Apparatus, system and method for measuring properties of a sample

Abstract

The device comprises an optical device for monitoring bacterial growth of an administered liquid biological sample and is provided with a sample container port for receiving a sample container having a detection chamber for containing an administered sample. The optical device comprises: a light source emitting light along an incident beam axis that intersects the detection chamber and illuminates the drug delivery sample in the detection chamber; a first photodetector receiving light scattered by bacteria in the sample; a light collection arrangement that collects light exiting the detection chamber that has been positively scattered by bacteria in the sample, directs the collected scattered light to the first photodetector, and prevents non-scattered light from reaching the first photodetector; and at least one processor measuring the intensity of the scattered light received by the first photodetector, determining a respective representative amount or concentration of the bacteria in the sample, repeating the measuring and determining steps to determine a change in the representative amount or concentration of the bacteria over time, and determining a respective sensitivity of the bacteria to a respective drug.

Description

(19) State Intellectual Property Office (12) Invention Patent Application (10) Application Publication Number (43) Application Publication Date (21) Application Number 202512028529.5 (22) Application Date 2021.02.01 (30) Priority Data 2001397.5 2020.01.31 GB (62) Divisional Application Data 202180026585.2 2021.02.01 (71) Applicant: Biological Radiation Laboratory Co., Ltd. Address: California, USA (72) Inventors: N. Wood, A. Carl, P. Harding, G. Sanders, E. Chernside, M. Malleha (74) Patent Agency: Longtian Intellectual Property Agency Co., Ltd. 72003 Patent Attorney: Huang Yan (51) Int.Cl. G01N 21 / 51 (2006.01) (54) Title of Invention: Apparatus, System, and Method for Measuring the Properties of a Sample (57) Abstract: The apparatus includes an optical device for monitoring bacterial growth in a liquid biological sample administered to an agent, and is provided with a sample container port for receiving a sample container having a detection chamber for containing the administered sample. The optical device includes: a light source emitting light along an incident beam axis intersecting with and illuminating the administered sample in the detection chamber; a first photodetector receiving light scattered by bacteria in the sample; a light collection arrangement collecting light that has been positively scattered by bacteria in the sample leaving the detection chamber, guiding the collected scattered light to the first photodetector, and preventing non-scattered light from reaching the first photodetector; and at least one processor measuring the intensity of the scattered light received by the first photodetector, determining a corresponding representative amount or concentration of bacteria in the sample, repeating the measurement and determination steps to determine the change in the representative amount or concentration of bacteria over time, and determining the corresponding sensitivity of bacteria to a corresponding drug. Claims 2 pages, Description 24 pages, Drawings 23 pages, CN 121917505 A 2026.04.24 CN 1 21 91 75 05 A 1. An apparatus comprising an optical device for monitoring bacterial growth in a liquid biological sample administered to an agent, the apparatus comprising: a sample container port for receiving a sample container in use, the sample container having at least one detection chamber for containing an administered sample; and the optical device comprising: a light source configured to emit light along an incident beam axis, the incident beam axis intersecting with at least one detection chamber of the sample container in use and illuminating the administered sample contained in the detection chamber; a first photodetector configured to receive light scattered by bacteria in the sample; and a light collecting arrangement configured to: collect light exiting the detection chamber, the light having been dispersed by bacteria in the sample relative to the incident beam axis.The device scatters light in a forward scattering angle range of approximately + / -4 degrees and + / -20 degrees, and directs the collected scattered light to the first photodetector; and prevents non-scattered light traveling parallel to the incident beam axis and leaving the detection chamber from reaching the first photodetector; and at least one processor configured to: measure the intensity of the scattered light received by the first photodetector; determine a representative amount or concentration of bacteria present in the sample based on the intensity of the scattered light; repeat the measurement and determination steps at a series of predetermined intervals to determine a representative amount or concentration of bacteria present in the sample over time; and determine the corresponding sensitivity of the bacteria in the sample to the corresponding drug. 2. The device of claim 1, wherein only light collected in an angle range of approximately + / -4 degrees and + / -20 degrees relative to the incident beam axis is directed to and received by the first photodetector. 3. The device of claim 1 or 2, wherein the range of the scattering angle of the collected scattered light is between +4 and +16 degrees and between -4 and -16 degrees relative to the incident beam axis. 4. The apparatus according to any one of the preceding claims, wherein the light collecting arrangement comprises: (1) a concave elliptical reflector configured to collect and reflect only light within an angular range of approximately + / -4 degrees and + / -20 degrees relative to the axis of the incident beam toward the first photodetector so that it is received by the first photodetector; or (2) a concave elliptical reflector configured to collect and reflect positively scattered light within an angular range of approximately + / -4 degrees and + / -20 degrees relative to the axis of the incident beam toward a concentrator, the concentrator being arranged to receive light reflected by the concave elliptical reflector and focus the received light onto the first photodetector. 5. The apparatus according to any one of the preceding claims, wherein the light collecting arrangement includes a concave elliptical reflector shaped to reflect forward-scattered light from the sample to the first photodetector or condenser, and wherein the concave elliptical reflector includes an aperture aligned with the incident beam axis and configured to allow non-scattered light from the detection chamber to pass through the concave elliptical reflector. 6. The apparatus according to any one of the preceding claims, wherein the optical device further includes a second photodetector arranged to receive non-scattered light; optionally, wherein the second photodetector is positioned on the side of the light collecting arrangement opposite the sample and aligned with the incident beam axis to receive the non-scattered light. Claims 1 / 2 pages 2 CN 121917505 A7. The apparatus of any one of claims 1 to 3, wherein the light collecting arrangement comprises a first condenser lens, a second condenser lens, and a reflector, wherein: the first condenser lens is configured to guide the scattered light to the second condenser lens and focus non-scattered light traveling along the incident beam axis onto the reflector; and the second condenser lens is configured to receive the scattered light from the first condenser lens and focus the scattered light onto the first photodetector; wherein the reflector is arranged along the incident beam axis between the first condenser lens and the second condenser lens and is configured to reflect the non-scattered light away from the first photodetector. 8. The apparatus of claim 7, wherein the reflector is arranged to reflect the non-scattered light toward the second photodetector, and the second photodetector is configured to receive the non-scattered light from the reflector. 9. A system for monitoring bacterial growth in an administered liquid biological sample, the system comprising: the means as defined in any of the preceding claims; and a sample container comprising a plurality of detection chambers, each detection chamber configured to contain the administered liquid biological sample; wherein the system further comprises: a sample positioning mechanism configured to sequentially align each of the plurality of detection chambers with the axis of the incident light beam such that the light source illuminates the administered liquid biological sample contained in the illuminated detection chamber. 10. A method for determining the drug sensitivity of bacteria in a sample, the method comprising: containing a liquid biological sample to be administered in a detection chamber of a sample container; illuminating the sample in the detection chamber with light emitted by a light source along an incident beam axis passing through the detection chamber; collecting light scattered by interaction with bacteria in the sample by a light collector, the light being positively scattered within a scattering angle range of + / -5 degrees to + / -20 degrees relative to the incident beam axis; focusing the collected scattered light onto a first photodetector by the light collector; determining, by a processor, the intensity of the scattered light collected by the first photodetector and a corresponding representative amount or concentration of bacterial particles present in the sample; repeating the determination step by the processor at a series of predetermined intervals; determining, by the processor, a representative change in the amount or concentration of bacteria in the sample over time; and determining, by the processor, the sensitivity of the bacteria in the sample to a drug for administration to the sample based on the determined representative change in the amount or concentration of bacteria in the sample over time. Claims 2 / 2 Page 3 CN 121917505 A Apparatus, System and Method for Measuring the Properties of Samples

[0001] Related Applications

[0002] This application is a Chinese patent application entitled "Apparatus, System and Method for Measuring the Properties of Samples".This is a divisional application of Chinese patent application 202180026585.2. This Chinese patent application 202180026585.2 is a Chinese patent application that entered the Chinese national phase on September 30, 2022, under PCT international application PCT / GB2021 / 050223 filed on February 1, 2021. PCT international application PCT / GB2021 / 050223 claims priority to UK patent application GB2001397.5 filed on January 31, 2020. Technical Field

[0003] This disclosure relates to apparatus, devices, systems, and methods for measuring the properties of samples, particularly for measuring the optical properties of biological samples containing microbial particles (such as bacteria). Background Art

[0004] Many techniques for measuring the properties of samples are known, particularly techniques for measuring the optical properties of biological samples (such as the concentration of particles within the sample). For example, absorption spectrophotometers measure the relative absorbance of a sample; scattering spectrophotometers, flow cytometers, and nephelometers measure the light scattered by particles in a sample.

[0005] Practical applications of this measurement technique lie in processing and analyzing clinical samples (such as urine, blood, etc.) where it is necessary to determine the number and / or type of bacteria present in a given sample to determine the most appropriate treatment modality for the patient / subject (e.g., the type and dose of antibiotic to be administered). For example, measuring the time-dependent bacterial concentration of a clinical sample that has been administered a specific drug or antibiotic allows for the determination of bacterial sensitivity to the drug (and thus the efficacy of the drug relative to a specific bacterial strain present in the sample).

[0006] Various systems using this technique are known to be available for these practical applications. For example, WO 2016 / 128747 incorporates the use of an integrating sphere light collector in which the (administered) clinical sample to be analyzed is placed. When a sample is irradiated, the bacteria present in the sample scatter some light; this scattered light is reflected and diffused by the reflective inner surface of the integrating sphere. WO 2019 / 166799 describes a similar system comprising a pair of optically connected chambers that use an integrating light collection mechanism to detect the light scattered by the sample. WO 2018 / 091922 describes a combination cartridge in which a clinical sample can be initially incubated to increase the concentration of bacteria present in the sample; subsequently, multiple independent sample portions are combined with various different drugs / antibiotics, and the optical properties of each individual sample portion are measured to determine the sensitivity of the bacteria to each drug.

[0007] It is against this background that the present device / apparatus, system, and method have been conceived.

[0008] According to one aspect of this disclosure, an optical device / apparatus is provided for monitoring bacterial growth and / or determining the number or concentration of bacteria in a liquid biological sample administered to an agent, the device / apparatus comprising: a light source configured to emit light along an incident beam axis, intersecting with a detection chamber of a sample container containing an administered sample in use, and illuminating the administered sample contained in the detection chamber; and a first photodetector configured to receive light from the sample. (See specification 1 / 24 page 4 CN 121917505 A) The light scattered by particles in this sample; a light collection arrangement configured to: collect light exiting the detection chamber, said light having been positively scattered by particles in the sample at a scattering angle between approximately + / -4 and + / -20 degrees relative to the incident beam axis, and guide the collected scattered light to a first photodetector, and prevent non-scattered light traveling parallel to the incident beam axis and exiting the detection chamber from reaching the first photodetector; and at least one processor configured to: measure the intensity of the scattered light received by the first photodetector, determine a corresponding representative amount or concentration of particles present in the sample based on the intensity of the scattered light, repeat the measurement and determination steps at a series of predetermined intervals to determine the change in the representative amount or concentration of particles present in the sample over time, and determine the corresponding change in the amount or concentration of particles in the sample. Preferably, the particles in the sample are bacteria, and the change in the representative amount or concentration of the particles over time indicates the sensitivity of the bacteria in the sample to a drug / antibiotic. In some embodiments, positively scattered light can be measured at a scattering angle between approximately + / -5 and + / -20 degrees relative to the incident beam axis.

[0009] Advantageously, the apparatus of this disclosure is capable of capturing most (e.g., about 95%) of the positively scattered light generated by the interaction of the incident beam with particles in the sample, because this light is scattered primarily at relatively small scattering angles (e.g., those mentioned above). Therefore, the apparatus of this disclosure maximizes the amount of scattered light intensity that can be collected from a single sample, which in turn means that statistically significant results can be obtained using a relatively small sample volume, particularly for changes in scattered light intensity over time.

[0010] In embodiments of this aspect of the disclosure, the amount or concentration of particles / bacteria is determined quantitatively. In other embodiments, it is suitable to determine the amount or concentration of particles / bacteria qualitatively, i.e., such that the relative amount or concentration is determined as changing over time: such qualitative measurement is appropriate and may be preferred for simplifying data processing, because changes in the amount or concentration of particles / bacteria can generally be relative to other...Reliable evaluation is performed on the time points and assays. In any embodiment of this aspect of the disclosure, the optical device is configured to receive a sample container containing at least one detection chamber for containing a biological sample. The biological sample may potentially contain pathogenic bacteria.

[0011] In some embodiments, the first photodetector may also be used in combination with a lock-in amplifier to isolate received signals having a specific frequency. The frequency to be isolated may correspond to the modulation frequency of the light emitted from the light source. This advantageously allows filtering out noise signals from other frequencies, thereby improving the signal-to-noise ratio of the obtained signal.

[0012] Suitably, the light collection arrangement is configured such that only positively scattered light within an angular range of approximately + / -4 and + / -20 degrees relative to the incident beam axis is collected and directed toward the first photodetector. More specifically, the range of the scattering angle of the collected scattered light is between +4 and +16 degrees and between -4 and -16 degrees relative to the incident beam axis, and even more specifically between +5 and +16 degrees and between -5 and -16 degrees relative to the incident beam axis. Advantageously, given that most (approximately 95%) of the light scattered by interaction with particles in the sample is scattered only at a small angle relative to the incident beam axis, the above configuration maximizes the proportion of scattered light that can be collected by the first photodetector while minimizing the amount of unscattered light that is also collected.

