objective lens and / or a combination of objective lenses and / or lenses; Sample holder and wherein the sample comprises particles in a solution; A portable device, wherein the imaging sensor and / or camera has a frame rate greater than 100 frames / second, preferably greater than 200 frames / second, more preferably greater than 290 frames / second, even more preferably greater than 300 frames / second.

2. 2. The portable device of claim 1, wherein the frame rate greater than 100 frames per second has an image size of at least 512x512 pixels (+ / - 5%), preferably the frame rate greater than 200 frames per second has an image size of at least 300x300 pixels (+ / - 5%), or more preferably at least 328x328 pixels (+ / - 5%), and even more preferably the frame rate greater than 300 frames per second has an image size of at least 300x300 pixels (+ / - 5%), or even more preferably at least 328x328 pixels (+ / - 5%).

3. 3. The portable device of claim 1 or 2, wherein the portable device is at least partially enclosed or enclosed within a housing.

4. The portable device of claim 1 , wherein the portable device is for characterizing intensity variations in the digital image.

5. The portable device of claim 1 , wherein the at least one parameter may comprise particle mobility, preferably rate of movement, mean velocity, concentration, size, amplitude of head movement, diffusion velocity and / or frequency of head movement.

6. 2. The portable device of claim 1, wherein the optical system and / or imaging sensor is configurable or configured such that a pixel size of the digital image is at least one of, more preferably + / - 5% of and / or selectable from, the range of 0.1 micron / pixel to 10.0 micron / pixel, preferably 0.5 micron / pixel to 7 micron / pixel, more preferably greater than 2.65 micron / pixel, greater than 2.65 micron / pixel to 7.04 micron / pixel, 2 to 7 micron / pixel, 3 to 6 micron / pixel, 4 to 5 micron / pixel, 4 to 4.5 micron / pixel, 4.10 to 4.30 micron / pixel, 4.15 to 4.25 micron / pixel, 4.2 to 4.35 micron / pixel, 4.25 to 4.35 micron / pixel, and greater than 2 micron / pixel to 2.65 micron / pixel.

7. 2. The portable device of claim 1, wherein the pixel size of the digital image is substantially 0.86 microns / pixel, 0.9 microns / pixel, 1.7 microns / pixel, 2 microns / pixel, 2.1 microns / pixel, 4.3 microns / pixel and / or 7 microns / pixel, preferably + / - 5% of said values.

8. 2. The portable device of claim 1, wherein the imaging sensor has a pixel size in the range of 0.5 to 10 microns / pixel, preferably in the range of 2 to 7 microns / pixel, more preferably in the range of 2 to 5 microns / pixel, in the range of 3 to 6 microns / pixel, in the range of 4 to 5 microns / pixel, even more preferably the imaging sensor has a pixel size of 4.8 microns, even more preferably + / - 5% of said ranges or values.

9. 2. The portable device of claim 1, wherein the portable device comprises means for operating in different binning or skipping modes, preferably the imaging sensor and / or camera are configured to operate in different binning or skipping modes, more preferably configured to operate in 1x1, 2x2 and / or 4x4 binning modes.

10. 10. The portable device of claim 1, wherein the portable device comprises means for heating, cooling or maintaining the temperature of the sample and / or the sample holder at a predetermined temperature.

11. 11. The portable device of claim 10, wherein the portable device comprises a heating stage configured to heat or maintain the temperature of the sample and / or the sample holder and / or the plurality of grooves of the sample slide at the predetermined temperature.

12. 12. The portable device of claim 11, wherein the heating stage comprises a plate heated to the predetermined temperature, a resistor or resistors for heating the plate, and a temperature sensor for measuring the temperature of the plate, and preferably the portable device comprises means for maintaining contact between the heating stage and a sample slide for holding the sample.

