Particle identification system, cartridge and associated methods

• US 9,739,714 B2
• Filed: 03/15/2013
• Issued: 08/22/2017
• Est. Priority Date: 10/29/2012
• Status: Active Grant

• Current Assignee: MBio Diagnostics, Inc. (Mars Incorporated)
• Original Assignee: MBio Diagnostics, Inc. (Mars Incorporated)
• Inventors: Moll, Kevin D., Delaney, Marie J., Vogel, Kurt R., Lochhead, Michael J., Ives, Jeffrey, Dunn, John S., Rowley, Keagan B.
• Primary Examiner(s): Crawford, Erik B
• Application Number:
  US13/831,757
• Publication Number:
  US 20140120556A1
• Time in Patent Office: 1,621 Days
• CPC Class Codes: B01L 2200/0684 Venting, avoiding backpressure, avoid gas bubbles
• CPC Class Codes: B01L 2200/12 Specific details about manufacturing devices
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/044 pierceable, e.g. films, membranes
• CPC Class Codes: B01L 2300/045 whereby the whole cover is slidable
• CPC Class Codes: B01L 2300/046 Function or devices integrated in the closure
• CPC Class Codes: B01L 2300/048 enabling gas exchange, e.g. vents
• CPC Class Codes: B01L 2300/0627 Sensor or part of a sensor is integrated
• CPC Class Codes: B01L 2300/0672 Integrated piercing tool
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0838 Capillaries
• CPC Class Codes: B01L 2300/161 Control and use of surface tension forces, e.g. hydrophobic, hydrophilic
• CPC Class Codes: B01L 2300/168 Specific optical properties, e.g. reflective coatings
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0694 vents used to stop and induce flow, backpressure valves
• CPC Class Codes: B01L 2400/086 using baffles or other fixed flow obstructions
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/502723 characterised by venting arrangements
• CPC Class Codes: B01L 3/502746 characterised by the means for controlling flow resistance, e.g. flow controllers, baffles B01L3/502738 takes precedence
• 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: G01N 21/6428 : Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6456 : Spatial resolved fluorescence measurements; Imaging
• CPC Class Codes: G01N 33/54366 : Apparatus specially adapted for solid-phase testing

Abstract

A particle identification system includes: a cartridge for containing a sample with fluorescently labeled particles; illumination for illuminating a region within the cartridge to stimulate emission from particles; imager for generating wavelength-filtered electronic images of the emission within at least one measurement field of the region; and particle identifier for processing the electronic images to determine a superset of particles of interest, and fluorescently labeled particles within the superset based on properties of the particles in the at least one measurement field. A method determines fluorescently labeled particles within a sample, by: processing at least one electronic image from at least one focal position within the sample; determining dimmest separation lines between brighter areas in the electronic image; and, for each of the brighter areas, determining local background level based on pixel values of the separation lines forming a perimeter therearound, to determine each of the fluorescently labeled particles.

Claims

1. Cartridge for detecting target analytes in a sample, comprising:

a capillary channel with a detection region for detecting the target analytes;

an inlet port for holding the sample before entering the capillary channel and including dried reagents for rehydrating into the sample, the inlet port and the capillary channel being shaped in part by a common structure;

a vent having a frangible seal for preventing flow of the sample into the capillary channel until the frangible seal is broken; and

a movable lid positionable in an open position, to provide access to the inlet port, and a closed position, to prevent access to the inlet port, the lid including a protrusion that punctures the frangible seal when the lid is moved from the open position to the closed position.

2. Cartridge of claim 1, the dried reagents having a rehydration rate determined by reagent formulation and physical dimension that collectively yield spatially uniform reagent-sample interactions within the detection region.

3. Cartridge of claim 1, the dried reagents being a dried reagent coating deposited as a pattern of liquid reagent formulation droplets that merge together and dry to form the dried reagent coating.

4. Cartridge of claim 3, each of the liquid reagent formulation droplets consisting of nanoliters in volume, the liquid reagent formulation droplets deposited with center-to-center spacing that ensures, together with the volume, droplet-to-droplet contact following deposition and prior to drying to form the dried reagent coating.

5. Cartridge of claim 4, the center-to-center spacing being 0.5 millimeters, the pattern being an evenly distributed pattern.

6. Cartridge of claim 1, the capillary channel having a volume capacity of microliters, the inlet port having a volume capacity greater than the volume capacity of the capillary channel.

7. Cartridge of claim 1, the capillary channel being formed in part by one optically transmissive planar substrate and a planar inner surface of a cartridge body.

8. Cartridge of claim 1, further comprising control features disposed to be viewable within the detection region, to permit evaluation of assay validity.

9. Cartridge of claim 1, further comprising a machine-readable label encoded with height of the capillary channel and viewable by instrumentation to calibrate use of the cartridge according to the height, the height being an internal dimension of the capillary channel.

10. Cartridge of claim 1, further comprising a locking mechanism for irreversibly closing the lid in the closed position.

11. Cartridge of claim 1, the capillary channel being at least in part hydrophilic to facilitate movement of the sample therethrough.

12. The cartridge of claim 1, further comprising a planar substrate for forming the bottom surface and a floor of the capillary channel, the capillary channel having height orthogonal to the floor and the flow direction, the height being sized to restrict flow, of the sample within the capillary channel, to capillary flow.

13. The cartridge of claim 12, the inlet port having volume capacity greater than volume of the sample required to detect the target analytes.

14. The cartridge of claim 13, the inlet port having hydrophilic surfaces that generate capillary forces to acquire the sample when the inlet port, in a downwards facing orientation, touches the sample.

15. The cartridge of claim 1, wherein an entrance from the inlet port into the capillary channel is shaped to seal the sample at the entrance and prevent flow of the sample into the capillary channel when the sample is located in the inlet port and the frangible seal is not broken.

16. The cartridge of claim 12, the height of the capillary channel being constant along length of capillary channel from the inlet port through the detection region to the vent.

17. The cartridge of claim 1, the inlet port and the vent facing in same direction.