[0013] In some embodiments, the light collection arrangement includes: (1) a concave elliptical reflector configured to collect and reflect light only toward the first photodetector in an angular range between approximately + / -4 and + / -20 degrees relative to the incident beam axis, so that the light is received by the first photodetector; or (2) a concave elliptical reflector configured to collect and reflect positively scattered light toward a concentrator in an angular range between approximately + / -4 and + / -20 degrees relative to the incident beam axis, the concentrator being arranged to receive the light reflected by the concave elliptical reflector and focus the received light onto the first photodetector. This configuration advantageously allows the use of only one main component (or even two cooperating components) to collect light within the desired scattering angle range, thereby minimizing the number of components required in the light collection arrangement (which therefore need to be precisely positioned and aligned with each other). Furthermore, the use of a single reflector (of suitable shape and size) is beneficial because it reduces the space required to accommodate the light collection arrangement, which in turn allows for a reduction in the overall size of the device. It should also be noted that the use of a focusing element or concentrator further increases the amount and proportion of scattered light that can be collected by the photodetector, potentially improving the sensitivity of the entire device to detect small changes in the intensity of the scattered signal.

[0014] In some embodiments, the light collection arrangement includes a concave elliptical reflector shaped to reflect forward scattered light from the sample to a first photodetector or concentrator, and wherein the concave elliptical reflector includes an aperture aligned with the incident beam axis and configured to allow non-scattered light from the detection chamber to pass through the concave elliptical reflector. This configuration advantageously prevents unintentional collection of non-scattered light along with scattered light and helps ensure a representative measurement of the actual scattering (and thus a more accurate, representative determination of the amount of particles present in the sample). This arrangement, particularly in combination with the aforementioned range of scattering angles, allows for a balance between capturing as much scattered light as possible while minimizing the amount of non-scattered light also captured, while maintaining a sufficiently wide light beam to adequately illuminate the sample and produce a large amount of scattering. Furthermore, the ability to maintain constant alignment of the light beam with the aperture is advantageous for optimal light collection. Non-scattered light may be directed to a beam collector or to an exit port.

[0015] In some embodiments, the optical device further includes a second photodetector arranged to receive unscattered light; optionally, the second photodetector is positioned on the side of the light collection arrangement opposite the sample and aligned with the incident beam axis to receive the unscattered light. Advantageously, the second photodetector enables the collection and processing of unscattered (“baseline level”) light, which in turn allows the determination of the characteristics of that light. If the noise obtained in the scattered and unscattered light is common or relevant (e.g., noise due to vibrations from motor operation within the device), these characteristics can be utilized to implement noise reduction techniques during the processing of the scattered light collected by the first photodetector. Additionally or alternatively, the unscattered light can be analyzed to assess the stability of the light source and to determine variations in the intensity of the incident light that may affect the scattered light signal.

[0016] In some embodiments, the light-collecting arrangement includes a first condenser lens, a second condenser lens, and a reflector, wherein: the first condenser lens is configured to guide scattered light to the second condenser lens and focus non-scattered light traveling along the incident beam axis onto the reflector; and the second condenser lens is configured to receive scattered light from the first condenser lens and focus the scattered light onto a first photodetector. The reflector may be arranged between the first and second condenser lenses along the incident beam axis and may be configured to reflect non-scattered light away from the first photodetector. This arrangement provides an alternative configuration capable of separating scattered and non-scattered light for their respective processing and analysis, and may offer advantages over the previously described light-collecting arrangement including an elliptical reflector. For example, although more components are used in the two-condenser lens arrangement, these components may be easier to manufacture or more readily available (e.g., COTS or commercially available off-the-shelf components may be more readily available).

[0017] In some embodiments, a mirror is arranged to reflect non-scattered light toward a second photodetector configured to receive the non-scattered light from the mirror. As described with respect to configurations including elliptical reflectors, the ability to collect and process non-scattered light offers advantages in processing the corresponding scattered light signals, and particularly in terms of noise characterization and reduction.

[0018] Suitably, the device may also include a sample container turntable arranged within a sample container port for engaging a sample container and configured to align a detection chamber containing at least a portion of a biological sample within the sample container with the incident beam axis of the light source of the optical device. Advantageously, the device may be configured to engage with a separate sample container containing a sample to be irradiated by using the sample container turntable, which provides an interfacing and engagement function between the device and the sample container. This increases the ease with which one or more samples can be aligned with the optical device for measurement purposes.

[0019] In some embodiments, the apparatus further includes a motor operatively coupled to a sample container turntable and configured to rotate the turntable to periodically align and misalign a detection chamber containing at least a portion of a biological sample with the incident beam axis of the light source. Automating the alignment of a specific sample portion with the incident beam advantageously streamlines the measurement process and improves the efficiency of obtaining multiple different measurements of scattered light intensity, as well as the repeatability of such measurements.

[0020] Suitably, for example, the sample container turntable can be configured to engage with a sample container comprising multiple detection chambers, and the sample container turntable can be configured to rotate such that each of the multiple detection chambers of the sample container is sequentially aligned and misaligned with the incident beam axis of the light source. Advantageously, the above configuration provides improvements related to the parallelization of sample processing and analysis procedures: combining a rotatable sample container turntable with a sample container comprising multiple detection chambers means that multiple individual sample portions can be irradiated and scattered light intensity can be obtained in one measurement cycle / run. This increases the number of samples that can be analyzed within a given time period, which is particularly advantageous when used in the context of determining the sensitivity of bacteria in a sample to one or more different drugs (and / or drug concentrations). This, in turn, means that the appropriate drug for treating a specific patient / subject can be determined quickly and efficiently, which is especially useful when implementing point-of-care (POC) devices.

[0021] In some embodiments, the sample container turntable includes one or more openings configured such that, in use, when the sample container is properly engaged with the sample container turntable, the one or more openings interact with the sample container's...One or more detection chambers are aligned. The sample container turntable, as a whole, provides support and movement for the sample container, wherein the openings provided advantageously ensure that only appropriate / selected portions of the sample container (e.g., those corresponding to the one or more detection chambers) are positioned / placed to align with the incident beam and illuminated as needed.

[0022] In some embodiments, the sample container turntable includes one or more detectable calibration features for determining the position and / or orientation of the sample container turntable relative to the incident beam axis of the light source. The above configuration advantageously allows for the determination of when or when a given detection chamber is aligned with the incident beam, thereby ensuring that the appropriate portion or "window" of the measurement signal intensity corresponding to the signal obtained from that particular detection chamber can be extracted, processed, and analyzed. This is beneficial for reducing the effects of any drift or inconsistency in the motor rotation speed (which may occur during the measurement cycle), thereby improving the signal-to-noise ratio of the obtained signal.

[0023] Suitably, the apparatus may also include a calibration feature reader, which, in use, communicates with the processor of the apparatus to determine the time interval between the detection of the calibration feature by the calibration feature reader and the associated start of alignment of the detection chamber with the incident beam axis of the light source. The calibration reader can be configured to detect the presence of a calibration feature (e.g., through the calibration reader itself) and associate the detection of the calibration feature with the location of the associated detection chamber of the sample container.

[0024] In some embodiments, the processor of the optical device: (1) communicates with a first photodetector to measure the intensity of scattered light received by the first photodetector during a predetermined time window corresponding to a time period in which the detection chamber of the sample container is aligned with the incident beam axis of the light source; and (2) adjusts the length of the predetermined interval based on the detection of the calibration feature. Advantageously, this provides an alternative or additional mechanism for ensuring a good correlation between the presence of the detection chamber in the path of the incident beam and the extraction or processing of the appropriate portion of the obtained signal for analysis.

[0025] In some embodiments, the processor of the optical device is programmed to periodically repeat a measurement step of measuring the intensity of scattered light received by a first photodetector and a determination step of determining the corresponding representative amount or concentration of bacteria present in the sample over time, said time period being between about 20 minutes and about 2 hours, between about 20 minutes and about 1.5 hours, between about 20 minutes and about 1 hour, or between about 30 minutes and about 1 hour. This device allows the intensity of scattered light to be monitored and analyzed over a long period, thereby enabling the detection and evaluation of changes in the measured intensity over that time period. For example, see page 4 / 24 of specification 7 CN 121917505 A. Such changes can be used to determine changes in the number of particles in the sample, which indicates relevant changes in bacterial growth within the sample, and can indicate increased (or decreased) sensitivity (or lack thereof) of bacteria to a specific drug being tested.

[0026] In some embodiments, the apparatus may include a temperature control system for controlling the air temperature inside the apparatus. Suitably, the apparatus may also include at least one heating element and optionally at least one airflow regulator arranged in use to contact warm air with a sample container received within the sample container port of the optical device to maintain the biological sample in the detection chamber at a desired temperature. For example, maintaining the temperature between 36°C and 37°C is a favorable temperature relative to bacterial growth within the sample being analyzed; maintaining such a temperature during measurement cycles optimizes the conditions for bacterial growth during measurement cycles.

[0027] In some embodiments including a pair of heating elements, each heating element is operatively associated with a fan in use to push warm air toward the sample container port to heat the sample in the detection chamber of the sample container received within the device port. Such a configuration is particularly advantageous or useful for ensuring a uniform flow of heated air through and over the sample container in situ within the apparatus to maintain a desired temperature distribution on the sample container.

[0028] In some embodiments, when used for samples comprising urine from a patient or subject, the light source of the optical device is a laser light source, for example, between 620 nm and 780 nm, or in the wavelength range corresponding to visible red light. However, other wavelengths of light may also be used, particularly when the sample corresponds to other biological fluids such as blood. It should also be noted that the use of a laser light source (such as a laser diode) allows the frequency and amplitude of the emitted light to be controlled and modulated relatively easily as needed; the phase of the modulated signal can also be controlled.

[0029] In some embodiments, a predetermined interval at a rotational speed of 100 rpm corresponds to approximately 0.6 seconds. This corresponds to a suitable measurement frequency at a suitable motor speed, but it should be understood that the measurement frequency and / or motor speed may be changed as needed (e.g., a faster rotational speed of approximately 200 rpm may also be used, which would apply a greater force to the fluid sample under consideration); furthermore, the processing of the obtained signal may involve averaging multiple individual measurements, which is beneficial for noise reduction.

[0030] In some embodiments, the processor of the optical device is configured to identify multiple periodically occurring peak features in the measured light intensity and to perform measurement and determination steps only between adjacent peak features. This configuration advantageously ensures that the desired portion of the signal (corresponding to scattered light generated by interaction with the sample in the detection chamber) is extracted from the acquired signal. It can also, for example, minimize the noise level in the extracted signal by avoiding the inclusion of light scattered by other parts of the device (e.g., the sample container turntable or a portion of the sample container that does not contain any sample) in the extracted signal. This improves the signal-to-noise ratio of the signal being processed.

[0031] According to another aspect of this disclosure, a method for monitoring bacterial growth in a liquid biological sample administered with medication is provided.The system includes: the apparatus / device defined above; and a sample container comprising a plurality of detection chambers, each configured to contain a drug-administered liquid biological sample; wherein the system further includes: a sample positioning mechanism configured to sequentially align each of the plurality of detection chambers with the axis of an incident beam, such that a light source illuminates the drug-administered liquid biological sample contained in the illuminated detection chamber. As described above regarding the apparatus / device, the ability to process multiple different samples simultaneously during a measurement cycle has benefits associated with parallelization of sample processing, correspondingly increasing the speed and efficiency of obtaining useful results, and reducing associated costs.

[0032] Suitably, the sample positioning mechanism may include a rotation or turntable mechanism configured to rotate the sample container so that each of the plurality of detection chambers is sequentially aligned with the axis of an incident beam.

[0033] In some embodiments, the system may also include a support structure arranged to support the optical device. Providing such a support structure can advantageously allow the optical components to be disconnected or isolated from the rest of the system components, which, for example, reduces the impact of the operation of the sample positioning mechanism / motor on the optical components during measurement and the vibrations caused by their alignment with each other. The support structure may include an opening configured to receive a portion of a sample container comprising at least one of a plurality of detection chambers, such that when this portion of the sample container is located within the opening, at least one of the plurality of detection chambers can be positioned along the incident beam axis between the light source and the light collector. This configuration advantageously ensures good / proper alignment of the sample container (and each detection chamber disposed therein) with the incident beam. Furthermore, providing a properly configured opening in the support structure can be beneficial in guiding the user to correctly interface the sample container with the sample port when inserting the sample container into the device.

[0034] Suitably, the system may also include a temperature control system configured to maintain the temperature of the liquid biological sample between approximately 35° and 37.5°. In some embodiments, the temperature control system includes a heating arrangement comprising a heating element arranged to generate heat, and an air circulation system configured to uniformly distribute the generated heat across a plurality of detection chambers of the sample container. In some embodiments, the circulation system includes at least one recirculation duct and an associated fan arranged to drive airflow through the heating element. As described above regarding the apparatus, the arrangement maintains an appropriate desired temperature within the apparatus and around the sample container, a temperature suitable for promoting bacterial growth within the sample. This increases the concentration of bacteria in the sample, thereby increasing the corresponding intensity of scattered light obtainable from a given sample volume.