13. The predetermined temperature may vary by + / -0.1°C, + / -0.5°C, or + / -1°C; be between ambient temperature and 50°C; be in the range of 36-41°C; be in the range of 36-41°C + / -0.1°C; be in the range of 36-41°C + / -0.5°C; be in the range of 36-41°C + / -1°C; be substantially 36°C, 36°C + / -0.1°C, 36°C + / -0.5°C, 36°C + / -1°C, 37°C, 37°C + / -0.1°C, 37°C + / -0.5°C, 37°C + / -1°C, 37.5°C, 37.5°C 11. The portable device of claim 10, wherein the temperature is at least one of 37.5°C + / - 0.1°C, 37.5°C + / - 0.5°C, 37.5°C + / - 1°C, 38°C, 38°C + / - 0.1°C, 38°C + / - 0.5°C, 38°C + / - 1°C, 39°C, 39°C + / - 0.1°C, 39°C + / - 0.5°C, 39°C + / - 1°C, 40°C, 40°C + / - 0.1°C, 40°C + / - 0.5°C, 40°C + / - 1°C, 41°C, 41°C + / - 0.1°C, 41°C + / - 0.5°C, and / or 41°C + / - 1°C.

14. 2. The portable device of claim 1, wherein the portable device comprises a processing unit for and / or configured to process the digital images.

15. 15. The portable device of claim 14, wherein the processing unit comprises at least one pre-stored processing routine for obtaining the at least one parameter, the pre-stored processing routine being configurable by a user to analyze the power spectrum of the difference between pairs of spatial Fourier transforms of digital images separated by a time delay over a range of time delays and spatial Fourier frequencies, over all possible or selected pairs of Fourier images and / or all possible or selected q values.

16. Analyzing the power spectrum of the difference between pairs of spatial Fourier transforms of digital images separated by time delays over a range of time delays and spatial Fourier frequencies (1) calculating the differential image correlation function (DICF), i.e., calculating the spatial Fourier transform of the digital images, and then calculating the power spectrum of the differences between pairs of Fourier images over all possible Fourier image pairs or subsections of Fourier image pairs, over the range of available delay times τ (i.e., the time difference between two selected images) and spatial frequencies (q) provided by the Fourier transform of the digital images; (2) then averaging all the resulting DICFs that share the same delay time (τ); (3) then radially averaging for each q value to generate the final time-averaged and vector {q}-averaged DICF as a function of delay time τ and spatial frequency q over all possible q values ​​or subsections of q values; 16. The portable device of claim 15, comprising:

17. 15. The portable device of claim 14, wherein the processing unit is for and / or configured to perform differential dynamic microscopy (DDM) over all image pairs and q values ​​or a subsection of all image pairs and / or a subsection of all q values ​​to obtain at least one parameter.

18. The portable device of claim 1 , wherein the particles are microorganisms.

19. 2. The portable device of claim 1, wherein the optical system comprises at least two objective lenses, the at least two objective lenses having at least one different optical property, preferably the at least two objective lenses having different magnifications, and more preferably one of the objective lenses for enabling images to be captured for DDM processing of particles, and another of the objective lenses for visual inspection of particles and / or for enabling images to be captured for DDM processing of particles of different sizes.

20. 10. The portable device of claim 1, wherein the objective lens has at least one of a focal length of 15 cm, a focal length of less than 15 cm, a magnification in the range of 1x to 4x, a magnification in the range of 5x to 10x, a magnification in the range of 5x to 20x, and / or a magnification in the range of 5x to 50x.

21. The portable device of claim 1 , wherein the optical system comprises means for reflecting light from the objective lens to the imaging sensor.

22. The portable device of claim 1 , wherein the portable device is configurable and / or configured such that light from the light source is transmitted to the sample holder without undergoing refraction.

23. 2. The portable device of claim 1, wherein the sample holder is configured such that one or more predetermined positions are selectable relative to a plurality of grooves in the sample slide, and preferably the sample holder is configured to slide laterally to select positions for observing one or more grooves in the sample slide.

24. A method for measuring at least one parameter of particles in a portable device, comprising: providing a sample containing particles in a solution; introducing the sample into a sample holder in the portable device; generating a digital image from an imaging sensor in a camera in the portable device; and recording the digital image at a frame rate greater than 100 frames / second, preferably greater than 200 frames / second, more preferably greater than 290 frames / second, and even more preferably greater than 300 frames / second.

25. 1. A system comprising a portable device for measuring at least one parameter of particles in a solution and a remote processing unit, the portable device comprising: A light source and a camera having an imaging sensor for generating a digital image; an optical system comprising at least one objective lens; a sample holder; means for transferring said digital image to said remote processing unit; Equipped with A system wherein the imaging sensor and / or camera has a frame rate greater than 100 frames / second, preferably greater than 200 frames / second, more preferably greater than 290 frames / second, and even more preferably greater than 300 frames / second.

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