# US 10,900,877 B1 · Methods, apparatus, and systems to extract and quantify minute objects from soil or feces, including plant-parasitic nematode pests and their eggs in soil > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,900,877 B1 - Document ID: US10900877B1 - Status: Granted - Kind: B1 - Application number: 15/914,735 - Filed: 2018-03-07 - Granted: 2021-01-26 - Earliest claimed: 2017-03-08 - Assignees: Iowa State University Research Foundation, Inc. - Applicants: Iowa State University Research Foundation, Inc. - Inventors: Santosh Pandey; Augustine Beeman; Leland E. Harker; Jared P. Jensen; Upender Kalwa; Taejoon Kong; Zach L. Njus; Gregory L. Tylka; Christopher M. Legner - Web page: https://patentgrep.com/patent/US10900877B1 ## CPC - G01N 1/286: {involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising (microtomes G01N1/06; pulverising in general B02C; mixing in general B01F)} - G01N 1/34: Purifying; Cleaning {(processes or apparatus for extracting or separating nucleic acids from biological samples C12N15/1003)} - G01N 1/38: Diluting, dispersing or mixing samples - G01N 1/40: Concentrating samples - G01N 35/00871: {Communications between instruments or with remote terminals} - G01N 35/0099: {comprising robots or similar manipulators (robots per se B25J)} - G01N 35/02: using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations {(G01N35/0098 and G01N35/0099 take precedence)} - G01N 35/04: Details of the conveyor system {(G01N35/021 - G01N35/028 take precedence)} - G05B 19/402: characterised by control arrangements for positioning, e.g. centring a tool relative to a hole in the workpiece, additional detection means to correct position (G05B19/19 takes precedence) - G06T 7/0012: {Biomedical image inspection} - G06V 10/26: Segmentation of patterns in the image field; Cutting or merging of image elements to establish the pattern region, e.g. clustering-based techniques; Detection of occlusion - G06V 10/50: by performing operations within image blocks; by using histograms, e.g. histogram of oriented gradients [HoG]; by summing image-intensity values; Projection analysis - G06V 10/751: {Comparing pixel values or logical combinations thereof, or feature values having positional relevance, e.g. template matching} - G06V 20/69: Microscopic objects, e.g. biological cells or cellular parts - G06V 20/693: {Acquisition} - G01N 2001/2866: {Grinding or homogeneising} - G01N 2035/0441: {for samples} - G05B 2219/23416: Enter application program into I-O module, like motion program, servo program - G06T 2207/10016: Video; Image sequence - G06T 2207/10056: Microscopic image - G06T 2207/30188: Vegetation; Agriculture - G06T 2207/30242: Counting objects in image - 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) - G05B: CONTROL OR REGULATING SYSTEMS IN GENERAL; FUNCTIONAL ELEMENTS OF SUCH SYSTEMS; MONITORING OR TESTING ARRANGEMENTS FOR SUCH SYSTEMS OR ELEMENTS (systems for controlling or regulating non-electric variables G05D; systems for regulating electric or magnetic variables G05F; control devices or systems insofar as characterised by mechanical features only G05G) - G06T: IMAGE DATA PROCESSING OR GENERATION, IN GENERAL - G06V: IMAGE OR VIDEO RECOGNITION OR UNDERSTANDING ## Abstract A system, method, and apparatus for quantification of pre-determined particles in a soil or feces sample with certain automated steps. In one aspect it includes an input station for inputting a soil or feces sample; a sieving/filtering station for separating soil or feces from particles or particle carriers, and/or separating particle carriers from particles; and a collection station for receiving the extracted particles. It can include quantification of the collected sample with an imaging station to digitally image the particles and recognize and count the particles collected. A mechanism can mechanically move the filtered particles from the sieve/filter station to the collection station. A controller can be programmed to automatically control at least certain functions of the mechanism. An optional feature includes acquisition of chemical, biological, physical, or other parameters of the sample with one or more sensors positioned or positionable at the sample. ## Claims ### Claim 1 A system for extraction and quantification of pre-determined objects of interest in a soil or feces sample with automated steps comprising: a. an extraction station comprising: i. a motion-controlled rotatable multilevel turntable assembly with rotational position feedback, wherein each level of the multilevel turntable assembly comprises at least one receiver for removable insertion of a sieve; ii. a motion-controlled sieve manipulator to position sieves of different screen sizes relative to one another for at least one sieving action on the soil or feces sample to separate and extract the objects of interest from the remainder of the soil or feces sample, wherein the motion-controlled sieve manipulator has a range of movement to each level of the multilevel turntable adapted for controlled movement of sieves in position relative to other components of the system or relative to space generally; and iii. a