Method and device for detecting bacteria and determining the concentration thereof in a liquid sample

• US 10,359,352 B2
• Filed: 03/16/2017
• Issued: 07/23/2019
• Est. Priority Date: 10/29/2013
• Status: Active Grant

• Current Assignee: IDEXX Laboratories Incorporated
• Original Assignee: IDEXX Laboratories Incorporated
• Inventors: Hammond, Jeremy, Russell, James
• Primary Examiner(s): Gurtowski, Richard C
• Application Number:
  US15/461,087
• Publication Number:
  US 20170212032A1
• Time in Patent Office: 859 Days
• Field of Search: None
• CPC Class Codes: B01L 2200/0652 Sorting or classification of particles or molecules
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0851 Bottom walls
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2400/086 using baffles or other fixed flow obstructions
• CPC Class Codes: B01L 3/502 with fluid transport, e.g. in multi-compartment structures centrifugal-type cuvettes G01N21/07; analysis by separation into components G01N30/00; automatic analysers G01N35/00
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: C12Q 1/06 Quantitative determination
• CPC Class Codes: G01N 15/01 specially adapted for biological cells, e.g. blood cells investigating sedimentation of particle suspensions in blood G01N15/05
• CPC Class Codes: G01N 15/14 Optical investigation techniques, e.g. flow cytometry
• CPC Class Codes: G01N 15/149 specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: G01N 2015/1006 for cytology
• CPC Class Codes: G01N 2015/1447 Spatial selection
• CPC Class Codes: G01N 2015/1452 Adjustment of focus; Alignment
• CPC Class Codes: G01N 2015/1486 Counting the particles
• CPC Class Codes: G01N 2333/195 from bacteria
• CPC Class Codes: G01N 33/493 urine

Abstract

A method for detecting bacteria and determining the concentration thereof in a liquid sample includes the steps of taking an optical section through a container holding a volume of the liquid sample at a predetermined field of view and at a predetermined focal plane depth or angle and after a period of time has elapsed to allow non-bacteria in the sample to settle to the bottom of the container. Since bacteria auto arranges in the liquid sample, forming a lattice-like grid pattern, an optical section through the volume of auto-arranged bacteria may be used to measure the quantity of bacteria residing in that section. A container for holding the liquid sample has particular structure which aids in separating the non-bacteria from the bacteria.

Claims

1. A container for holding a volume of a liquid sample and used for separating different types of particles within the volume of the liquid sample, the different types of particles within the volume of the liquid sample held by the container including a first type of particles which auto arrange within the volume of the liquid sample held by the container and a second type of particles which do not auto arrange within the volume of the liquid sample, the container comprising:

a bottom wall, the bottom wall having a recessed portion and a non-recessed portion adjacent the recessed portion, the container thereby defining a first zone situated at a first depth in the volume of the liquid sample and in vertical alignment with the non-recessed portion of the container bottom wall, and a second zone situated at a second depth in the volume of the liquid sample and in vertical alignment with the recessed portion of the container bottom wall, the first type of particles which auto arrange substantially occupying the second zone within the container, and the second type of particles which do not auto arrange substantially occupying the first zone within the container; and

at least one projection, the at least one projection extending upwardly from the non-recessed portion of the container bottom wall and at least partially into the volume of the liquid sample held by the container, the at least one projection being situated on the non-recessed portion of the bottom wall in proximity to the recessed portion of the bottom wall, the at least one projection further acting to separate the first type of particles which auto arrange and which substantially occupy the second zone within the container from the second type of particles which do not auto arrange and substantially occupy the first zone within the container.

2. A container for holding a volume of a liquid sample and used for separating bacteria which auto arrange within the volume of the liquid sample held by the container and non-bacteria which do not auto arrange within the volume of the liquid sample, the container comprising:

a container bottom surface;

a container top surface, the container bottom surface and the container top surface together defining a fluid flow channel through which the liquid sample held by the container may flow in an axial direction through the container; and

a plurality of projections spaced apart from each other over at least a portion of the axial length of the container, the projections extending upwardly from the bottom surface of the container and at least partially into the fluid flow channel and the volume of the liquid sample held thereby, the projections being arranged such that one or more of the projections are in mutually axial alignment within the container, whereby at least one projection is situated in axial alignment and spaced apart from a respective adjacent projection successively in the direction of flow of the liquid sample in the fluid flow channel, the projections defining a first zone and at least a second zone adjacent the first zone, bacteria which auto arrange substantially occupying the first zone within the container, and the non-bacteria which do not auto arrange substantially occupying the at least second zone within the container.

6. A container as defined by claim 2, wherein all of the projections are in mutually axial alignment within the container.

3. A container for holding a volume of a liquid sample and used for detecting bacteria which auto arrange within the volume of the liquid sample held by the container or have a first dimension, and non-bacteria which do not auto arrange within the volume of the liquid sample held by the container or have a second dimension which is different from the first dimension of the bacteria, the container comprising:

a bottom wall; and

an optical focus reference structure including at least one of an arrangement of a plurality of spaced apart recesses, an arrangement of a plurality of spaced apart projections or a fixed bead or latex particle or combinations thereof formed in the bottom wall or situated in proximity to the bottom wall of the container.

4. A container for holding a volume of a liquid sample and used for detecting bacteria which auto arrange within the volume of the liquid sample held by the container or have a first dimension, and non-bacteria which do not auto arrange within the volume of the liquid sample held by the container or have a second dimension which is different from the first dimension of the bacteria, the container comprising:

a bottom wall;

a first lateral side wall;

a second lateral side wall situated opposite to and in parallel with the first lateral side wall;

a first axial end;

a second axial end situated opposite the first axial end; and

a plurality of parallelly disposed and spaced apart plates, the plates being joined to and extending perpendicularly upwardly from the bottom wall of the container and at least partially into the volume of the liquid sample held by the container, the plurality of plates including adjacent plates situated in a side-by-side, parallel, spaced apart arrangement between the first lateral side wall and the second lateral side wall of the container, each plate having a lateral side which faces the lateral side of a respective adjacent plate, the adjacent plates of the plurality of parallelly disposed plates defining fluid flow channels therebetween to direct the flow of liquid sample between adjacent plates in a direction between the first axial end and the second axial end of the container.

5. A container for holding a volume of a liquid sample as defined by claim 4,

wherein at least some of the plates of the plurality of parallelly disposed plates have differing axial lengths which increase from the longitudinal center of the container in symmetrical directions outwardly toward the opposite first lateral side and second lateral side thereof.