WO2023105263A1

WO2023105263A1 - Automatic hybrid machine vision measurement system for microbial and bacterial contamination, identification of cell count and particle homogeneous dispersion system based on mcfarland scale

Info

Publication number: WO2023105263A1
Application number: PCT/IB2021/061383
Authority: WO
Inventors: Sayedali MOUSAVI
Original assignee: Mousavi Sayedali
Priority date: 2021-12-07
Filing date: 2021-12-07
Publication date: 2023-06-15

Abstract

The Present invention device is used in chemical, microbiological, and cellular laboratories, detecting surface contaminants in many industries such as the food industry. This device is related to the McFarland scale, a reference for measuring the turbidity of microbial suspensions. This device can determine the amount of turbidity of the solution and then count the bacteria and cells in the suspension as cell and bacterial culture testing device. This invention is related to the chemistry and biomaterial section of the hybrid technology section. This hybrid device also can detect homogenies suspensions and purity of mixtures. This invention is including a uniform light source operative to emit a beam of light, a camera for sending pictures for image processing for analyzing samples which detects the intensity of pictures after the light interacts with the samples.

Description

[0002] [Automatic hybrid Machine vision measurement system for microbial and bacterial contamination, identification of cell count and particle homogeneous dispersion system based on McFarland scale.

[0003] This invention relates to a device for automatic and portable microbial contamination measurement and identification of cell count and particle homogenous dispersion system based on McFarland scale. This device has a user-friendly interface that is controlled by a mobile application.

[0004] Cross References to Related Applications: This application claims priority from Provisional Application System, AUPO2363A, US6002477A, PCT/AU1997/000603, CA2753161 C.

Filed of Invention

[0005] The present invention relates to experimental and laboratory instrumental device and equipment. The present invention relates to diagnose, Bactria, cells numbers with McFarland ration with image processing and also detect homogenous grade of suspension for detection of micro and Nano particle desperation in mixture solution and detection methods thereof.

Summary of Invention

[0006] The present invention is used to measure the concentration of bacteria or cells in a solution of bacterial and cell suspension. The McFarland method is the standard method for this type of measurement. Each degree on this scale is equivalent to 300 million bacteria in solution. Each of these degrees can be measured by attributing the turbidity of the solution. The McFarland Index is measured using the image processing approach. Relevant images are captured by a digital camera and analyzed using a Raspberry Pi processor, and finally, the turbidity of the solution is measured. This information is sent via Bluetooth to the mobile application and stored on the server. This application also can control the device. In this device, LED light with features such as biocompatibility, low energy consumption, stable light, easy interchangeability, and cost-effectiveness, has replaced ultraviolet light.

Technical Problem

[0007] The main problem in measuring standard bacterial concentrations is the lack of access to spectrophotometric devices due to their high cost. The existing devices for measuring commercialized bacteria concentration are also costly and are accompanied by problems such as low relative accuracy, low linear measurement range, and low detection limit. The food industry centers, medical diagnostic laboratories, microbiology, and cell laboratories need an inexpensive device. On the other hand, due to the dangerousness of the bacteria, the bacterial counting system in the solution requires unique settings. The purpose of designing and manufacturing this device is to reduce production costs and increase accuracy, detection limit, measuring speed, and ease of use. Moreover this hybrid detection system can distinguish dispersion of all micro and Nano materials in solvents.

Description of previous knowledge status and a history of advances related to the invention

[0008] In microbiology, clinical, and other similar laboratories, the number of bacterial cells in a culture medium or liquid medium needs to be estimated or determined [1 ]. The McFarland standard is commonly used as a reference for adjusting the turbidity of microbial suspensions (especially bacterial suspensions) so that the number of microorganisms in the culture medium used is within a specific range, and the microbial test is standard [2],

[0009] The turbidity of microbial suspensions is usually determined by a nephelometer, densitometer, or a visual comparison tool [1 ]. These methods are based on the physical principles of light scattering, which results from the interaction of light with particles in a suspension system. The turbidity of the samples affects the transmission and scattering of light and makes it possible to measure the intensity of light transmitted through the sample.