collection container for receiving the extracted objects of interest; b. a quantification station for quantifying the extracted objects of interest comprising: i. an imaging location adapted to present the extracted objects of interest for imaging; ii. an image recognition or reconstruction sub-system having a field of view including the imaging location and pattern recognition software programmed with pattern-recognition parameters correlated to the objects of interest; and c. a programmable controller for instructing motion control in the extraction station; d. so that one or more of the following can be conducted on a soil or feces sample: i. objects of interest are automatically extracted from the soil or feces sample; and ii. extracted objects of interest are automatically counted by pattern recognition. ### Claim 2 (depends on 1) The system of claim 1 wherein the extraction station further comprises: a. at least two vertically positioned sieves of different pore sizes that are supported on the multilevel turntable. ### Claim 3 (depends on 1) The system of claim 1 further comprising a grinder tool at the extraction station having a rotatable grinding pad sized to fit in a sieve and adapted to provide gentle grinding and press actions on a sample on a sieve screen. ### Claim 4 (depends on 1) The system of claim 1 further comprising a washing tool adapted to provide washing liquid to a sieve, wherein the washing tool is motion-controlled for movement in the extraction station. ### Claim 5 (depends on 1) The system of claim 1 further comprising an input station, the input station comprising: a. a conveyor sub-system to support and automatically transport a plurality of soil or feces samples to the extraction station; b. a collection component to receive extracted particles and contain them in a container that can be identified and correlated to the input sample. ### Claim 6 (depends on 1) The system of claim 1 wherein objects of interest comprise: a. nematodes, wherein the different screen sizes include at least one to pass nematode cysts and one to pass nematode eggs; or b. one of fungal spores, weed seeds, eggs of nematodes, larvae of nematodes, cysts of nematodes, organisms, eggs of insects, larvae of insects, and soil mites. ### Claim 7 (depends on 1) The system of claim 1 wherein the image recognition subsystem comprises: a. a digital imager for static imaging; b. a substrate to support extracted objects of interest in the digital scanner; c. software to; i. scan discrete areas of the substrate, ii. store images of each discrete area, iii. run pattern recognition on each discrete area; iv. increment a counter for each pattern recognition indicative of an object of interest; and v. provide a final cumulative count. ### Claim 8 (depends on 1) The system of claim 1 wherein the image recognition subsystem comprises: a. a lens-free photodetector array sensor for video imaging; b. a microfluidic subsystem to flow extracted objects of interest in solution past the imaging location of the lens-free photodetector array sensor to acquire video of the flow c. software to; i. select two frames of video; ii. run image reconstruction on each of the frames; iii. run pattern recognition on each reconstruction; iv. increment a counter for each pattern recognition indicative of an object of interest object of interest, and v. provide a final cumulative count. ### Claim 9 (depends on 1) The system of claim 1 further comprising: a. a sensor subassembly including one or more sensors adapted to sense one or more attributes of the sample and communicate a sensed attribute to a processor; b. the sensor subassembly being embodied in at least one of: i. a sieve holding a sample; or ii. a motion-controlled tool that can be moved to a sieve holding a sample. ### Claim 10 (depends on 1) The system of claim 1 wherein the sample is processed prior to imaging by density separation. ### Claim 11 A method to automate extraction of objects of interest from a soil or feces sample comprising: a. preparing a diluted soil or feces sample containing objects of interest; b. conducting a first sieving on a first sieve for separating the objects of interest from larger particles in the soil or feces sample with automated washing action, wherein the first sieving comprises controlled movement of the first sieve in position relative to other components of the system or relative to space generally; c. collecting the separated objects of interest on a second sieve; d. conducting a second sieving on the second sieve to influence releasing the objects of interest and passing them through the second sieve, wherein the second sieving comprises controlled movement of the second sieve both in position relative to other components of the system or relative to space generally; and e. collecting what is passed through the second sieve. ### Claim 12 (depends on 11) The method of claim 11 further comprising conducting one or more of: a. a third sieving with a third sieve after the second sieve, the third sieve having a screen size to pass objects of interest; and b. spraying water in and on the second sieve between periods