[0035] According to another aspect of this disclosure, a method for determining the susceptibility of bacteria in a sample to a drug is provided.A method comprising: containing a drugged liquid biological sample in a detection chamber of a sample container; illuminating the sample in the detection chamber with light emitted by a light source along an incident beam axis passing through the detection chamber; collecting light scattered by interaction with bacteria in the sample by a light collector, the light being positively scattered within a scattering angle range of + / -4 and + / -20 degrees relative to the incident beam axis; focusing the collected scattered light onto a first photodetector by the light collector; determining, by a processor, the intensity of the scattered light collected by the first photodetector and the corresponding degree of bacterial growth in the sample; repeating the determination step by the processor at a series of predetermined intervals; determining, by the processor, the change in the degree of bacterial growth in the sample over time; and determining, by the processor, the sensitivity of the bacteria in the sample to a drug used for drug administration to the sample based on the determined change in the degree of bacterial growth in the sample over time. In some embodiments of this disclosure and in any other aspect of this disclosure, the scattered light is detected within a scattering angle range of + / -5 and + / -20 degrees relative to the incident beam axis.

[0036] It will be appreciated that the various features and associated benefits / advantages described above with respect to the apparatus and / or system are equally applicable to the methods described above.

[0037] For example, in some embodiments, the sample container includes a plurality of detection chambers, at least two of which contain samples administered with different drugs, and the method includes: sequentially positioning each of the plurality of detection chambers containing the administered samples in light emitted along the axis of an incident beam; performing each subsequent step of the method for each of the plurality of detection chambers; and determining the relative sensitivity of bacteria in the sample to the corresponding drug used to administer the sample, to identify the most effective drug for a treatment regimen.

[0038] In some embodiments, the sample container includes a plurality of detection chambers, at least two of which contain samples administered with the same drug at different drug concentrations, and the method includes: sequentially positioning each of the plurality of detection chambers containing the administered samples in light emitted along the axis of an incident beam; performing each subsequent step of the method for each of the plurality of detection chambers; and determining the relative sensitivity of bacteria in the sample to the corresponding concentration of the drug used to administer the sample, to identify the most effective drug concentration for a treatment regimen.

[0039] Suitably, the method may further include: collecting non-scattered light parallel to the incident beam axis through said or each detection chamber by a second photodetector; and comparing the intensity of the non-scattered light collected by the second photodetector with the intensity of scattered light collected by the first photodetector for the same detection chamber. Specification 6 / 24 pages 9 CN 121917505 A

[0040] Within the scope of this application, it is expressly intended that, in the preceding paragraphs, claims and / or the following description andThe various aspects, embodiments, examples, and alternatives depicted in the accompanying drawings, especially their individual features, may be presented independently or in any combination. That is, all embodiments and / or features of any embodiment may be combined in any manner and / or combination unless these features are incompatible. The applicant reserves the right to amend any originally filed claim or accordingly file any new claim, including the right to modify any originally filed claim to subordinate to and / or incorporate any feature of any other claim, even if not initially filed in this manner.

[0041] The above and other aspects of the present disclosure will now be described by way of example only with reference to the accompanying drawings, in which:

[0042] Figures 1A and 1B show a front and rear perspective view, respectively, of an apparatus for determining the susceptibility of bacteria in a clinical sample to various drugs according to an embodiment of the present disclosure;

[0043] Figure 2 shows a vertical sectional view of the apparatus of Figure 1;

[0044] Figures 3A and 3B are perspective views of portions of the apparatus of Figure 1, and Figure 3C shows the internal airflow and temperature gradient of portions of the apparatus shown in Figures 3A and 3B;

[0045] Figure 4 is a schematic diagram of the arrangement of optical devices used in the apparatus of Figure 1 according to an embodiment of the present disclosure;

[0046] Figure 5 is a schematic diagram of an alternative arrangement of optical devices that may be used in the apparatus of Figure 1 according to another embodiment of the present disclosure;

[0047] Figure 6 is a schematic diagram of an alternative arrangement of optical devices that may be used in the apparatus of Figure 1 according to another embodiment of the present disclosure;

[0048] Figure 7 is a flowchart illustrating the various steps of a method for determining the drug susceptibility of bacteria in a clinical sample using the apparatus of Figure 1;

[0049] Figure 8 shows different time-varying detector intensity output curves in the device of Figure 1, illustrating the effects of different drugs on bacteria in clinical samples;

[0050] Figures 9A and 9B are vertical sectional views of a sample container that can be used in the device of Figure 1 to analyze clinical samples; and Figure 9C shows an exploded view of another sample container that can be used in the device of Figure 1;

[0051] Figure 10 is a plan view of the sample container of Figure 9, and Figures 10A to 10E show different parts of the flow control structure of the container of Figure 11;

[0052] Figure 11 shows a time-varying detection intensity curve measured when the sample container of Figure 10 is used in the device of Figure 1;

[0053] Figure 12A is a bottom perspective view of the sample turntable that can be used in the device of Figure 1 and intersects with the sample container of Figure 9, and Figure 12B is a perspective view of the controller used in the sample turntable of Figure 12A;

[0054] Figure 13 shows different curves of detector intensity output over time in the device of Figure 1, illustrating the signal-to-noise ratio achievable in the device of Figure 1; and

[0055] Figure 14 is a plan view of an alternative design for the sample container, which can also be used in the device of Figure 1 to analyze clinical samples.

[0056] In the accompanying drawings, the same features are indicated by the same reference numerals. Detailed Description

[0057] Specific examples and embodiments of the present disclosure will now be described, in which many features will be discussed in detail in order to provide a thorough understanding of the concepts defined in the claims. However, it will be apparent to those skilled in the art that the present disclosure (page 7 / 24, 10 CN 121917505 A) can be practiced without all specific details, and in some cases, well-known methods, techniques, and structures have not been described in detail to avoid unnecessarily obscuring the present disclosure.

[0058] FIG1 shows a front perspective view (FIG. 1A) and a rear perspective view (FIG. 1B), respectively, of device 1, which, according to embodiments of the present disclosure, can be used to provide a portable, stand-alone, and integrated module that can be used for point-of-care testing. In this case, the device can be used to test clinical samples (e.g., urine, blood, etc.) from patients or subjects to determine the sensitivity of bacteria in the samples to different types and concentrations of drugs. As described below, the device 1 is capable of rapidly (i.e., within approximately one hour) identifying and implementing particularly effective drugs and treatment regimens, thereby providing patients with effective infection treatment regimens much faster than detection and diagnostic devices relying on existing technologies.

[0059] Referring at least to Figures 1 and 2, the device detects and measures the amount or intensity of light scattered over time by particles (particularly bacteria) in a clinical sample, thereby enabling the determination of a representative amount or concentration of bacteria in the sample over time. The device 1 includes an optical device or arrangement 2 and a sample positioning mechanism 4. These two components are configured to interact with a removable sample container 6 containing the clinical sample to be analyzed in order to perform the aforementioned determination of a representative amount or concentration of bacteria.

[0060] Specifically, the sample positioning mechanism 4 is configured to engage and support the sample container 6 in such a way that at least a portion of the sample container 6 is optically coupled or connected to a component of the optical device 2. More specifically, the sample positioning mechanism 4 includes a sample turntable or sample carrier 8 and an operatively coupled motor 10, such as a BLDC (brushless DC) motor or other similar drive mechanism, which controls the movement of the sample turntable 8 (and thus the engaged sample container 6). In use, when the sample container 6 and the optical device 2 are optically coupled, the optical device 2 is configured to irradiate a portion of the clinical sample contained within the sample container 6. The optical device 2 is also configured to detect and measure the light scattered by bacterial particles in the irradiated portion of the clinical sample. The intensity of the detected scattered light can then be analyzed to determine the nature of the bacteria in the sample, particularly the relative amount or concentration of bacteria in the sample over time.

[0061] The device 1 includes a housing or enclosure 12 that houses other components of the system 2. In the illustrated embodiment, the housing 12 encloses...The device 12 includes a base 12a on which other device components are mounted; a front body portion 12b and a rear body portion 12c, which provide the walls of the housing 12; and a movable / removable cover 12d. In the illustrated embodiment, the cover 12d is hingedly attached to the rear body portion 12c; other attachment mechanisms may also be used. The cover 12d, together with the body portions 12b, 12c and the base 12a, forms a housing that accommodates the various device components when the device 1 is in use. However, it should be understood that the various portions of the housing 12 may alternatively be configured with more or fewer portions than shown herein. The device 1 also includes a closure / fastening mechanism 13 for holding the cover 12d in a closed, locked position, for example, after the sample container 6 has been inserted into the desired position within the device 1 and engaged with the sample turntable 8. The closure mechanism 13 includes an actuator 13a (shown in more detail in FIG. 2) located within the device housing 12, which is programmably actuated when the cover 12d is to be opened.

[0062] The device 1 also includes a temperature control module or arrangement 14 (highlighted in FIG. 3A) configured to maintain the temperature within the housing 12 (particularly in the area surrounding the sample container 6) within a preferred temperature range (e.g., about 36 to 38°C, more particularly about 36 to 37°C). This temperature range is particularly desirable for promoting and maintaining the growth of bacteria in clinical samples under optimal growth conditions. Furthermore, the illustrated device includes a user interface 15, such as an interactive touchscreen display, through which the user of the device 1 can interact with and program various aspects of the device 1; view certain results; and / or monitor the progress of the analysis process. For example, detailed patient or subject information can be entered by the user; measurement parameters can be displayed and changed using the interface; software updates for the device 1 can also be downloaded via user interaction with the user interface 15; measurement progress and various intermediate and final results can also be displayed to the user via the user interface 15. Furthermore, the user interface 15 can be used to provide instructions to the user to guide them through the various steps of loading the sample into the sample container 6 and subsequently properly engaging the sample container 6 with the sample turntable.

[0063] Finally, the device 1 includes one or more processors or processing units 16 that provide programmable control over various device components (e.g., optical device 2, sample positioning mechanism 4, cap closing mechanism 13, and / or user interface 15). In some embodiments, it will be appreciated that control over specific functions and components of the device can be partitioned / assigned to specific one or more of these processing units 16. In these embodiments, control over certain functions that require real-time monitoring and have relevant safety implications (e.g., related to optical device 2 and temperature control arrangement 14) can be controlled by one processing unit 16; while user interaction andControl of certain functions related to the connection (e.g., user interface 15) can be controlled by a separate processing unit 16. Furthermore, in some cases, the device 1 may be provided with side vents / openings (not shown) that can increase airflow and facilitate cooling of the processing unit 16 to prevent undesirable temperature increases within the device 1.

[0064] As described above, the sample container 6 corresponds to a separate component that can be inserted into the device 1 to enable the performance of desired measurements and tests. Generally, the sample container 6 can be provided as and / or effectively as a disposable component (i.e., usable for testing a clinical sample). Therefore, such a component is also referred to in the art as a “consumable.” Further details regarding suitable and advantageous configurations of this consumable sample container 6 will be provided subsequently with reference to Figures 9 through 11; additional details are also provided in the applicant’s co-pending application entitled “Centrifugally motivated fluidic systems, device and methods,” the contents of which are incorporated herein by reference in whole or to the extent applicable national law.

[0065] Further details regarding the configuration of the various components of the device 1 and the interactions between these components will now be provided with reference to Figures 2 and 3.

[0066] Specifically, as can be seen in these figures, the motor 10 is mounted to and supported by the base 12a of the housing 12; the motor 10 also effectively forms a support base on which many of the remaining components of the device 1 are mounted or attached. The sample turntable 8 is substantially circular in shape and is mounted above and connected to the motor 10 by a rotatable shaft 17 extending along a vertically extending axis “X” passing through the center of the sample turntable 8. Thus, rotational movement of the sample turntable 8 about the central axis “X” can be driven by the motor 10.

[0067] The sample turntable 8 includes a plurality of openings 18 arranged radially spaced around the sample turntable 8. According to the illustrated embodiment, the openings 18 are radially positioned around the outer periphery of the sample turntable 8 (these openings are shown in more detail in FIG. 12A). The sample container 6 preferably includes a corresponding plurality of detection chambers 20 (shown more clearly in FIG. 9 and FIG. 10) arranged around the sample container 6 at the same radial spacing, and each detection chamber is configured to accommodate a portion of the clinical sample to be analyzed. The openings 18 in the sample turntable 8 are arranged such that when the sample container 6 is properly joined and / or engaged with the sample turntable 8, the position of each of the plurality of openings 18 aligns with and corresponds to the position of one of the plurality of detection chambers 20 disposed within the sample container 6. Therefore, the detection chamber 20 can be located at any suitable location within the sample container 6, such as in its external region. As will be understood by those skilled in the art, the alignment between each opening 18 and the corresponding detection chamber 20 should be adapted to allow light from the light source...Light (as described below) passes through opening 18 into the corresponding detection chamber 20. This junction between the sample container 6 and the sample turntable 8 is described in more detail with reference to FIG12A.

