Systems and methods for sample use maximization

• US 10,557,786 B2
• Filed: 05/19/2016
• Issued: 02/11/2020
• Est. Priority Date: 01/21/2011
• Status: Active Grant

• Current Assignee: Labrador Diagnostics LLC (SoftBank Group Corp.)
• Original Assignee: Theranos IP Company, LLC
• Inventors: Gibbons, Ian, Nugent, Anthony J., Delacruz, Anthony, Young, Daniel, Holmes, Elizabeth A., Drake, Andrew, Kemp, Timothy Michael, Balwani, Sunny, Pangarkar, Chinmay
• Primary Examiner(s): Ayub, Hina F
• Application Number:
  US15/159,501
• Publication Number:
  US 20160334403A1
• Time in Patent Office: 1,363 Days
• Field of Search: 356432
• CPC Class Codes: B01L 2300/168 Specific optical properties, e.g. reflective coatings
• CPC Class Codes: B01L 3/0275 Interchangeable or disposable dispensing tips
• CPC Class Codes: B04B 5/0421 pivotably mounted
• CPC Class Codes: C12Q 1/42 involving phosphatase
• CPC Class Codes: C12Q 1/48 involving transferase
• CPC Class Codes: C12Q 1/52 involving transaminase
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6809 Methods for determination or identification of nucleic acids involving differential detection
• CPC Class Codes: C12Q 1/689 for bacteria
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: G01N 1/4077 by other techniques involving separation of suspended solids
• CPC Class Codes: G01N 2001/4083 sedimentation
• CPC Class Codes: G01N 2035/00495 Centrifuges
• CPC Class Codes: G01N 2035/1018 Detecting inhomogeneities, e.g. foam, bubbles, clots
• CPC Class Codes: G01N 21/00 Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light G01N3/00 - G01N19/00 take precedence
• CPC Class Codes: G01N 21/0303 Optical path conditioning in cuvettes, e.g. windows; adapted optical elements or systems; path modifying or adjustment G01N21/031 - G01N21/15 take precedence
• CPC Class Codes: G01N 21/51 inside a container, e.g. in an ampoule G01N21/53 takes precedence
• CPC Class Codes: G01N 21/59 Transmissivity G01N21/25 takes precedence
• CPC Class Codes: G01N 2333/575 Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin G01N2333/665, corticotropin G01N2333/695
• CPC Class Codes: G01N 2333/62 Insulins
• CPC Class Codes: G01N 2333/91188 : transferring nitrogenous groups (2.6)
• CPC Class Codes: G01N 2333/96463 : Blood coagulation factors not provided for in a preceding group or according to more than one of the proceeding groups
• CPC Class Codes: G01N 33/5308 : for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
• CPC Class Codes: G01N 33/54326 : Magnetic particles
• CPC Class Codes: G01N 33/54393 : Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
• CPC Class Codes: G01N 33/56972 : White blood cells
• CPC Class Codes: G01N 33/56983 : Viruses
• CPC Class Codes: G01N 33/573 : for enzymes or isoenzymes
• CPC Class Codes: G01N 33/74 : involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
• CPC Class Codes: G01N 33/743 : Steroid hormones
• CPC Class Codes: G01N 33/80 : involving blood groups or blood types or red blood cells white blood cells G01N33/56972
• CPC Class Codes: G01N 33/82 : involving vitamins or their receptors
• CPC Class Codes: G01N 33/92 : involving lipids, e.g. cholesterol , lipoproteins, or their receptors steroid hormones G01N33/743
• CPC Class Codes: G01N 35/10 : Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices G01N35/0099 takes precedence
• CPC Class Codes: G06T 2207/30004 : Biomedical image processing
• CPC Class Codes: G06T 2207/30168 : Image quality inspection
• CPC Class Codes: G06T 3/4084 : in the transform domain, e.g. fast Fourier transform [FFT] domain scaling
• CPC Class Codes: G06T 7/0002 : Inspection of images, e.g. flaw detection
• CPC Class Codes: G06T 7/0012 : Biomedical image inspection
• CPC Class Codes: G16C 20/20 : Identification of molecular entities, parts thereof or of chemical compositions
• CPC Class Codes: H04N 23/64 : Computer-aided capture of images, e.g. transfer from script file into camera, check of taken image quality, advice or proposal for image composition or decision on when to take image

Abstract

The present invention provides systems, devices, and methods for point-of-care and/or distributed testing services. The methods and devices of the invention are directed toward automatic detection of analytes in a bodily fluid. The components of the device can be modified to allow for more flexible and robust use with the disclosed methods for a variety of medical, laboratory, and other applications. The systems, devices, and methods of the present invention can allow for effective use of samples by improved sample preparation and analysis.

