Instrument and system for rapid microorganism identification and antimicrobial agent susceptibility testing

• US 10,273,521 B2
• Filed: 12/20/2017
• Issued: 04/30/2019
• Est. Priority Date: 03/30/2015
• Status: Active Grant

• Current Assignee: Accelerate Diagnostics, Inc.
• Original Assignee: Accelerate Diagnostics, Inc.
• Inventors: Ashby, Austin, Showalter, Wayne, Cote, Jasmin
• Primary Examiner(s): Bhat, Narayan K
• Application Number:
  US15/849,297
• Publication Number:
  US 20180135093A1
• Time in Patent Office: 496 Days
• CPC Class Codes: B01L 2300/0893 having a very large number of wells, microfabricated wells
• CPC Class Codes: B01L 3/527 for a plurality of reagents
• CPC Class Codes: C12Q 1/025 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics antimicrobial activity C12Q1/18
• CPC Class Codes: C12Q 1/04 Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor C12Q1/6897 takes precedence
• CPC Class Codes: G01N 1/38 Diluting, dispersing or mixing samples
• CPC Class Codes: G01N 15/06 Investigating concentration of particle suspensions by weighing G01N5/00; investigating sedimentation of particle suspensions G01N15/04; investigating individual particles G01N15/10
• CPC Class Codes: G01N 15/075 by optical means
• CPC Class Codes: G01N 2021/6419 Excitation at two or more wavelengths
• CPC Class Codes: G01N 2035/00346 Heating or cooling arrangements
• CPC Class Codes: G01N 2035/0443 for reagents
• CPC Class Codes: G01N 2035/0444 for cuvettes or reaction vessels
• CPC Class Codes: G01N 2035/0446 Combinations of the above
• CPC Class Codes: G01N 2035/0455 Coaxial carousels
• CPC Class Codes: G01N 2035/1032 Dilution or aliquotting
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 35/025 having a carousel or turntable for reaction cells or cuvettes
• CPC Class Codes: G01N 35/1002 Reagent dispensers
• CPC Class Codes: G02B 7/28 Systems for automatic generation of focusing signals

Abstract

A system for automated microorganism identification and antibiotic susceptibility testing comprising a reagent cartridge, a reagent stage, a cassette, a cassette, stage, a pipettor assembly, an optical detection system, and a controller is disclosed. The system is designed to dynamically adjust motor idle torque to control heat load and employs a fast focus process for determining the true focus position of an individual microorganism. The system also may quantify the relative abundance of viable microorganisms in a sample using dynamic dilution, and facilitate growth of microorganisms in customized media for rapid, accurate antimicrobial susceptibility testing.

Claims

1. A system for autofocusing on a microorganism to be imaged, comprising:

a first light source for projecting a beam of light and generating a reflective spot on an image plane;

a second light source for projecting light to capture an image of the microorganism on the image plane;

a movable objective positioned between the first and second light sources and the image plane for allowing focusing on the image plane, the movable objective being movable to a first out-of-focus position having a first focal point a first predetermined distance in front of the image plane and to a second out-of-focus position having a second focal point a second predetermined distance behind the image plane;

an image sensor for capturing the reflective spot and for capturing the image of the microorganism using the objective to focus; and

a controller for receiving two or more images of the reflective spot from the image sensor, wherein the two or more images are captured with the objective in at least the first and second out-of-focus positions, and for modifying a position of the objective based on intensity weightings of the reflective spots to an in-focus position.

2. The system of claim 1, wherein the intensity weightings relate to a greyscale value associated with each pixel that is weighted according to each pixel'"'"'s position relative to a center.

3. The system of claim 1, wherein the controller is configured to perform the following:

perform a prescan wherein intensity weightings over a plurality of focus positions of the objective are acquired.

4. The system of claim 1, wherein the controller is configured to perform the following:

perform a prescan wherein offset versus focus position is obtained for a plurality of focus positions of the objective.

5. The system of claim 1, wherein the controller is configured to perform the following:

calculate a focus position using the intensity weightings of the reflective spots together with the prescan of the intensity weightings to determine the position of the objective.