[0068] The optical device 2 includes a light source 22 and collimating optics (not shown); a light collector or light collecting arrangement 24; and at least one photodetector 26. The light source 22 emits light along the incident beam axis “Y” and illuminates one or more sample portions present in one or more detection chambers 20 of the sample container 6. The light collector 24 collects light scattered forward by bacterial particles within the sample. Advantageously, the light collector 24 collects light scattered between angles of approximately + / -24 degrees, approximately + / -20 degrees, or approximately + / -16 degrees with respect to the incident beam axis Y. More specifically, it collects light scattered between angles of +3 degrees and +24 degrees and -3 degrees and -24 degrees; +4 degrees and +20 degrees and -4 degrees and -20 degrees; +5 degrees and +16 degrees and -5 degrees and -16 degrees on either side of the incident beam axis Y (e.g., in a ring around a specific radius of the beam). Even more specifically, the collected light can be scattered between +4 degrees and +16 degrees and -4 degrees and -16 degrees on either side of the incident beam axis Y. It should be understood that slight differences in curvature of the light collector 24 may mean that scattered light is collected at slightly different angular ranges on either side of the incident beam axis Y (e.g., between approximately 3 and 16 degrees on one side, and between approximately 4 and 16 degrees on the other). Those skilled in the art will certainly understand that light scattered at even smaller angles (i.e., less than + / - 3 or 4 degrees on either side of the incident beam axis) can also be collected; however, this would increase the proportion of unscattered incident light collected by the light collector 24. The width of the incident beam can be reduced to allow the collection of light scattered at smaller angles without including too large a proportion of unscattered light; however, this would again result in a smaller amount of sample being irradiated, thus reducing the amount of scattered light produced. Therefore, a balance needs to be struck in this respect, which will be discussed in more detail later. The collected scattered light is directed by the light collector 24 to the photodetector 26, where, for example, the intensity of the collected scattered light is analyzed to determine the relative amount or concentration of bacteria in the sample in the detection chamber at a given time point, as a result of the amount of scattered light detected. Various components of the optical device 2 are mounted onto a support plate or structure 28 to form an optical "tower," which, in the illustrated embodiment, extends substantially vertically upward from and is supported by the motor 10 or its housing 10a. However, it should be understood that the optical "tower" 28 can be mounted independently of the motor 10 and its housing 10a, or can be detached from it, to isolate the optical device 2 from any vibrations that may be generated by the motor 10. In either case, the optical tower 28 structure...Therefore, it is substantially perpendicular to the plane where the sample turntable 8 and the sample container 6 rest during use. Thus, the incident beam axis “Y” of the light emitted from the light source 22 is parallel to the rotation axis “X” of the sample turntable 8, but laterally offset by a distance “d”.

[0069] The lateral offset “d” between the rotation axis X and the incident beam axis Y substantially corresponds to the radial distance from the center of the detection chamber 20 (center) to the center of the sample container 6, and also corresponds to the radial distance between the center of the sample turntable 8 and the area of ​​the opening 18 provided in the platform. A gap or cutout 30 is provided in the support structure 28 for the optical device 2, which is located between the light source 22 and the light collecting arrangement 24 (somewhere in the vertical plane) and in the plane of the sample turntable 8; the size and position of the cutout 30 are configured to receive the radial exterior of the sample turntable 8 within it. Therefore, the receiving portion of the sample turntable 8 (and thus, the corresponding portion of the sample container 6 when it is engaged with the sample turntable 8) can extend into and through the support structure 28 and the optical tower, thereby intersecting the incident beam axis Y of the light emitted from the light source 22. In fact, the sample turntable 8, support structure 28, optical device 2, and sample container 6 are designed and adapted such that, in use, the light emitted from the light source 22 passes through one of the openings 18 of the sample turntable 8 and subsequently enters the corresponding detection chamber 20, thereby allowing the sample portion contained within the detection chamber 20 to be irradiated and analyzed.

[0070] As a result, when the sample container 6 is engaged with the sample turntable 8, each detection chamber 20 of the sample container 6 can be sequentially positioned on the incident beam axis "Y" via the optical path of the light from the light source 22 by the rotation of the motor 10 of the sample container 6. Therefore, the scattered light from the bacterial particles in the sample portion contained in each detection chamber 20 can be sequentially collected and measured by the optical device 2. In some embodiments, “measurement” refers to a quantitative assessment of the amount / intensity of light scattered by bacteria in a sample; while in other embodiments, a qualitative assessment of the relative scattering caused by samples in different sample chambers may be performed.

[0071] In the depicted embodiment, the optical device support structure 28 includes an upper (overhanging) cover portion 32 that covers some components of the optical device 2 (e.g., light collecting arrangement 24 and photodetector 26) and also provides some support for these components. The upper cover 32 also provides an additional useful function, namely, preventing unscattered light traveling along the substantially vertical incident beam axis “Y” from leaving the device 1 or accidentally reaching the user of the device 1 (e.g., in the case that the cover 12d of the housing 12 is removed when the light source 22 is emitting light). Further details regarding various configurations of the optical device 2 will then be provided with reference to Figures 4 through 7.

[0072] Turning now to the temperature control module 14, as shown in Figures 2, 3A, 3B, and 3C, this part of the device 1 is also mountedAbove and supported by the motor 10 and its housing 10a, and including at least one heating element 34 and a circulation arrangement 36. Each specification 10 / 24 pages 13 CN 121917505 A The heating element 34, such as a heating coil or arrangement / multiple heating coils, may be located within a corresponding compartment or chamber 38 positioned below the sample turntable 8. In the illustrated embodiment, a pair of heating elements 34 (see FIG. 3A) are provided in a plane below the sample turntable 8 and above the motor 10: each of the pair of heating elements 34 is located on a corresponding side of the optical device support structure 28. Those skilled in the art will understand that each heating element 34 heats the air in its vicinity during use; this heated air rises toward the underside of the sample turntable 8 and its associated sample container 6. The circulation arrangement 36 then allows the air to circulate more extensively through the device housing 12, thereby maintaining a substantially constant flow of heated air through and around the sample container 6; thus, the temperature of the contents of the sample container 6 is maintained within a desired (optimal) temperature range to promote rapid bacterial growth and proliferation. In the illustrated embodiment, the circulation arrangement 36 includes a pair of recirculation ducts 40 having an inlet 40a above the sample container 6 and an outlet 40b below the sample turntable 8. This allows for rapid adjustment of the air temperature inside the device as needed. The circulation arrangement 36 also includes one or more associated fans 42 or other mechanisms to drive airflow around the circulation arrangement 36. Specifically, in the illustrated embodiment, each heating element 34 has an associated fan 42 to drive the air heated by that heating element through the circulation arrangement 36.

[0073] FIG3C illustrates the air movement implemented by the temperature control module 14 in more detail. Each heating element-fan pair is located at the bottom of its corresponding recirculation duct 40. The fan 42 drives a constant airflow through / through the associated heating element 34; the heated air is then driven from the corresponding outlet 40b of the recirculation duct 40 into the anterior or intermediate compartment 43 below the sample turntable 8. The heated air then rises through the device, flows over and around the sample container 6, and is cooled in the process. Then, relatively cooler air enters the inlet 40a of the recirculation duct 40 to replace the air exhausted by the fan 42 and flows downward back to the heating element-fan pair. Since the recirculation ducts 40 are located on either side of the optical device 2, the material used to manufacture these ducts 40 is suitably molded and configured to prevent the heated air from causing overheating of the optical device components. This helps prevent any undesirable warping or failure of these components, or any erroneous measurements due to overheating.

[0074] It should be understood that although the devices shown in Figures 3A, 3B, and 3C have a pair of heating elements, each comprising multiple heating coils, one or more heating elements may be suitable depending on preference and design. Furthermore, although in this…In this embodiment of the invention, multiple heating coils may be advantageous in providing rapid heating of the air within the chamber, but a single heating coil may also be used, and alternatively, those skilled in the art may use many other forms and configurations of one or more heaters, and may choose according to preference. Similarly, although the device is described as having a pair of fans (one fan assigned to each heating element) to push hot air around the interior of the device, in this embodiment, fans are not necessary; for example, the airflow caused by the rotation of the sample container 6 and the sample turntable 8 can generate movement of heated air toward and around the sample container 6.

[0075] It has been shown that providing an effective mirrored arrangement of heating elements 34 and fans 42 on either side of the sample turntable 8 (each pair having its corresponding recirculation duct 40) is particularly useful for maintaining good heating balance within the device 1. It also compensates for any heating asymmetry that may arise due to the continued rotation of the sample container 6 in a particular direction. Furthermore, heating the sample container 6 in this "bottom-up" manner (i.e., air is heated and rises rapidly through the sample container 6) allows for faster and easier heating of the sample portions contained in their respective detection chambers without wasting excessive heat (and heating time) on the material of the sample container 6 itself. Therefore, in cases where the sample container 6 comprises multiple samples, each sample is located in a different detection chamber 20 surrounding the sample container 6, and the heating of the sample within each detection chamber 20 is advantageously uniform. In some cases, a desired uniform temperature of the samples in the sample container 6 can be maintained by means of a temperature measuring device (e.g., an infrared or infrared thermometer) located within the device 1 (e.g., above or near the sample container 6).

[0076] Figure 4 shows details of an example arrangement of the optical components in the optical device 2. In this arrangement, light source 22 corresponds to page 11 / 24 of the specification, CN 121917505 A, to a laser module with a laser diode, which generates light of a specific wavelength (e.g., visible red light in the wavelength range between 620 nm and 780 nm, more specifically, visible red light at approximately 635 nm) to illuminate the sample portion contained in the detection chamber 20. Note that the aforementioned wavelengths are envisioned for analyzing urine samples; however, depending on the nature of the sample to be analyzed, the wavelength of light used may differ. For example, near-infrared wavelengths (between approximately 650 nm and approximately 1350 nm) may be used for blood samples. The laser diode is connected to a signal generator (not shown) adapted to control the modulation frequency and amplitude of the laser output. Photodetector 26 corresponds to a photodiode connected to a lock-in amplifier (also not shown); the lock-in amplifier is in turn connected to the signal generator for the laser diode. This allows the photodiode to isolate and filter out signals with characteristics similar to those used for the laser diode.The specific received signal corresponds to the modulation frequency generated by the generator. This allows filtering out noise signals of other frequencies (e.g., background noise, electrical noise), thereby improving the signal-to-noise ratio obtained using optical device 2. These components can be controlled by one or more of the system's processors 16, which may take the form of one or more printed circuit boards (PCBs) or other types of microcontrollers.

[0077] In this example, the light collector 24 includes a reflector or reflective surface, which, in the illustrated embodiment, is mounted to the support structure 28 to extend through the incident beam path. Specifically, the light collector 24 in FIG. 4 corresponds to a curved concave elliptical mirror 44 with an eccentric aperture, hole, or opening 46 therein. The mirror 44 is mounted to the support structure 28 such that the aperture 46 is aligned with the incident beam axis "Y", thereby allowing unscattered light leaving the detection chamber 20 and traveling along the beam axis Y to pass directly through the mirror 44 substantially without deflection; thus preventing the unscattered light from reaching the photodetector 26. Furthermore, the reflector 44 is arranged at an angle and size such that light scattered forward by particles in the sample (particularly light scattered in the angular range of approximately +4 to +16 degrees and approximately -4 to -16 degrees along the incident beam axis Y) is reflected by the reflector 44 and toward the photodetector 26. In some other embodiments, the light reflected by the reflector 44 toward the photodetector 26 may be in the angular range of approximately +5 to +16 degrees and approximately -5 to -16 degrees along the incident beam axis Y.

[0078] Those skilled in the art will understand that when considering the sensitivity of scattered light detection, there is an interaction between the size of the light collector (reflector) aperture 46, the width of the beam from the light source 22, and the width (diameter) of each detection chamber 20 of the sample container 6 through which the beam passes. A balance needs to be struck between these values ​​to optimize the scattered light detection sensitivity. Increasing the width of the beam from the light source 22 increases the number of (bacterial) particles in the sample that are illuminated by the beam; therefore, this increases the amount of scattered light generated in any given scattering event that can be detected. However, when performing measurements of scattered light emitted from the detection chamber 20, increasing the amount of time the entire beam width illuminates the sample within the detection chamber 20 provides a longer, "cleaner" measurement path / area for the beam through the sample (away from the walls of the detection chamber 20 that may cause internal scattering, which could reduce the accuracy of detecting scattered light from the sample). Therefore, while increasing the beam width increases detection sensitivity, the beam width should still be kept smaller than the width / diameter of the detection chamber 20 to balance these two factors. Furthermore, it should be understood that configuring the system such that the diameter of the aperture 46 in the light collector (mirror) is as close as possible to the beam width will allow as much scattered light as possible to be collected and guided toward the photodetector 26, while minimizing the amount of collected non-scattered light.However, if the beam width is too close to the diameter of the aperture 46, small deviations in the path of the beam during measurement (e.g., due to component vibration or machining tolerances) may adversely affect the alignment of the beam with the aperture 46; thus, this will have a corresponding negative impact on the detection sensitivity.

[0079] In one specific embodiment, the diameter of the detection chamber 20 is envisioned to be approximately 4 mm, in which case a beam width of approximately 1 mm to 3 mm (particularly 1.5 mm to 2.4 mm) will provide a good balance between the number of irradiated bacterial particles and the length of the “clean” measurement path within the detection chamber 20. In such an embodiment, with the aperture 46 width selected to be approximately 3 mm, a beam width of, for example, approximately 1.5 mm would be suitable, as it also leaves sufficient margin between the aperture width and the beam width to account for machining tolerances and / or system vibrations. However, with the aperture 46 enlarged (e.g., to a diameter of 4 mm to match the diameter of the detection chamber 20 on page 15 of specification 12 / 24 CN 121917505 A), the beam width used can also be increased accordingly (e.g., between 2 mm and 2.4 mm).

[0080] In the arrangement of the illustrated embodiment, the concave elliptical reflector 44 reflects the positively scattered light at a defined angle, away from the incident beam axis Y (and substantially 90 degrees relative to the incident beam axis Y), to focus onto / focus the photodetector 26. One or more system processors 16 are associated with the photodetector 26 and process the detection signal generated by the photodetector 26 to calculate the intensity of the detected scattered light. A graph or plot of the detector output (which corresponds to the measured intensity of the scattered light over time) can be generated, an example of which is shown in Figure 8. The graph and / or the data used to generate the graph can be displayed to the user via the user interface 15 at regular intervals, for example, as a basic real-time indication of progress; or as a final (summary) output once the analysis process for a given sample is completed.