[0010] In an invention numbered US4073623A in 1978 by Robert M. Mylan White, a representative of Mylan White, unveiled a device to measure both light and dark color solution. In this device, a control tube and a sample tube, and a calibrated color table have been used as a reference for analyzing the color differences in the tested sample and the reference sample. This method is an ocular method for comparing the color of samples with low accuracy to analyze the turbidity of the solution [4],

[001 1 ] The following invention was unveiled with the number US6359689B1 , in 2002. A device called a programmable automatic colorimeter by Richard A. Rosenberger was registered. The device includes a motor for rotating the rotating plate, an aluminum plate with holes for tubes, a light source, and a detector. They are arranged so that the tubes on the rotating plate are alternately placed between the light source and the detector. An amplifier and a recording system were also used to record the turbidity of each sample. In this invention, a number with a McFarland unit is not presented, bacteria are not counted, and the device is used only as a solution colorimeter [5].

[0012] In this invention, a device was introduced as an optical densitometer. The system consists of two disks (one in the form of rotating blades and a full disk), a light source, a counter, a transmitter, and a light pulse receiver. As the light passes through the solution, the counter records a pulse. This system has not been used exclusively to investigate the turbidity of the solution [6].

[0013] In 201 1 , Mr. Zamora et al. presented a paper entitled Using Digital Photography to Implement the McFarland Method. The McFarland method allows the concentration of bacterial cells in the liquid medium to be determined by instrumental methods: turbidimetry or nephelometry.

Microbes act by absorbing and scattering event light, so the amount of absorption (turbidimetry) or intensity of light (nephelometry) measured is directly proportional to their concentration in the environment. In this work, a new analytical imaging method is developed to determine the concentration of bacterial cells in the liquid medium. Digital imagery is a set of McFarland standards used to assign turbidity-based color values using proprietary software. These values are proportional to the bacterial concentrations that allow a calibration curve to be easily constructed. This paper evaluates the reproducibility of an in vitro calibration study and compares the turbidity and nephelometric results with the results presented by the proposed method. This method is relatively simple and cost-effective. The device used in this paper is in the range of 0.5 to 4 McFarland units. New UV has also been used as a light source [7],

Solution to Problem

[0014] Conventional spectrophotometric devices are associated with long preparation and response times due to the use of sodium light sources as well as photodiode array detectors. The use of these resources also greatly increases the cost of producing devices that include these parts. In addition, many spectrophotometers cannot measure the turbidity of a solution.

[0015] The device introduced in this invention uses the white light source of light-emitting diodes and the camera as a detector. These parts have less cost and access problems compared to the examples mentioned in other devices.

[0016] Dedicated available devices for measuring the turbidity of commercially available solutions are designed and manufactured in such a way that they are not suitable for measuring the turbidity of solutions containing bacteria, especially pathogenic and dangerous species.

[0017] In this invention, a special device for measuring solutions containing bacteria or cells has been introduced. Components associated with solutions also have the ability to disinfect.

[0018] Existing special commercial McFarland-scale measurement devices are also associated with problems such as the use of expensive light sources and detection, low measurement range, measurement error and, so on.

[0019] To achieve the specified goals, the test tubes are placed in the designed hole. A homogeneous light is emitted to the test tube and the McFarland Index is sensed using a camera. This camera is focused on the tube. The device has several holes for storing test specimens (primary holes) and reservoir holes (secondary holes). The device LCD is located on the front. This monitor is used to screen the results and determine the test parameters.

[0020] All the results and collected information for all interventional in experiments from device will send to Cloud and can be accessed through device’s application for inspectors, clinician or other users .

Explanation of shapes, maps, and diagrams

[0021] As you can see in Figures A and B 1 , the, present invention device is made of the following components (The mentioned numbers correspond to the numbering of the components on the figure.1 :

1 . The bottom frame of the device: It is made of polylactic acid. The frame contains the general and internal parts of the device. 2. The front door of the device: It is completely closed by four screws designed at the bottom of the device.

3. LCD frame: It is a place to put the device LCD.

4. Camera holder: Designed with an angle control holder mechanism.

5. Mini Camera

6. Pipe fittings: Designed to hold the pipes in place and vertically.

7. LED lamp: This lamp is placed in the detection stage for exposure.

8. Heat sink: Prevents the inside of the device from overheating due to the LED lights on.

9. LCD device with dimensions of 5 inches

10. Raspberry Pi board to control the device

11 . Input port for connecting the device

12. Test tube holder: With this holder, the pipes are placed completely vertically.

13. Status display white LEDs

14. M8 screw to close the lid of the device

15. M5 screw to fasten parts including LEDs and holders

16. Transparent Plexiglas plates: These plates are used to absorb light received from the LEDs and to distribute the light evenly at the location of the tubes.