Claims

1. A method of performing cytometry, comprising:

receiving in a sample processing device a cartridge comprising a blood sample obtained from a subject;

wherein the cartridge further comprises;

at least two pipette tips;

at least one assay unit;

an extraction tip;

a centrifugation vessel;

a cytometry cuvette, wherein the cytometry cuvette comprises at least two channels, wherein each channel comprises at least one sample entry port;

all reagents for performing cytometry, wherein the all reagents comprises at least two antibodies that bind to cell surface markers of white blood cells;

and wherein the sample processing device comprises;

a centrifuge;

a fluid transfer device comprising a pipette, wherein the pipette comprises a plurality of pipette heads, wherein a first pipette head and a second pipette head are configured to engage the cytometry cuvette and transport the cytometry cuvette from the cartridge to an imaging platform;

wherein the first pipette head and the second pipette head are configured to engage with a first pipette tip and a second pipette tip when not engaged with the cytometry cuvette;

the imaging platform configured to receive the cytometry cuvette, wherein the imaging platform comprises a light source, camera, and translation stage, wherein the translation stage is configured to move in the X, Y, and Z directions; and

an imaging device for imaging hematocrit of the blood sample;

engaging the first pipette head and the second pipette head of the pipette with the cytometry cuvette in the cartridge and transporting the cytometry cuvette from the cartridge to the imaging platform;

disengaging the first pipette head and the second pipette head from the cytometry cuvette;

engaging the first pipette head or the second pipette head of a liquid handling system with a pipette tip and transferring the blood sample to the centrifugation vessel;

centrifuging the blood sample in the centrifuge to obtain a plasma layer and a cell pellet;

measuring hematocrit of the centrifuged blood sample by imaging with the imaging device;

engaging the fluid transfer device with the at least one extraction tip and aspirating the plasma from the centrifuged blood sample;

resuspending the cell pellet and binding the at least two antibodies to the white blood cells in the cell pellet;

isolating the white blood cells and transferring with the fluid transfer device the white blood cells to the sample entry port of a channel of the cytometry cuvette; and

detecting the white blood cells with the camera of the imaging platform.

2. The method of claim 1, wherein the sample processing device further comprises a spectrometer.

3. The method of claim 1, wherein the imaging platform utilizes an image-based algorithm to control the z-position to achieve auto-focusing.

4. The method of claim 1, wherein the blood sample comprises about 200 μ

l or less.

5. The method of claim 1, wherein the blood sample is obtained by a fingerstick.

6. The method of claim 1, wherein the imaging platform provides epi-fluorescence illumination, darkfield illumination, and brightfield illumination.

7. The method of claim 6, wherein the imaging platform further comprises a filter wheel.

8. The method of claim 1, wherein each channel of the cytometry cuvette further comprises at least one air vent.

9. The method of claim 1, wherein each channel of the cytometry cuvette is about 10 to about 100 μ

m deep, about 0.5 to about 2 mm wide, and about 0.5 to about 5 cm long.

10. The method of claim 1, wherein the detection of the white blood cells comprises differentiating monocytes, lymphocytes, neutrophils, basophils, and eosinophils.

11. A blood sample processing system comprising:

a cartridge, wherein the cartridge comprises;

at least two pipette tips;

at least one assay unit;

an extraction tip;

a centrifugation vessel;

a cytometry cuvette, wherein the cytometry cuvette comprises at least two channels, wherein each channel comprises at least one sample entry port;

all reagents for performing cytometry, wherein the all reagents comprises at least two antibodies that bind to cell surface markers of white blood cells;

and a sample processing device, wherein the sample processing device comprises;

a fluid transfer device comprising a pipette, wherein the pipette comprises a plurality of pipette heads, wherein a first pipette head and a second pipette head are configured to engage the cytometry cuvette and transport the cytometry cuvette from the cartridge to an imaging platform, wherein the first pipette head and the second pipette head are configured to engage with a first pipette tip and a second pipette tip when not engaged with the cytometry cuvette, and wherein the first pipette head or the second pipette head is configured to transfer the blood sample to the centrifugation vessel after being engaged with the first pipette tip or the second pipette tip;

a centrifuge configured to centrifuge the blood sample to obtain a plasma layer and a cell pellet;

platform comprises a light source, camera, and translation stage, wherein the translation stage is configured to move in the X, Y, and Z directions; and

an imaging device configured to measure hematocrit of the blood sample using an imaging process, and configured to detect white blood cells isolated from the cell pellet using the camera.

12. The system of claim 11, wherein the device further comprises a spectrometer.

13. The system of claim 11, wherein each channel of the cytometry cuvette further comprises at least one air vent.

14. The system of claim 11, wherein each channel of the cytometry cuvette is about 10 to about 100 μ

m deep, about 0.5 to about 2 mm wide, and about 0.5 to about 5 cm long.

15. The system of claim 11, wherein the imaging platform provides epi-fluorescence illumination, darkfield illumination, and brightfield illumination.

16. The system of claim 11, wherein the imaging platform further comprises a filter wheel.

17. A blood sample processing device comprising:

a centrifuge configured to centrifuge the blood sample to obtain a plasma layer and a cell pellet;

a fluid transfer device comprising a pipette, wherein the pipette comprises a plurality of pipette heads, wherein a first pipette head and a second pipette head are configured to engage a cytometry cuvette and transport the cytometry cuvette from a cartridge to an imaging platform, wherein the first pipette head and the second pipette head are configured to engage with a first pipette tip and a second pipette tip when not engaged with the cytometry cuvette, and wherein at least one of the first pipette head or the second pipette head is configured to transfer the blood sample to the centrifuge for the centrifuging after being engaged with the first pipette tip or the second pipette tip;

the imaging platform configured to receive the cytometry cuvette, wherein the imaging platform comprises a light source, camera, and translation stage, wherein the translation stage is configured to move in the X, Y, and Z directions; and

an imaging device configured to measure hematocrit of the blood sample using an imaging process, and configured to detect white blood cells isolated from the cell pellet using the camera.

18. The device of claim 17, further comprising a spectrometer.

19. The device of claim 17, wherein the imaging platform provides epi-fluorescence illumination darkfield illumination and brightfield illumination.

20. The device of claim 17, wherein the imaging platform further comprises a filter wheel.