[0081] As shown in FIG. 4, the system of this embodiment further includes a second photodetector 48 aligned with the incident beam axis "Y", but located on the side of the light collector 24 opposite to the sample, and arranged to detect and measure unscattered light passing through the aperture 46 in the reflector 44. This second photodetector 48 also corresponds to a photodiode, configured substantially the same as the first (main) photodetector 28, i.e., the second photodetector 48 is connected to the signal generator of the light source 22 via a lock-in amplifier to ensure that the second photodetector 48 (and / or one of the processors 16 associated with the photodetector 48) is also capable of filtering out the desired laser signal frequency (and phase) from any noise signals different from the laser's frequency. Providing this additional second photodetector 48 allows for baseline measurements of the unscattered laser. This baseline measurement can be performed in conjunction with the first (main) photodetector.The intensity of scattered light measured by photodetector 26 is compared, which allows, for example, the detection of anomalies in the irradiation; laser stability can also be evaluated and taken into account during analysis. It should be understood that in any embodiment of this disclosure, the second photodetector 48 may be omitted. In some such embodiments, a beam collector or other device may be used to collect the non-scattered light from the laser.

[0082] In some cases, mirror 44 is manufactured by a custom molding process to ensure appropriate size, shape, and reflective properties. In such a process, the reflective surface of mirror 44 may be formed, for example, by coating the mirror surface with aluminum or reinforced aluminum (vapor phase). In some alternative cases, the reflectivity of the mirror (around 650 nm) should be greater than 90% (e.g., at least 95% or 98%), and the surface roughness of the mirror should be less than 100 Å (e.g., less than 80 Å, less than 60 Å, or even less than 40 Å). While the optical arrangement shown in FIG. 4 is particularly advantageous in reducing the number of components required to carry out this disclosure, other optical arrangements are also possible. For example, the custom concave elliptical mirror 44 in Figure 4 can be replaced by a pair of reflective elements, such as a first mirror deflecting forward-scattered light onto a focusing lens or a second concave mirror focusing reflected light from the first mirror onto the photodetector 26. Additionally or alternatively, one or more additional focusing components can be incorporated into the optical arrangement 2 associated with or related to the photodetector 26. For example, one or more focusing components (e.g., a focusing lens) can be arranged to form a “focusing cone” around the photodetector 26 to maximize the amount of scattered light collected by the photodetector 26.

[0083] Another optical arrangement 2' is shown in Figure 5, where an off-the-shelf concave mirror 44' can be used instead of a custom-made mirror 44' of a specific shape (but the reflectivity and surface roughness properties should be comparable). In this case, the curvature of the mirror 44' does not necessarily need to be configured to focus all (or at least substantially most) of the forward-scattered light from sample interactions onto the photodetector 26. In this case, an additional focusing element 50 (e.g., a Fresnel lens or other focusing lens) can be introduced into the optical arrangement of the optical path between the reflector 44' and the photodetector 26 to ensure that as much of the positively scattered light reflected by the reflector 44' is captured and focused onto the photodetector 26 as possible. This maximizes the total amount of scattered light available using the optical device 2', which in turn increases the overall sensitivity of the device to detect small changes in the relative number or concentration of bacteria in a sample over time. In other embodiments, as described above, the lens 50 can be replaced by a second reflector configured to reflect light onto the photodetector 26, as per specification page 13 / 24, 16 CN 121917505 A. In addition to the aforementioned focusing element 50 (or as the focusing element...),(Alternatives to the optical components), one or more additional focusing components of the optical device arrangement 2 of FIG. 4 described above can also be incorporated into optical device 2'.

[0084] FIG. 6 shows another alternative optical device arrangement 2'' that has been conceived, which can also collect scattered light and non-scattered light separately—the beam paths of the two types of light are shown in the figure. In this alternative arrangement, the light collector 24 includes a pair of focusing lenses 52a, 52b, which are spaced apart by a distance "a" (a distance greater than the focal length of the first lens 52a) and oriented such that the convex surfaces of the lenses face each other along the incident beam axis Y. In this configuration, any diverging beam incident on the first focusing lens 52a will be collimated into a parallel beam that is also parallel to the incident beam axis Y. Subsequently, such a parallel beam, when incident on the second focusing lens 52b, will converge along the incident beam axis Y toward a point at a greater distance. However, any parallel beam passing through the center of the first condenser lens 52a (or in the region corresponding to light scattered at a maximum predetermined angle from 0° to, for example, 5°) will instead be converged to a point at the focal length of the lens 52a. In the illustrated arrangement 2'', the light collector 24 also includes a reflector 54 located at a position corresponding to the focal length of the first condenser lens 52a, and its size and orientation are arranged to reflect only light arriving at the first lens at an incident angle not greater than the aforementioned maximum predetermined angle. Therefore, light passing through the sample substantially unscattered will strike the surface of the reflector before reaching the second condenser lens 52b and will be deflected away from the second lens 52b. In the depicted embodiment, the reflector 54 is angled (substantially 45 degrees) relative to the incident beam axis Y (although at least other angles may be chosen), so any beam incident on the reflector 54 is reflected at approximately 90 degrees relative to the incident beam axis Y and guided out of the main incident beam path; therefore, the beam will not reach the second condenser lens 52b.

[0085] Similar to the optical device arrangements 2, 2' shown in Figures 4 and 5, the laser diode light source 22 in the current arrangement 2'' of Figure 6 emits light along the incident beam axis Y, illuminating the sample in the detection chamber 20 of the sample container 6; bacteria in the sample scatter the incident beam in a positive direction and within a relatively small angular range. Furthermore, similar to the optical device arrangements in Figures 4 and 5, two photodetectors are provided in the optical arrangement 2'' of Figure 6: a first "off-axis" photodetector 26' is positioned at an angle (approximately 90 degrees) relative to the incident beam axis Y; and a second "on-axis" photodetector 48' is positioned aligned with the incident beam axis Y. However, in the optical arrangement of Figure 6, the non-scattered light passing through and exiting the detection chamber 20 as a parallel beam is focused onto the reflector 54 by the first condenser lens 52a, and then focused onto the offset off-axis light.On the photodetector 26'. The positively scattered beam generated by the interaction of the incident light with the bacteria in the sample is focused by a straightening lens 52a, 52b onto the on-axis photodetector 48'. This contrasts sharply with the arrangement in Figures 4 and 5, in which the off-axis (main) photodetector 26 detects the scattered light, while the on-axis photodetector 48 detects the non-scattered baseline laser.

[0086] By using one of the above-described optical device arrangements 2, 2', 2'', the device 1 is able to optimize / maximize the collection of light scattering useful in assessing the nature and / or quantity of bacteria in the sample. This is because the applicant has recognized that most (approximately 95%) of the light scattered due to such bacterial interaction is scattered only within a relatively small angular range on either side of the incident beam axis Y, for example, between angles of + / - 20 degrees to the incident beam axis Y (more specifically, between approximately 4 or 5 degrees to 16 degrees on either side of the incident beam axis Y). Therefore, instead of other forms of light collectors that require an integrating sphere light collector or utilize multiple reflections and / or diffusion of scattered light to increase collection intensity (in some cases including backscattered light), a simplified light collection arrangement can be used, which is configured to collect light scattered at those identification angle ranges most relevant to the particles in the sample being evaluated / measured, and to direct / concentrate the collected light onto a photodetector.

[0087] The method of using the above-described device 1 will now be described with reference to FIG. 7.

[0088] First, in step 105, the user securely places the clinical sample in the sample container 6, and then in step 110, inserts the sample container 6 into the appropriate position within the device 1. This includes aligning the sample container 6 with the optical tower support structure 28 (e.g., aligning the indented or cut sections or other surface features 58 of the sample container 6 with the corresponding / complementary structure of the support structure 28 or the device 1), and engaging the sample container 6 and the sample turntable 8, for example, by a snap-in / pull-out mechanism. In some embodiments, the process may be guided by a user interface 15 (e.g., via a series of diagrams and corresponding written / verbal instructions). In some embodiments, the sample turntable 8 itself may also be removable for easy cleaning as needed. The cover 12d of the device 1 is then closed and locked in place; and in step 120, the user interacts with the user interface 15 to initiate a pre-programmed sequence of actions to be taken by the various components of the device 1 to perform the desired sample analysis.

[0089] Before performing these pre-programmed actions, or actually as part of these actions, the device 1 may be configured to identify the sample container 6 in step 115 and determine information associated with that particular sample container 6 based on data provided on the sample container 6 itself or its packaging. In some cases, this information may include a unique identifier provided on the sample container 6.The unique identification code may be obtained from the sample container 6 itself, and may include a unique identifier associated with the sample container 6, a unique identifier associated with a specific batch of the sample container 6 that forms part of it, and the date of use of the contents of the sample container 6. This identification code may be provided in the form of an RFID tag or barcode (e.g., a 2D barcode), which may be scanned by device 1 before the sample container 6 is inserted (e.g., via a separate scanner associated with device 1) or even after the sample container 6 is inserted (e.g., by a scanner integrated into device 1). For example, an internal barcode scanner / reader 59 (in the device shown in FIG. 2) is shown mounted on the inner wall of the device housing 12. In this embodiment, the scanner 59 is mounted at a specific angle such that it is pointed at the identification RFID tag or barcode located on the sample container. For example, a barcode or other identification method may be located on the angled portion 58a of the sample container 6 (e.g., near the cut section 58 of the sample container so that it can be read by the scanner 59 once the sample container 6 is inserted into the device 1, as shown in Figures 9A and 9B).

[0090] Additional information may also be provided as part of or in addition to the identification code, such as details of the specific drug provided in each detection chamber 20 of a given sample container 6, so that the analysis can be performed knowing the drug being tested. Furthermore, details about software updates that may need to be implemented may also be included as part of the information provided; this allows the device 1 to easily and efficiently obtain information about appropriate software updates and changes that may need to be made from the sample container 6 itself.

[0091] Additionally or alternatively, a unique identification code may be provided on the packaging of the sample container 6 (e.g., on a box containing one or more sample containers 6 in a specific batch), or even provided with the packaging, for example in the form of a USB flash drive associated with the packaging and pre-loaded with relevant identification information. Advantageously, using packaging or a separate USB drive to provide this information increases the storage space available to hold the data, thereby enabling more data to be provided within the identification code. In this case, the device housing 12 may be provided with a port for receiving and connecting to the USB drive interface.

[0092] The device 1 may also be programmed to verify that the sample container 6 is one of the containers in an approved batch that can be used with the device 1 (e.g., to identify any counterfeit or unauthorized sample containers and prevent them from being used with the device 1). In this regard, the sample container 6 may be provided with an identification (anti-counterfeiting) feature that can be detected by the device 1; this may be provided as part of the aforementioned unique identification code or additionally. The device 1 may be programmed to reject or refuse to process any sample container 6 that does not include such a feature.

[0093] As will be described in more detail below with reference to Figures 10 and 11, the sample container 6 includes a central sample chamber 60 in which the clinical sample is initially held; and a plurality of detection chambers 20, representing the final destination of individual sample portions, in which the sample portions are irradiated and analyzed. Thus, a first series of pre-programmed actions performed by the device 1 in step 125 operates the motor 10 to rotate at a specific rotational frequency, certain predetermined directions, and a specific duration, in order to redistribute portions of the clinical sample from the central sample chamber 60 to each of the plurality of detection chambers 20. This series of actions, as described on pages 15 / 24 of CN 121917505 A, includes the following sequential steps: (a) mixing the clinical sample with growth medium 62 disposed within the sample container 6 to provide an environment conducive to bacterial growth; (b) dispensing smaller sample portions into each of a plurality of radially extending fluidic structures 64 to separate unwanted particles (e.g., precipitates, etc.) from the sample; (c) mixing the smaller sample portions with the corresponding drug (e.g., antibiotic) provided within each fluidic structure 64; and (d) retaining the final administered sample within its respective detection chamber 20 for irradiation and analysis over time. Of course, in some applications, the control sample is not exposed to antibiotics or other drugs.

[0094] Once this process has been performed and the various administered sample portions have been reassigned to their respective detection chambers 20, the next series of pre-programmed actions performed by the device 1 involves the analysis of the sample within each detection chamber 20. Motor 10 is programmed to drive the sample turntable 8 and the associated sample container 6 to rotate at a constant rotational rate (e.g., 100 or 200 rpm) over an extended period of time (e.g., over a period of approximately 60 to 90 minutes), such that a given point on the sample container 6 (e.g., a specific detection chamber 20) completes a full rotation approximately every 0.6 seconds. Therefore, in use, at a rotational speed of approximately 100 rpm or 200 rpm, each detection chamber 20 will pass through the incident beam axis Y at predetermined intervals (e.g., approximately every 0.6 seconds for a rotational speed of 100 rpm). Thus, each sample portion in its respective detection chamber 20 is sequentially (each predetermined time interval) irradiated, and the scattered light is collected by photodetector 26 and can be processed / analyzed at regular intervals during assays (e.g., bacterial growth / antibiotic susceptibility assays). Given the frequency of measurements, in some cases, it is conceivable that a (weighted) rolling average (moving average, sequential average) of the sample measurements can be used to process and combine the scattered light measurements obtained from each detection chamber 20. This will advantageously reduce the correlation with each average sample measurement point.The noise is combined (by combining the square root of the individual measurements to obtain a weighted average). For example, in some cases, it is conceivable that a smoothed average can be applied to 100 measurements (i.e., 60 seconds at a conceived rotation speed of 100 rpm). In some embodiments, the rotation speed of the sample turntable 8 (and sample container 6) is selected according to pre-programmed / factory settings; and can be determined based on the processing speed of device 1 and / or the desired measurement frequency. Thus, the rotation speed of sample container 6 during the measurement can be faster or slower than 100 rpm (e.g., between 50 rpm and 300 rpm). Similarly, the duration of the measurement can also be based on pre-programmed / factory settings, or in some embodiments can be set according to user preferences. For example, the duration of the measurement can be determined by the type of bacteria and / or the antibiotic to be tested against; and can be between about 20 minutes and 4 hours, for example between about 20 minutes and 2 hours, or between about 20 minutes and 1.5 hours. In some preferred embodiments, the duration of the measurement is between about 20 minutes and 1 hour, or between about 30 minutes and 1 hour.