[0022] Figure 2. Shows the final design of the present invention body

[0023] Figure 3. Shows the rendered picture of the final tree dimensional design of the device body

[0024] Figure 4. Shows the Top-side of the 3D-printed present invention body

[0025] Figure 5. Shows Front view of the 3D-printed present invention body

[0026] Figure 6. Shows Tubes holder design and relative scales

[0027] Figure 7. shows the background image and added guidelines (empty holders) and chamber holes

[0028] Figure 8. Shows the Images of different McFarland standard solutions and corresponding measurements

[0029] Figure 9. Shows The main interface of present invention present invention ’s application

[0030] Figure 10. shows the main interface of present invention present invention application in running mode

[0031] Figure 11 . Shows Records page of the present invention ’s application [0032] Figure 12. Shows Displaying records and their specific information

[0033] Figure 13. Shows Smartphone present invention ’s application pages

[0034] Figure 14. Illustrates the Homogeneous mixture diagnostic result page

[0035] Diagram 1 . Is detailed diagram explaining, The plot of light intensity for corresponding 1 to 7 McFarland standards.

A clear and precise statement of the advantages of the claimed invention over previous inventions

[0036] The presented device is able to measure the turbidity of the solution in the range of 0.5 to 7 McFarland units, which is in a high range of measurement accuracy of the Present invention unit. On the other hand, the presented device has the ability to detect instantaneously at high speed. This feature distinguishes the machine from existing models with low processing speed. Another important thing is the LED light source of the device. This light source replaces the UV light source used in similar devices with features such as biocompatibility, low energy consumption, stable light, easy switching, and cost-effectiveness. UV light source has disadvantages such as unreliable light, short life, high cost, and environmental pollution. Using a Raspberry Pi compatible camera greatly reduces the speed of response and measurement so that this operation is done online on this device.

Explanation of an executive method for applying an invention

[0037] First, to turn on the device, press the power button on the back of the device. After a few seconds, the device program is displayed on the LCD. Two locations are specified for the test sample and the control sample on the device. After placing the sample container in the specified position, the McFarland value is displayed by clicking on the run icon that appears on the LCD.

An explicit expression of the industrial application of the invention

[0038] The Present invention device is a system for detecting the turbidity of a solution and then counting the number of bacteria and cells in a solution. This system is used in medical diagnostic laboratories, microbiology, research centers, industrial centers, and food factories to determine the number of bacteria and cell in mixture solution. This device provides fast and accurate measurement of bacterial, cell, dispersion concentration for a routine and a large number of consecutive tests.]

Embodiments

[To better understand the current invention, the preferred embodiment is described as follows:

[0039] The present invention has been described the use of The accuracy of the density of McFarland Standards can be checked using a it with a 0.5 McFarland Standard has an absorbance reading with image processing system.

[0040] The present invention has been described turbidity of bacterial suspensions so that the number of bacteria will be within a given range to standardize microbial testing

[0041] The present invention has been described suspensions so that the number of cells will be within a given range to standardize cell number in suspension mixture determination

[0042] Another embodiment

[0043] is described for Al system is used for ratio on dispersion and homogeneity of hybrid liquid composite mixture.

[0044] Another embodiments is all the results, data and parameters that collected manually or automatically is stored in cloud based software that developed for device.

[0045] The present invention has been described with reference to particular embodiments having various features. In light of the disclosure provided above, it will be apparent to those skilled in the art that various modifications and variations can be made in the practice of present invention without departing from scope or spirit of invention. One skilled in the art will recognize that the disclosed features may be used singularly, in any combination, or omitted based on requirements and specifications of a given application or design. When an embodiment refers to “comprising” certain features, it is to be understood that the embodiments can alternatively ‘consist of ” or “consist essentially of’ any one or more of the features. Other embodiments of invention will be apparent to those skilled in the art from consideration of the specification and practice of the invention. [0046] Embodiments of the present invention will now be described in more detail with reference to the accompanying drawings. However, the present invention is not limited to the embodiments set forth herein and may be embodied in many different forms. Rather, these embodiments are provided so that this disclosure is thorough and complete, and will fully convey the scope of the invention to those skilled in the art. In the drawings, the sizes, such as widths and thicknesses, of elements may be exaggerated for clarity. The drawings are explained entirely from an observer's point of view. It will be understood that when an element is referred to as being “on” another element, it can be directly on the other element or one or more intervening elements may also be present there between. Those skilled in the art will appreciate that many modifications and variations can be made without departing from the spirit of the invention. Throughout the accompanying drawings, the same reference numerals are used to designate substantially the same elements.