of grinding and pressing. ### Claim 13 (depends on 11) The method of claim 11 further comprising quantifying the collected objects of interest by automated digital pattern recognition of acquired images of the objects of interest. ### Claim 14 (depends on 13) The method of claim 13 wherein the acquired images are scans of a static substrate containing the objects of interest, wherein recognized patterns indicative of an object of interest are counted for the acquired images. ### Claim 15 (depends on 13) The method of claim 13 wherein the acquired images are video frames of flowing solution containing the objects of interest, wherein recognized patterns indicative of an object of interest are counted for the acquired images. ### Claim 16 (depends on 15) The method of claim 15 wherein the count is of objects of interest comprising one of nematode worms, cysts, or eggs and used to estimate an infestation level for the soil or feces sample, further comprising using the estimate for remedial action at the source of the soil or feces sample. ### Claim 17 (depends on 11) The method of claim 11 further comprising automatically sensing a chemical, physical, electrical, electromagnetic, optical, radiometric, mechanical, acoustic, pneumatic, electrochemical or biological parameter of the sample or the objects of interest in the sample by use of one or more sensors. ### Claim 18 (depends on 11) The method of claim 11 wherein the sample is processed prior to imaging by density separation. ### Claim 19 A system for extraction and quantification of pre-determined objects of interest in a soil or feces sample with automated steps comprising: a. an extraction station comprising: i. a motion-controlled sieve manipulator to position sieves of different screen sizes relative to one another for at least one sieving action on the soil or feces sample to separate and extract the objects of interest from the remainder of the soil or feces sample; ii. a collection container for receiving the extracted objects of interest; b. a quantification station for quantifying the extracted objects of interest comprising: i. an imaging location adapted to present the extracted objects of interest for imaging; ii. an image recognition or reconstruction sub-system having a field of view including the imaging location and pattern recognition software programmed with pattern-recognition parameters correlated to the objects of interest; wherein the image recognition or reconstruction subsystem comprises: a. a lens-free photodetector array sensor for video imaging; b. a microfluidic subsystem to flow extracted objects of interest in solution past the imaging location of the lens-free photodetector array sensor to acquire video of the flow c. software to; i. select two frames of video; ii. run image reconstruction on each of the frames; iii. run pattern recognition on each reconstruction; iv. increment a counter for each pattern recognition indicative of an object-of interest, and v. provide a final cumulative count; and c. a programmable controller for instructing motion control in the extraction station; d. so that one or more of the following can be conducted on a soil or feces sample: i. objects of interest are automatically extracted from the soil or feces sample; and ii. extracted objects of interest are automatically counted by pattern recognition. ### Claim 20 (depends on 19) The system of claim 19 wherein the extraction station further comprises: a. a motion-controlled rotatable multilevel turntable with rotational position feedback; b. each level comprising at least one receiver for removable insertion of a sieve; and c. at least two vertically positioned sieves of different pore sizes are supported on the multilevel turntable. ### Claim 21 (depends on 19) The system of claim 19 wherein the motion-controlled sieve manipulator has range of movement to each level of the multilevel turntable. ### Claim 22 (depends on 19) The system of claim 19 further comprising a grinder tool at the extraction station having a rotatable grinding pad sized to fit in a sieve and adapted to provide gentle grinding and press actions on a sample on a sieve screen. ### Claim 23 (depends on 19) The system of claim 19 further comprising a washing tool adapted to provide washing liquid to a sieve, wherein the washing tool is motion-controlled for movement in the extraction station. ### Claim 24 (depends on 19) The system of claim 19 further comprising an input station, the input station comprising: a. a conveyor sub-system to support and automatically transport a plurality of soil or feces samples to the extraction station; b. a collection component to receive extracted particles and contain them in a container that can be identified and correlated to the input sample. ### Claim 25 (depends on 19) The system of claim 19 wherein objects of interest comprise: a. nematodes, wherein the different screen sizes include at least one to pass nematode cysts and one to pass nematode eggs; or b. one of fungal spores, weed seeds, eggs of nematodes, larvae of nematodes, cysts of nematodes, organisms, eggs of insects, larvae of insects, and soil mites.