[0095] As briefly mentioned earlier in this document, the signal intensity generated by the main “signal” photodiodes 26 (in optical arrangements 2 and 2') and 48 (in optical arrangement 2''), as a result of the measured scattered light intensity, is correlated with the amount and / or concentration of (bacterial) particles in the sample being analyzed / measured. In other words, a larger / stronger signal corresponds to a greater amount of light scattering, and therefore, for a greater scattered light intensity, this in turn indicates a higher concentration of (bacterial) particles in the sample. Thus, a graphical representation of the detection signal (based on scattered light intensity) over time can be used to visualize the time-varying changes in the number and / or concentration of bacteria in the sample, thereby showing and ultimately determining the sensitivity of the bacteria in the sample to the type and concentration of the drug used on that particular sample.

[0096] Figure 8 shows an example of such a graphical representation in which the sensitivity of bacteria in a clinical sample to five different antibiotic drugs was tested. In this example, the sample container 6 was divided into 28 separate detection chambers 20 and associated channels for separating the clinical sample into the 28 detection chambers 20, allowing up to 28 individual assays to be performed simultaneously. The 28 assays were divided into four zones—labeled zones 1 to 4 (see Figure 8)—and in each zone, five antibiotic susceptibility tests were performed, along with a negative control (in which the assay chamber was altered (e.g., made opaque) to prevent incident light from passing through) and a positive control (tracking uninhibited bacterial growth without any antibiotics). In this case, five different antibiotics were provided in each of the five assay chambers in each zone, as per the instructions for use on pages 16 / 24 of CN 121917505 A, allowing for four replicates per sample container 6.Bacterial susceptibility testing for each antibiotic was performed once in each of the four regions. In this way, the reproducibility of the assay around sample container 6 could also be assessed. Sample container 6 was rotated at approximately 100 rpm, and the intensity of scattered light collected from each detection chamber 20 was measured over a period of approximately 80 minutes.

[0097] As can be clearly seen from the graphs in Figure 8, during the measurement, those detection chambers used as positive controls exhibited an exponential increase in the intensity of the scattered light detected (and thus a corresponding exponential increase in bacterial number and / or concentration). This reflects the extent of the increase in bacterial number and / or concentration (under the assay conditions) that would normally be expected if no drug or other inhibitor was present and the bacteria were able to grow and replicate normally in a solution containing sufficient growth medium. Meanwhile, the detection chambers used as negative controls showed the lowest detection intensity throughout the measurement, which was also expected. Of the five detection chambers containing various antibiotics, four showed changes in detection intensity indicating a reduction in bacterial number and / or concentration (relative to the positive control) due to the action of the antibiotics; i.e., the curves had a lower or negative gradient relative to the positive control curve, but still (at least initially) had values ​​above the negative control line. Among the various administered samples, the sample that shows the greatest decrease in light scattering intensity over time compared to the positive control sample (theoretically) will correspond to the sample of the specific type and / or concentration of antibiotic most susceptible to the bacterial strain present in the administered sample. Therefore, it can be readily determined in a relatively short time which antibiotic and which concentration might be most effective in treating the patient who received the clinical sample.

[0098] Of course, it is possible (in fact, very likely) that multiple different antibiotics may be identified as antibiotics to which the bacteria in the sample may be sensitive. Therefore, various different methods have been considered for determining the most suitable antibiotic for treatment. For example, sensitivity results can be presented to the user in real time and the analysis can be terminated at any point after at least one antibiotic has been determined to be effective. However, this is not necessarily the most suitable antibiotic to administer; for example, if the response of the bacterial culture changes over a slightly longer period, or if an antibiotic initially takes longer to become effective. Alternatively, another approach could be to limit the time for performing the test (e.g., to 30 minutes, 45 minutes, or 1 hour) and present the results to the user after that time: this could mean that multiple antibiotics (or even none) and / or multiple dose levels could be considered for the patient. Another option is to only display the results after a certain number of antibiotics have been deemed effective. This, of course, means that the timescale required for testing will vary. Of course, combinations of these methods can be used.

[0099] It should be understood that testing can be based on multiple determining factors (such as the field of use of the device and / or suspected medical indications).The choice of drug or antibiotic to be used in each assay and the concentration of each drug or antibiotic is determined by the bacterial infection that may be screened. For example, when the biological sample includes urine for the determination of antibiotic susceptibility to urinary tract infections (UTIs), the antibiotic used may be selected from one or more of the following: amoxicillin; amoxicillin / clavulanic acid (2 / 1); cephalexin; ciprofloxacin; ertapenem; fosfomycin; levofloxacin; mesialin; nitrofurantoin; trimethoprim; trimethoprim / sulfamethoxazole (1 / 19). However, the selected antibiotic and its concentration or concentration range may take the form of any antibiotic (concentration) to provide the expected results during screening.

[0100] It is also evident from the comparison of the graphs of different regions that an increase in the level of signal noise was detected during rotation—the signal measurement obtained in region 4 (corresponding to the detection chamber 20 near the end of each complete rotation) showed greater noise scattering than the signal measurement obtained in region 1. The cause of the noise increase and the techniques used to address this increased noise will be described in more detail subsequently with reference to Figures 11 and 12. The results of this technique are shown in Figure 13. As can be seen from the figure, the signal-to-noise ratio variation of different detection chambers 20 in different regions of sample container 6 is much smaller and the consistency between them is higher. This will be described in more detail below with reference to Figure 13.

[0101] However, those skilled in the art will understand that the disclosed embodiments described herein provide benefits related to parallelization of sample analysis. Specifically, the design of sample container 6 means that multiple tests of different drugs (at multiple different concentrations) can be performed on samples from the same subject within a single measurement cycle, and the results can be directly compared with each other, thereby minimizing the equipment and time required to perform the evaluation. This parallel analysis of multiple different drugs and concentrations also helps to eliminate the influence of the environment on the sample test results of a single subject.

[0102] To help understand the function of the device, the consumable sample container 6 will now be briefly described in conjunction with Figures 9 and 10.

[0103] As previously described, the sample container 6 is substantially circular in its vertical cross-section and includes a common (central) sample well or reservoir 60 in fluid communication with a plurality of detection chambers 20 arranged radially outward around the container 6. Each detection chamber 20 is fluidly connected to the common sample reservoir 60 via a respective flow control structure or system 64 (including a plurality of fluid channels 66, 68). However, the detection chambers 20 are otherwise fluidly isolated from each other. The flow control structure 64, the sample reservoir 60, and the detection chambers 20 are located at the bottom of the sample container 6. The flow control structure 64 utilizes the centrifugal / centripetal force generated by the rotation of the sample container 6 by the motor 10 to cause the sample to flow from the sample reservoir 60 through the respective fluid channels 66, 68, and ultimately to the corresponding phase.Close the detection chamber 20. The radial distance between each detection chamber 20 and the central rotation axis X of the sample container 6 corresponds to “d”—the distance between the rotation axis X of the sample container 6 and the incident beam axis Y—to ensure that when the sample container 6 rotates within the device 1 in use, the beam from the light source 22 will pass sequentially through the detection chamber 20 to allow individual measurements of the scattering intensity from each sample portion in the sample container 6.

[0104] It is understood that, since the detection chamber 20 is configured to be integrated within the illustrated sample container 6, the sample container 6 (or at least the portion of the sample container 6 containing the detection chamber 20) is made of a light-transparent material (or at least a material substantially transparent to light at the wavelength of the light source) to allow light from the light source 22 to enter and exit the detection chamber 20. Specifically, in the illustrated configuration, the incident light from the light source 22 passes through the bottom or underside of the sample container 6 at a position aligned with the irradiated detection chamber 20 and exits the top surface of the sample container 6 toward the light collector 24. However, it should be understood that other configurations / relationships between the light source 22, the detection chamber 20, and the light collector 24 are also possible and fall within the scope of this disclosure.

[0105] The sample container shown in FIG10 includes 19 detection chambers 20 and their associated flow control structures 64; and has “cut-out” segments 58 with dimensions approximately equal to four or five detection chambers 20 and their associated flow control structures 64. However, it should be understood that different numbers of detection chambers and / or different sizes or shapes of cut-out segments 58 (or even no cut-out segments, as in the case of the sample container used to generate the data of FIG8) may be used without departing from the scope of this disclosure. For practicality and to maximize the efficiency of the number of flow control structures 64, for example, the cut-outs 58 may be subtend at an angle between about 20° and 60° (appropriately between about 30° and 50°).

[0106] In a convenient mode of manufacture, the flow control system 64 is formed as channels, reservoirs, and recesses in the lower surface of the sample container 6. Therefore, in order to form a closed system, a bottom cover 6c corresponding to the bottom of the sample container 6 is attached to the bottom of the sample container 6 to enclose the components and cover the flow control system 64 and sample reservoir 60 formed in the lower surface of the sample container 6 (see FIG. 9C). Suitablely, the bottom cover 6c is a membrane. Without the bottom cover 6c, the flow control system 6 would be open to the environment and would not be able to contain liquid when using the flow control device 1. The bottom cover 6c is optically transparent at least in a region axially below (some) the detection chamber 20 to allow subsequent analysis of bacteria (or other particulate matter with light scattering ability to be evaluated) in the corresponding detection chamber 20. In some embodiments, the bottom cover 6c may be opaque to light in a particularly defined region (e.g., below the detection chamber 20 which provides a negative optical control in use).

[0107] The bottom cap 6c can be attached to the bottom side of the sample container 6 using any one or more well-known suitable sealing techniques, such as heat sealing, ultrasonic welding, liquid adhesive sealing, or the bottom cap 6c may contain a single-sided / double-sided adhesive film. Regardless of the sealing technique used, it is important to ensure a high level of optical clarity and minimal reflection of incident light from the bottom cap 6c or any adhesive used (especially in the area covering the detection chamber 20) to avoid adverse effects on subsequent analysis processes. Furthermore, the sealing process should be compatible with (and should avoid interference with) any components of the sample container 6 (e.g., any drugs stored in the container). In particular, to avoid potential contamination of liquid samples in use, it is desirable that the upper surface of the bottom cap 6c within the flow control system 64 has no / minimized exposure to adhesives or other chemicals. An additional layer 6d may be attached to the bottom side of the sample container 6 (see Figure 9C) and may contain the previously described barcode or other identification method configured to be located on the inclined portion 58a of the sample container 6.

[0108] Conveniently, the sample container 6 includes a sample receiving well 69 in which an appropriate volume of sample is loaded, and a growth medium or culture medium 62 (Figures 9A, 9B) is contained axially below the receiving well 69. This medium promotes the growth of bacteria in the sample fluid and is configured to mix with the sample after it has been loaded into the receiving well 69 of the sample container 6 but before it enters the common sample reservoir 60, and is located axially below the growth medium 62. Conveniently, as in the depicted embodiment, the growth medium is dried (e.g., lyophilized or freeze-dried). However, liquid, concentrated growth medium may also be used. In alternative embodiments, powdered dried growth medium may also be used, and this can help the medium dissolve rapidly in the fluid sample. In some embodiments, it is envisioned that the growth medium may be contained in a soluble capsule / pill.

[0109] In one embodiment, the sample container 6 is provided with a cap or lid 6a that can be secured to the container 6 for use. In the depicted embodiment, the lid 6a includes an integral plunger or plug 6b and one or more protrusions 65 on the underside of the container plug 6b. When the cap 6a is engaged with the sample container 6, for example by a threaded engagement, each protrusion 65 pierces the foil / film 67 covering the growth medium 62, thereby bringing the sample into contact with the growth medium 62. When the cap 6a is further screwed into position to seal the sample within the sample container 6, the plug 69 fills at least a portion of the receiving well 69 and forces the sample and growth medium 62 into the common sample reservoir 60. It is understood that the volume of the receiving well 69 and the extent to which the plug 6b can extend into the receiving well 69 can be used to regulate the volume of sample and growth medium pushed into the common sample reservoir 60. Furthermore, the amount of growth medium contained in the sample container 6...The amount / concentration (before mixing with the sample) can be determined to provide the desired concentration of the growth medium in the volume of the sample / growth medium pushed into the common sample reservoir 60. Note that if the growth medium is contained in a soluble capsule or pill, it may not be necessary to use a foil / film 67 to cover the growth medium; therefore, the protrusion 65 for piercing the foil / film 67 can also be omitted in this case if necessary.

[0110] Once the sample and growth medium 62 have been in contact with each other and the cap 6a has been secured, the sample container 6 can be inserted into the appropriate position in the device 1. As previously mentioned, the sample container 6 is preferably designed to be inserted into the device 1 in a specific orientation. Advantageously, in the described embodiment, the asymmetry in the sample container 6, particularly the cut section 58 of the sample container 6, can be used to initially align the sample container 6 with the main optical support structure 28. Thus, in this orientation, when the sample container 6 is initially inserted into the device 1, the sample container 6 does not substantially extend through the cut 30 of the optical support structure 28. In other words, the relative alignment of the incision section 58 with the optical support structure 28 guides the insertion of the sample container 6 into the device 1 and avoids the possibility of damage to the device 1 and / or the sample container 6 due to incorrect alignment during insertion.

