# US 10,769,773 B2 · Method and system for automated counting of microscopic fibers within a sample > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,769,773 B2 - Document ID: US10769773B2 - Status: Granted - Kind: B2 - Application number: 15/996,309 - Filed: 2018-06-01 - Published: 2019-05-02 - Granted: 2020-09-08 - Earliest claimed: 2017-06-02 - Assignees: Xrapid France - Applicants: Xrapid France - Inventors: David Mendels; Xavier Mulatou - Web page: https://patentgrep.com/patent/US10769773B2 ## CPC - G06T 7/0006: {using a design-rule based approach} - G01N 15/1468: {with spatial resolution of the texture or inner structure of the particle} - G02B 21/244: {using image analysis techniques} - G02B 21/26: Stages; Adjusting means therefor - G02B 21/362: {Mechanical details, e.g. mountings for the camera or image sensor, housings (G02B21/364 takes precedence)} - G02B 21/365: {Control or image processing arrangements for digital or video microscopes (G02B21/361, G02B21/362 take precedence)} - G06V 20/693: {Acquisition} - G06V 20/698: {Matching; Classification} - H04N 23/61: based on recognised objects - H04N 23/672: {based on the phase difference signals} - H04N 23/951: by using two or more images to influence resolution, frame rate or aspect ratio - G01N 2015/1486: {Counting the particles} - G02B 21/14: affording illumination for phase-contrast observation - G06T 2207/10056: Microscopic image - G06T 2207/10148: Varying focus - G06T 2207/30108: Industrial image inspection - G06T: IMAGE DATA PROCESSING OR GENERATION, IN GENERAL - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - G02B: OPTICAL ELEMENTS, SYSTEMS OR APPARATUS - G06V: IMAGE OR VIDEO RECOGNITION OR UNDERSTANDING - H04N: PICTORIAL COMMUNICATION, e.g. TELEVISION ## Abstract In an aspect of the disclosure, a method, a computer-readable medium, and an apparatus are provided. The method may include placing the sample on the sample-holder of a phase-contrast microscope, establishing a connection between the processor of the mobile terminal and a controller of the microscope, disposing a mobile terminal on a support so that its digital image sensor is positioned in front of an ocular, launching a counting program so that the processor performs successive counting sequences on distinct analysis areas of the sample, wherein between two counting sequences the processor controls a two-axis motorized system to displace the sample, and wherein for each counting sequence the processor implements the following steps of: setting the focus, capturing a main image, and analyzing the main image in order to count the number of fibers. ## Claims ### Claim 1 A counting method for automated counting of microscopic fibers within a sample, said counting method comprising the following steps of: providing a phase-contrast type microscope comprising an optical system provided with an ocular, an objective and a motorized focusing system, a sample-holder placed opposite the objective and displaceable according to two directions (X, Y) of an analysis plane by means of a two-axis motorized system, and a controller connected to the motorized focusing system and to the two-axis motorized system; providing a mobile terminal equipped with a digital image sensor, a screen, a processor and a memory in which is loaded a counting program; placing the sample on the sample-holder of the microscope; disposing the mobile terminal on a support of the microscope so that the digital image sensor is positioned and held stable in front of the ocular of the microscope with an alignment between the optical axis of the ocular and the center of the digital image sensor; establishing a connection between the processor of the mobile terminal and the controller of the microscope; launching the counting program so that the processor of the mobile terminal performs successive counting sequences on distinct analysis areas of the sample, wherein between two successive counting sequences the processor controls the two-axis motorized system to position the objective over a new analysis area of the sample, and wherein for each counting sequence the processor implements the following steps of: setting the focus on the corresponding analysis area comprising a control of the motorized focusing system; capturing a main image of the corresponding analysis area by triggering the digital image sensor; analyzing said main image in order to count the number of microscopic fibers on the corresponding analysis area. ### Claim 2 (depends on 1) The counting method according to claim 1, wherein, for each counting sequence, the setting of the focus implements an open-loop control in which the processor performs several successive focusing sequences, wherein between two successive focusing sequences the processor controls the motorized focusing system to modify the focus, and wherein for each focusing sequence the processor implements the following steps of: capturing a secondary image of the corresponding analysis area by triggering the digital image sensor; analyzing said secondary image in order to establish a value of a sharpness parameter representative of a sharpness of the secondary image; wherein the focus retained for the capture of the main image corresponds to the focus of the focusing sequence where the value of the sharpness parameter is maximum. ### Claim 3 (depends on 1) The counting method according to claim 1, wherein, for each counting sequence, the setting of the focus implements a closed-loop control in which the processor performs the following steps of: capturing a secondary image of the corresponding analysis area by triggering the digital image sensor; analyzing said secondary image in order to check whether a sharpness parameter, representative of a sharpness of the secondary image, reaches a predefined maximum threshold, where: if the sharpness parameter reaches or exceeds said maximum threshold, then the processor stops the control of the motorized focusing system in order to fix the focus for the capture of the main image; if the focus parameter is below said maximum threshold, then the processor controls the motorized focusing system to modify the focus and repeats the previous steps until the sharpness parameter reaches or exceeds said maximum threshold. ### Claim 4 (depends on 2) The counting method according to claim 2, wherein the processor performs the captures of the main images with a given main resolution, and performs the captures of the secondary images with a given secondary resolution which is lower than the main resolution. ### Claim 5 (depends on 2) The counting method according to claim 2, wherein the sharpness parameter corresponds to a ratio between a surface area of a halo region characteristic of a phase-contrast microscopy and a surface area of a central area surrounded by the halo region. ### Claim 6 (depends on 1) The counting method according to claim 1, wherein the processor stops the counting sequences when the number of microscopic fibers counted and cumulated throughout