[0047] On the other hand, terms used herein are to be understood as described below. The terms “first”, “second,” etc. are used only to distinguish one element from another and the scope of the claims should not be limited by these terms. For example, a first element could be termed a second element, and, similarly, a second element could be termed a first element.

[0048] It is noted in particular that where a range of values is provided in this specification, each value between the upper and lower limits of that range is also specifically disclosed. The upper and lower limits of these smaller ranges may independently be included or excluded in the range as well. The singular form “a”, ”an”, and “the” include plural referents unless the context clearly dictates otherwise. It is intended that the specification and examples be considers as exemplary in nature and that variations that do not depart from the essence of the invention fall within the scope of the invention. Further, all of the references cited in this disclosure are each individually incorporated by reference herein their entries and as such are intended to provide an efficient way of supplementing the enabling disclosure of this invention as well as provide background detailing level of ordinary skill the art.

[0049] Example 1 : The number of bacterial is diagnosed with device and the normal sample and the sample with Bactria is insert in chambers the specimen with intensity 218 and McFarland ratiol is calculated as the normal sample is diagnosed 0, another sample also McFarland 2 with intensity = 216 in another test as it shown in figure 8.

[0050] Example 2: when the device detect the McFarland ration there are two way to check the results, firstly the user can check the LCD and the result is recorded on it secondly through the Bluetooth connection with mobile application the main results are shown in The main interface of software application as it shown in figure 9. [0051] Example 3: The Displaying records and their specific information in software can be stored in cloud database and the client for example doctors, clinician, clinical staff or laboratorian can access and monitor all the tested samples as it shown in figure 11 , 12

[0052] Example 4: By adding one sample in detection chamber Homogeneous mixture diagnostic result is display in LCD with purity percentage index as it shown in figure 14 i

References

Claims

[Claim 1] I A hybrid diagnostic system method for this invented device comprising: a device comprising a two separated chamber for placement of each samples, a first sample source is homogenies sample without any additional components and the second chamber will cover second sample that contains specimen that should be tested ; a detection system configured for receipt of the device thereon or therein and for determining the amount of Bactria, cells as diagnostic instrument in communication with the Al detection system configured for analyzing desperation of components based on McFarland scale, a diagnostic instrument in communication with the detection system configured for analyzing a second sample with compared to the first sample source for one or more analysis.

[Claim 2] According to claim 1 ,the present invention uses science and technology such as image processing and Artificial intelligence , deep learning to rapidly and non- destructively measure solution turbidity and, by converting the turbidity obtained on the McFarland scale, it shows the number of bacteria and cells in the solution and determine the homogeneous solution of nanotube/solvent mixture, with no aggregation or precipitation.

[Claim 3] According to claim 1 , this device uses uniform LED light, which has properties such as stability, non-destructiveness, , to create a light beam source.

[Claim 4] According to claim 1 , the use of image processing technology has accelerated the measurement of turbidity and increased accuracy.

[Claim 5] This invented device comprising:

Firstly, the mechanical structure body that contains 2 holes for sample adjustments, mechanical holding for light adjustments, and isolated container for electrical and control board a device. The structure has two more containers that consist of five holes that the cover samples and can be stored before and after the diagnostic procedure.

Secondly, all electrical and electronically parts contain, LCD, LED Lights, wires, controllers, a camera, communication ports, modules such as WIFI, Bluetooth for communication with software application.

Finally, The software application that connect to device via Bluetooth or Wi-Fi and display all the results and store the results for telehealth control in database.

[Claim 6] According to claim 5, polylactic acid material is used in the construction of the device, which is biocompatible, portable and lightweight at the same time and the surface is coated with antibacterial composite.

[Claim 7] According to claim 5, the device structure is designed with ergonomically consideration and can be used as portable diagnostic device

[Claim 8] According to claim 5, The ability to separate tested and untested test tubes into two separate groups is another innovation of the mechanical part of this device.

[Claim 9] According to claim 5, with Al and Machin vision the present invention is analyzing and detect, Particle size distribution pattern and tensile properties of hybrid composite that correlated with the degree of dispersion.

[Claim 10] According to claim 5, this device has communication channels such as Bluetooth and WiFi to communicate with the mobile application for user interface.

[Claim 11] According to claim 9 , all the result data is stored in cloud server and is accessed by user application.]