[0111] After the sample container 6 is inserted into the device 1, a set of programmed operations begins. The effects of these operations on the fluid sample within the sample container 6 will now be described with reference to FIG10.

[0112] As shown in FIG10A, each flow control structure 64 and its associated detection chamber 20 constitute a substantially self-contained system within the sample container 6 (indicated by circle "A" in FIG10). Although a common sample reservoir 60 exists, fluid flow between adjacent flow control structures 64 is impossible due to the centripetal / centrifugal forces in use, thereby avoiding possible cross-contamination between various drug-administered sample portions that will be analyzed separately in their respective detection chambers 20. Each flow control structure 64 includes a radially extending first “inlet” microchannel 66 (highlighted in FIG. 10A) that is in fluid communication with a clarification (precipitation) chamber or well 70 (highlighted in FIG. 10C) at its outermost radial extent. The clarification chamber 70 is in fluid communication with a generally U-shaped second microchannel 68 via a weir (dam) 70a (FIG. 10B), highlighted in FIG. 10D. This second microchannel 68 has a pair of radially extending first channel arms 68a and second channel arms 68b, configured to allow fluid to flow in a generally antiparallel direction. The first channel arm 68a connects the clarification chamber 70 to an “air spring” or pneumatic (bypass) valve 72 (sometimes referred to in the art as a “pneumatic spring” or…).The air ballast chamber extends between the two channels 66, 68a, and 68b, as highlighted in Figure 10B; while the second channel arm 68b extends between the air spring 72 and the detection chamber 20. The detection chamber 20 is highlighted in Figure 10E. Suitably, in any embodiment of this disclosure, each of the fluid channels 66, 68a, and 68b is arranged generally radially.

[0113] In this context, the term "air spring" is used to refer to an air bag that is compressed and under increased pressure as fluid (liquid) moves through the fluid channels of the flow control structure 64, and can therefore be configured to prevent fluid from flowing between the two channel arms 68a, 68b unless sufficient fluid force is applied to the gas. In other words, the air spring provides fluid sealing and inlet control functions between different areas of the flow control structure 64, such as between the two channel arms 68a, 68b. In another way, the movement of the sample fluid relative to the air spring 72 can be controlled by a balance / difference between pressures: that is, the centrifugal pressure difference caused by the rotating disk of the sample container 6 must overcome the increased pressure (overpressure) caused by the reduced gas volume before the liquid moves through the air spring valve mechanism 72. However, it should be understood that alternative valve mechanisms or sealing mechanisms may also be used to provide the corresponding function without departing from the scope of this disclosure.

[0114] First, the motor 10 rotates the sample container 6 alternately clockwise and counterclockwise (i.e., performs reciprocating / oscillating motion) at an initial relatively low speed (e.g., about 500 rpm or less) for a predetermined period of time (about 30 seconds to about a few minutes) to ensure that the sample is completely mixed with the growth medium 62 within the sample reservoir 60.

[0115] Subsequently, the sample container 6 is rotated in one direction at an increased rotational speed; the speed used varies depending on various factors (such as the geometry of the sample container 6), and thus, in some cases, speeds up to about 1600 to 1900 rpm can be used, while in other cases, where greater centrifugal force is required, speeds up to 2600 rpm can be used. This generates sufficient centrifugal force to cause the mixed sample to flow outward from the central sample reservoir 60 along each radially extending first fluid channel 66 and into the corresponding clarification chamber 70. This rotational speed is maintained for a predetermined time (e.g., about 30 seconds) to allow the sample to substantially fill the respective clarification chambers 70 and to apply sufficient centrifugal force to cause large particles (e.g., about 10 µm or larger) to deposit around the outer edge of the clarification chamber 70. Near the end of this rotational cycle, sufficient sample fluid has entered the clarification chambers 70 to raise the liquid level in these chambers above the level of the weir 70a; the overflow fluid is diverted into the first channel arm 68a of the second fluid channel 68. Any precipitates present in the sample fluid are deposited and retained in the clarification chamber 70, so that the fluid sample entering the fluid channel 68 contains essentially only particles smaller than a few micrometers (such as bacterial cells). In practice, when using...In this process, the centrifugal force acting on the liquid sample pushes heavier particles / unwanted impurities outward toward the outermost radial wall of the clarification chamber 70, thus keeping these impurities at the furthest point from the weir 70a.

[0116] Once most of the fluid sample has been redistributed from the central sample reservoir 60 to the individual flow control structures 64, the rotational speed of the sample container 6 is increased (the speed used varies depending on various factors, including the geometry of the sample container 6, and thus in some cases, the speed is, for example, above 1900 rpm or above 2600 rpm and up to about 3000 rpm, 3600 rpm or 4000 rpm). This provides sufficient centrifugal force for the sample fluid flow to overcome the pressure exerted by the air spring 72 and enter the second channel arm 68b of the second fluid channel 68. Once the "seal" (pressure block) of the air spring 72 is overcome in this way, the sample fluid is able to flow through the second microchannel arm 68b and into the detection chamber 20. The process of guiding the sample fluid from the clarification chamber 70 to the detection chamber 20 may require, for example, approximately 20 seconds, to ensure sufficient sample fluid enters each detection chamber 20 while avoiding the possibility of antibiotic contamination of the main fluid reservoir 60 (explained in more detail below).

[0117] In the illustrated embodiment, a specific type and concentration of drug (antibiotic) is present in each detection chamber 20. Conveniently, the drug / antibiotic is in a dry form, for example, due to initial deposition on and drying of the base of the detection chamber 20 during the manufacture of the sample container 6. However, those skilled in the art will understand that other forms of drug / antibiotic may be used depending on preference or suitability—for example, the drug may be deposited on paper (e.g., filter paper) placed in the area of ​​the flow control structure 64 to dissolve in the liquid sample, or the drug may be present in liquid form. Appropriately, as the sample fluid flows in and fills the detection chamber 20, it thus mixes with the deposited drug to the desired concentration. Once sufficient sample fluid has entered the detection chamber 20, the rotational speed of the sample container 6 is preferably reduced (e.g., from approximately 4000 rpm, 3600 rpm, or 3000 rpm to approximately 1500 rpm) to retain the fluid sample within the detection chamber 20. As an illustration, by reducing the rotational speed, the air in the air spring 72 expands freely again due to the lower pressure. When this occurs, the fluid in the clarification chamber 70 is pushed back into the first fluid channel 66 and into the central sample reservoir 60. This creates an air barrier between the fluids in the detection chamber 20, thus preventing cross-contamination. Furthermore, any backflow of fluid from the detection chamber 20 is prevented because once the detection chamber 20 is emptied of gas, there is no force to push the liquid sample radially inward toward the air spring 72. The fluid then passes through the sample container 6 at a relatively low speed.(For example, between approximately 1300 rpm and 1800 rpm) a predetermined period of time (approximately a few minutes, e.g., 3 minutes) of reciprocating rotation (i.e., alternating clockwise and counterclockwise rotation) is performed to inertially mix the sample fluids to solubilize their respective drugs.

[0118] The initial sample dispensing phase is now complete, and irradiation, measurement, and analysis of the samples in their respective detection chambers 20 can begin. During this process, the motor 10 is configured to rotate the sample container 6 at a relatively low speed, for example, only about 100 rpm or 200 rpm. Since this process has been described in detail above, it will not be repeated here.

[0119] Some additional details regarding the processing aspects used to extract the desired signal will now be provided with reference to FIG11, namely, the portion of the measured signal corresponding to the light scattered by the bacterial particles in the sample. The figure shows three curves of the detected signal intensity changing over time, illustrating the well-windowing technique used to extract the appropriate portion of the signal for analysis.

[0120] The inventors have noticed periodic peak features within the detection signal intensity measured by the photodetector. These features appear at points where light from the light source is incident on the walls / edges of the detection chamber 20, and have been determined to be due to increased scattering of the incident light by the inner walls of the detection chamber 20 rather than by particles within the sample. Therefore, Figure 11 illustrates how these peak features can be used during signal / data processing to more accurately determine the measured intensity signal segments corresponding to the light passing through the detection chamber 20 and scattered by the sample within the detection chamber. A (upper graphic panel) in Figure 11 depicts the scattered light intensity measured at the photodetector 26 as the sample container 6 rotates relative to the light source. The depicted scattered light trajectory spans the three detection chambers 20, i.e., the three openings 18 in the sample turntable 8 and the solid areas on either side. Removing the detection light intensity between the collection chambers 20 / openings 18 leaves only the detection scattered light intensity trajectory across the area spanned by the three detection chambers 20, as shown in the middle graphic panel (B in Figure 11). The processor can be configured to identify periodically occurring peak features in the measured light intensity (as shown in Figures 11A and 11B), which are not caused by particles in the sample, to extract the measured intensity segment between adjacent peak features from the rest of the signal (as shown in Figure 11C, lower graphic panel), so that the scattered light intensity can be measured and determined only relative to the extracted segment (corresponding to light scattering from the sample rather than light scattering from the edge of the detection chamber 20) to determine bacterial growth. This can advantageously avoid processing the portion of the signal containing large noise spikes and can allow the signal to be appropriately amplified.

[0121] Compared to simply using a predefined “time window” to extract the “desired” signal for measurement (relative to...A more direct method, namely, extracting data for each sample at a given time offset from the start of each new rotation of sample container 6 within a predefined time period or "window," offers improvements in noise reduction. This is because minute changes in motor speed cause the predefined "time window" to become inconsistent with the actual signal corresponding to the light scattered from the bacteria in the sample; that is, the predefined "time window" will "drift" relative to the actual desired signal over time. This, in turn, means that the "time window" used for data extraction will overlap with the peak features (as described above, generated by scattering from the edges of detection chamber 20), and the extracted signal will include a greater amount of this increased scattering from the edges of detection chamber 20, thus exhibiting a higher noise level. This "drift" or misalignment between the extraction "time window" and the actual time period in which the desired signal is located is more pronounced in those samples measured near the end of the rotation, as shown by the increased noise in the graph of region 4 (as shown in Figure 8). However, it should be noted that the aforementioned well-window technique may be more readily implemented in the context of post-processing of sample measurements rather than as a mechanism for real-time extraction of specific signal segments for processing. Additional or alternative mechanisms for reducing noise in the extracted signal have also been developed, which are capable of more accurately identifying the “window” where the desired signal from each detection chamber 20 is located; this mechanism will now be described in more detail with reference to Figures 12A and 12B.

[0122] Figure 12A provides a bottom perspective view of the sample turntable 8, to which further details of the calibration aspects of the intensity measurement process will now be described. Figure 12B provides a close-up view of the controller used in conjunction with the sample turntable 8 during the calibration process.

[0123] In the embodiment shown in Figure 12A, the sample turntable 8 includes an additional calibration ring or gear 80 located below it and comprising a plurality of calibration features or teeth 82. In the illustrated embodiment, the calibration features 82 correspond to a plurality of radially extending spokes projecting from the calibration ring 80 at intervals and are arranged such that each calibration feature 82 is associated with a corresponding one of a plurality of openings 18.

[0124] In use, when the sample container 6 is engaged with the sample turntable 8, each calibration feature 82 will also be associated with a corresponding one of the plurality of detection chambers 20 in the sample container 6. The device 1 also includes a calibration reader or optical encoder 84, which is arranged within the device 1 to be positioned adjacent to the underside of the sample turntable 8 and configured to sequentially intersect with each calibration feature 82 in use as the sample turntable 8 is rotated by the drive shaft 17. Specifically, the calibration reader 84 includes an optical arrangement configured to detect each calibration feature 82 passing through it, for example by detecting a reduction or loss of optical signal caused by the passage of the calibration feature 82 and temporary interruption of the beam path within the calibration reader 84 (as shown in more detail in FIG12B).

[0125] Since each calibration feature 82 is associated with one of the detection chambers 20, the calibration reader 84 can be used to detect each calibration feature 82 associated with each opening 18 of the sample turntable 8 and send a signal to the controller / processor of the device 1 to begin measuring the intensity of scattered light at a predetermined time period after the detection of the calibration feature 82 and to continue for a predetermined time period sufficient to cover the time period during which the detection chamber 20 intercepts light from the light source 22 so that it does not strike the wall of the detection chamber 20 (i.e., a time period sufficient to obtain the reading illustrated in C of FIG. 11). Advantageously, in this way, the light scattering measurement window is reset multiple times in each rotation of the sample container 6 to ensure that the photodetector reading is properly in phase with the detection chamber 20. It should be understood that the number of calibration features can be selected according to preference: for example, there may be a calibration feature associated with each opening 18 in the sample turntable 8, or there may be one calibration feature associated with a predetermined group of openings 18 (e.g., one calibration feature 82 for every 2, 3, 4, 5 or 6 openings 18, etc.).

[0126] Alternatively, the calibration feature 82' may take different forms. For example, as shown in the embodiments illustrated in Figures 2, 3A, and 3B, calibration features 82' take the form of ribs, fins, or flags that are spaced apart around the circumference of the sample turntable 8 and (as shown in these figures) extend substantially downward from the bottom of the sample turntable 8. Thus, these ribs 82' can be molded and form an integral part of the sample turntable 8. In this case, the calibration reader 84 can be instead mounted and oriented such that the vertically extending ribs 82' pass through the optical arrangement of the calibration reader 84. In the embodiments shown in Figures 3A and 3B, a calibration feature 82' is associated with each opening 18 of the sample turntable 8, such that each calibration feature 82, through a channel in the optical arrangement of the calibration reader 84, can form a trigger for reading or measuring scattered light obtained from each corresponding detection chamber 20. Advantageously, this helps prevent drift occurring in the "window" due to variations in motor speed, since a particular indicator is associated with each opening 18, and therefore with each detection chamber 20.