the counting sequences reaches a given fibers threshold or when the number of counting sequences reaches a given sequences threshold. ### Claim 7 (depends on 1) The counting method according to claim 1, wherein the processor displays on the screen of the mobile terminal the number of counted microscopic fibers and the number of performed counting sequences with an update as the counting sequences are carried out. ### Claim 8 (depends on 7) The counting method according to claim 7, wherein the number of counted microscopic fibers and the number of performed counting sequences are displayed in the form of two progress bars, wherein each progress bar is filled as the corresponding number increases until reaching the end of the progress bar which corresponds to the corresponding fibers threshold or sequences threshold, so that the processor stops the counting sequences as soon as one of the two progress bars is filled. ### Claim 9 (depends on 1) The counting method according to claim 1, wherein, for each counting sequence, the processor implements a multitask management distributed between a main queue and a background queue, where: the processor performs in the main queue the steps of capturing images and the steps of controlling the two-axis motorized system and the motorized focusing system, and the processor performs in the background queue the steps of analyzing the images, so that, between two consecutive counting sequences, the processor performs the control of the two-axis motorized system on the main queue before the end and in parallel with the analysis of the main image of the previous counting sequence which is performed on the background queue. ### Claim 10 (depends on 9) The counting method according to claim 9, wherein, between two successive counting sequences, the processor controls the two-axis motorized system according to a predefined scan path (CB) to position the objective over a new analysis area of the sample while care is taken that said new analysis area is distinct from each of the previously-analyzed analysis areas. ### Claim 11 (depends on 10) The counting method according to claim 10, wherein the predefined scan path (CB) is a random path. ### Claim 12 (depends on 1) The counting method according to claim 1, wherein the two-axis motorized system comprises: a first rotary motor driving in rotation a first output shaft on which is mounted a first pinion meshing with a first rack coupled to the sample-holder for a displacement according to a first direction (X); and a second rotary motor driving in rotation a second output shaft on which is mounted a second pinion meshing which a second rack coupled to the sample-holder for a displacement according to a second direction (Y); so that, when the processor controls the two-axis motorized system to displace the sample-holder, the processor controls at least one among the first rotary motor and the second rotary motor. ### Claim 13 (depends on 1) The counting method according to claim 1, wherein the two-axis motorized system ensures a stepped displacement of the sample-holder according to two directions so that, when the processor controls the two-axis motorized system to displace the sample-holder, the processor controls a stepped displacement of the sample-holder. ### Claim 14 (depends on 1) The counting method according to claim 1, wherein the counting method is implemented for automated counting of asbestos fibers within a sample. ### Claim 15 (depends on 1) The counting method according to claim 1, using as a mobile terminal a smartphone or a digital tablet or a personal digital assistant. ### Claim 16 (depends on 1) The counting method according to claim 1, wherein the connection between the processor of the mobile terminal and the controller of the microscope is established: either in a wired manner by means of a wired connector; or in a wireless manner by means of a radiocommunication in particular a short-range radiocommunication such as a for example a Bluetooth™, Bluetooth Low Energy™ or Zigbee™ radiocommunication. ### Claim 17 A microscope adapted for the implementation of the counting method according to any one of the preceding claims, said microscope being a phase-contrast microscope comprising: an optical system provided with an ocular, an objective and a motorized focusing system, a sample-holder placed opposite the objective and displaceable according to two directions by means of a two-axis motorized system, a controller connected to the motorized focusing system and to the two-axis motorized system; and a support mounted on the optical system at the level of the ocular, said support being shaped so as to support a mobile terminal equipped with a digital image sensor so that said digital image sensor is positioned and held stable in front of the ocular of the microscope with an alignment between the optical axis of the ocular and the center of the digital image sensor. ### Claim 18 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16) A system for automated counting of microscopic fibers within a sample, said automated counting system comprising: a microscope adapted for the implementation of the counting method according to any one of the preceding claims, said microscope being a phase-contrast microscope including: an optical system provided with an ocular, an objective and a motorized focusing system, a sample-holder placed opposite the objective and displaceable according to two directions by means of a two-axis motorized system, a controller connected to the motorized focusing system and to the two-axis motorized system; and a support mounted on the optical system at the level of the ocular, said support being shaped so as to support a mobile terminal equipped with a digital image sensor so that said digital image sensor is positioned and held stable in front of the ocular of the microscope with an alignment between the optical axis of the ocular and the center of the digital image sensor; a mobile terminal equipped with a digital image sensor, a screen, a processor and a memory in which is loaded a counting program configured to perform the steps implemented by the processor in the counting method according to any one of claims 1 to 16; wherein the support of the microscope supports said mobile terminal so that said digital image sensor is positioned and held stable in front of the ocular of the microscope with an alignment between the optical axis of the ocular and the center of the digital image sensor. ### Claim 19 (depends on 1) A non-transitory computer-readable medium adapted to be loaded in a memory of a mobile terminal and comprising code instructions capable of controlling the execution of the steps implemented by the processor in the counting method according to claim 1 when it is executed on the mobile terminal. ### Claim 20 (depends on 19) The non-transitory computer-readable medium of claim 19, wherein the computer-readable medium is downloadable on a telecommunication network in a memory of a mobile terminal.