[0127] Figure 13 shows the detector intensity output over time in the device of Figure 1, illustrating the signal-to-noise ratio achievable using calibration features 82, 82' and calibration reader 84. As shown in Figure 8, device 1 is divided into four regions or quadrants, and each of the four graphs in the specification (pages 22 / 24, CN 121917505 A) represents a detector intensity measurement obtained from the detection chamber 20 located in the corresponding region. Note that no antibiotics or other drugs were provided in the device when the detector intensity output graphs of Figure 13 were generated, therefore (as expected) these graphs do not show any significant decrease in detector intensity output, whereas bacteria are present in the sample.If the target is sensitive to antibiotics, a significant reduction in detector intensity output can be expected (as shown in Figure 8). However, it is clear from the graph in Figure 13 that the observed signal-to-noise ratio variance is significantly reduced in all cases: the plotted detector intensity is relatively consistent across all four regions, with no region showing higher noise than any other region (as shown in region 4 of Figure 8).

[0128] In some cases, it is conceivable that the calibration reader 84 (or the processor associated with the photodetector 26) can be configured to calculate time intervals between adjacent calibration features 82 passing through the reader 84 and compare these calculated intervals with predetermined intervals for measuring the intensity of the collected scattered light. If there is a difference between the measured “calibration” time interval and the predetermined measurement time interval, and the difference exceeds the predetermined time interval, the processor can be configured to change the measurement time interval to align it with the “calibration” time interval. This ensures that intensity measurements are performed when the detection chamber 20 is precisely aligned with the axis of the incident beam, i.e., when the light from the light source is incident substantially through the center of the detection chamber 20.

[0129] Numerous modifications can be made to the above examples without departing from the scope of this disclosure as defined by the appended claims.

[0130] For example, the drug does not need to be located within the detection chamber 20, but can be located at different parts of the flow control structure 64, such as within the second channel arm 68b of the second microchannel 68. Additionally or alternatively, a second pneumatic valve or air spring can be located in the flow path after the detection chamber to implement different mechanisms for effective mixing of the sample and drug (via a back-and-forth "shaking" motion between pairs of air springs).

[0131] Furthermore, it should be noted that the design of the sample container 6 can be modified to change the optical path length of light passing through the detection chamber 20 by varying the depth of the detection chamber well. Increasing the optical path length will increase the signal: the light will pass through more sample and interact with more bacterial particles in the process. Example path lengths that can be considered are between 4 mm and 10 mm (e.g., well depths of 4 mm, 6 mm, 8 mm, or 10 mm); changing the optical path length will also involve changing the dimensions of other features in the flow control system 64, such as the clarification chamber 70 and the air spring 72.

[0132] Other mechanisms for improving the signal-to-noise ratio include “shielding” the edges of the detection chamber 20, for example by securing a thin film or plastic sheet or other thin material to the bottom of the sample container 6 to prevent light from entering the sample container 6 or interacting with other parts of the sample container—for example, the diameter of the opening 18 in the sample turntable 8 may be smaller than the diameter of the detection chamber 20.

[0133] Furthermore, detection sensitivity can be increased by making some slight (optional) changes to the optical arrangement 2 to increase the signal-to-noise ratio of the detected scattered light. For example, a second photodetector 48 that collects non-scattered light may be used relative to the non-scattered light.The beam path is tilted or angled to reduce unwanted reflections of the non-scattered light back into the optical system (where it can interact with other beams). In some cases, the first photodetector 26 can be shielded to reduce unwanted detection of stray light within the optical system.

[0134] As will be understood, minimizing the number of (individual) machined parts of the optical components when assembling them can improve the accuracy of the beam output from the light source 22 and the alignment of that beam with the aperture 46 in the detection chamber 20 and the light collector 24. Furthermore, in some cases, particularly where the light source is in the form of a laser (diode), the laser can be included in a machined / easily replicated laser block to increase the consistency of the laser beam position during measurement (and thus increase the ease of maintaining good alignment within the optical system).

[0135] Regarding maintaining consistent alignment of the beam through the optical system, additional sensors (e.g., accelerometers) may be incorporated into the device 1 to measure vibrations occurring within the device 1 (e.g., due to the rotation of fan 42 on pages 23 / 24 of the specification 26 CN 121917505 A of the motor 10, which controls the sample container 6 and / or temperature control module 14). This helps ensure that any vibrations are kept within acceptable limits / below, thereby avoiding adverse effects on the alignment of components in the optical arrangement 2.

[0136] Finally, an alternative design for the sample container 6' is also envisioned, as shown in FIG. 14. As can be seen from this figure, the relative positions of the clarification chamber 70' and the detection chamber 20' in this alternative sample container 6' differ from the relative positions of the clarification chamber 70 and the detection chamber 20 in the sample container 6 shown in FIG. 10. Specifically, in the sample container 6 of FIG. 10, the radially outermost extent of the clarification chamber 70 is located at a greater distance from the central axis of rotation of the sample container 6 compared to the radially outermost extent of the detection chamber 20. However, in the alternative sample container 6', the opposite configuration is used: the outermost radial extent of the detection chamber 20 is located at a greater distance from the central axis of rotation of the sample container 6' than the outermost radial extent of the clarification chamber 70'. The latter arrangement, shown in Figure 14, advantageously increases the ease with which the fluid sample is held within the detection chamber 20' for illumination and measurement, as the centrifugal force acting on the fluid tends to move the fluid toward the outermost radial extent of the flow control system 64. It should be understood that this adjustment of the relative position of the detection chamber 20 may require some corresponding changes to the relative positions of the components of the optical arrangement 2 to maintain the alignment of the light source 22, the detection chamber 20, the light collector aperture 46, and the photodetectors 26, 48. [Instruction manual 24 / 24 pages 27 CN 121917505 A Figure 1A Instruction manual Figure 1 / 23 pages 28 CN 121917505 A Figure 1B Instruction manual Figure 2 / 23 pages 29 CN 121917505 A Figure 2 Instruction manual]Figure 3 / 23, page 30, CN 121917505 A, Figure 3A; Figure 4 / 23, page 31, CN 121917505 A, Figure 3B; Figure 5 / 23, page 32, CN 121917505 A, Figure 3C; Figure 6 / 23, page 33, CN 121917505 A, Figure 4; Figure 7 / 23, page 34, CN 121917505 A, Figure 5; Figure 8 / 23, page 35, CN 121917505 A, Figure 6; Figure 9 / 23, page 36, CN 121917505 A, Figure 7; Figure 10 / 23, page 37, CN 121917505 A, Figure 8; Figure 11 / 23, page 38, CN 121917505 A, Figure 9A, Figure 9B; Figure 12 / 23, page 39, CN 121917505 Figure 9C, Appendix 13 / 23, Page 40, CN 121917505; Figure 10, Appendix 14 / 23, Page 41, CN 121917505; Figure 10A, Appendix 15 / 23, Page 42, CN 121917505; Figure 10B, Appendix 16 / 23, Page 43, CN 121917505; Figure 10C, Appendix 17 / 23, Page 44, CN 121917505; Figure 10D, Figure 10E, Appendix 18 / 23, Page 45, CN 121917505; Figure 11, Appendix 19 / 23, Page 46, CN 121917505; Figure 12A, Appendix 20 / 23, Page 47, CN 121917505; Figure 12B, Appendix 21 / 23, Page 48, CN 121917505 Figure 13 Appendix to the Instruction Manual, Page 22 / 23, 49 CN 121917505 A Figure 14 Appendix to the Instruction Manual, Page 23 / 23, 50 CN 121917505 A Abstract: The apparatus includes optical devices for monitoring bacterial growth in a drug-doped liquid biological sample and a sample container.port for receiving a sample container having a detection chamber for containing the administered sample. The optical singular apparatus include: a light source emitting light along an incident beam axis intersecting the detection chamber and illuminating the administered sample in the detection chamber; a first photodetector receiving light scattered by bacteria in the sample; a light collection arrangement collecting light existing the detection chamber that has been scattered forward by bacteria in the sample, guiding the collected scattered light to the first photodetector, and preventing non- scattered light from reaching the first photodetector; and at least one processor measuring the intensity of the scattered light received by the first photodetector, determining a corresponding representative amount or concentration of bacteria in the sample, repeating the measurement and determination steps to determine changes in the representative amount or concentration of bacteria overtime, and determining the corresponding susceptibility (it is used when affected by an external factor like drugs) of the bacteria to the corresponding drug.

Claims

1. An apparatus comprising an optical device for monitoring bacterial growth in a liquid biological sample administered with an agent, the apparatus comprising: A sample container port for receiving a sample container in use, the sample container having at least one detection chamber for containing a drug administration sample; as well as The optical device includes: A light source configured to emit light along an incident beam axis, wherein, in use, the incident beam axis intersects with at least one detection chamber of the sample container and illuminates a drug-eluting sample contained within the detection chamber. A first photodetector is configured to receive light scattered by bacteria in the sample; The light collection arrangement is configured as follows: The light exiting the detection chamber is collected, the light having been positively scattered by bacteria in the sample within a scattering angle range of approximately + / -4 degrees and + / -20 degrees relative to the axis of the incident beam, and the collected scattered light is guided to the first photodetector; and To prevent non-scattered light traveling parallel to the incident beam axis and leaving the detection chamber from reaching the first photodetector; and At least one processor is configured as follows: Measure the intensity of the scattered light received by the first photodetector; The representative amount or concentration of bacteria present in the sample is determined based on the intensity of the scattered light. The measurement and determination steps are repeated at a series of predetermined intervals to determine the representative amount or concentration of bacteria present in the sample over time; and Determine the sensitivity of the bacteria in the sample to the corresponding drug.

2. The apparatus according to claim 1, wherein, Only light collected within an angle range of approximately + / -4 degrees and + / -20 degrees relative to the axis of the incident beam is guided to and received by the first photodetector.

3. The apparatus according to claim 1 or 2, wherein, The range of the scattering angle of the collected scattered light is between +4 and +16 degrees and between -4 and -16 degrees relative to the axis of the incident beam.

4. The apparatus according to any one of the preceding claims, wherein the light collection arrangement comprises: (1) A concave elliptical reflector configured to collect and reflect light only toward the first photodetector in an angular range of approximately + / -4 degrees and + / -20 degrees relative to the axis of the incident beam, so that it may be received by the first photodetector; or (2) A concave elliptical reflector configured to collect and reflect positively scattered light in an angular range of approximately + / -4 degrees and + / -20 degrees relative to the axis of the incident beam toward a concentrator, the concentrator being arranged to receive the light reflected by the concave elliptical reflector and focus the received light onto the first photodetector.

5. The apparatus according to any one of the preceding claims, wherein, The light collection arrangement includes a concave elliptical reflector shaped to reflect forward scattered light from the sample to the first photodetector or condenser, and wherein the concave elliptical reflector includes an aperture aligned with the incident beam axis and configured to allow non-scattered light from the detection chamber to pass through the concave elliptical reflector.

6. The apparatus according to any one of the preceding claims, wherein, The optical device further includes a second photodetector arranged to receive non-scattered light; optionally, the second photodetector is positioned on the side of the light collection arrangement opposite to the sample and aligned with the axis of the incident beam to receive the non-scattered light.

7. The apparatus according to any one of claims 1 to 3, wherein, The light-collecting arrangement includes a first condensing lens, a second condensing lens, and a reflector, wherein: The first condenser lens is configured to guide the scattered light to the second condenser lens and focus the unscattered light traveling along the incident beam axis onto the reflector; and The second condenser lens is configured to receive scattered light from the first condenser lens and focus the scattered light onto the first photodetector; The reflector is arranged between the first condenser lens and the second condenser lens along the axis of the incident beam, and is configured to reflect the non-scattered light away from the first photodetector.

8. The apparatus according to claim 7, wherein, The reflector is arranged to reflect the non-scattered light toward a second photodetector, which is configured to receive the non-scattered light from the reflector.

9. A system for monitoring bacterial growth in a liquid biological sample after administration of an oral medication, the system comprising: The apparatus as defined in any of the preceding claims; as well as A sample container comprising multiple detection chambers, each configured to contain a liquid biological sample to be administered; The system also includes: A sample positioning mechanism is configured to align each of the plurality of detection chambers sequentially with the axis of the incident light beam, such that the light source illuminates the drug-administered liquid biological sample contained within the irradiated detection chamber.

10. A method for determining the drug susceptibility of bacteria in a sample, the method comprising: The liquid biological sample to be administered is contained in the detection chamber of the sample container; The sample inside the detection chamber is illuminated by a light source using light emitted along the axis of the incident beam passing through the detection chamber; The light scattered by interaction with bacteria in the sample is collected by a light collector, the light being scattered in the positive direction within a scattering angle range of + / -5 degrees to + / -20 degrees relative to the axis of the incident beam; The light collector focuses the collected scattered light onto the first photodetector; The processor determines the intensity of the scattered light collected by the first photodetector, and the corresponding representative amount or concentration of bacterial particles present in the sample; The processor repeats the determination steps at a series of predetermined intervals; The processor determines a representative change in the amount or concentration of bacteria in the sample over time. as well as The processor determines the sensitivity of the bacteria in the sample to a drug used to administer the sample based on a representative change in the amount or concentration of bacteria in the sample over time.

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