Nuclear morphology based identification and quantification of white blood cell types using optical bio-disc systems
First Claim
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1. A method of conducting an assay employing an optical disc and disc drive, said method comprising the steps of:
- providing a sample of cells on a disc surface in a chamber in a disc, the chamber including at least one capture zone with a capture agent;
loading the disc into an optical reader;
rotating the optical disc;
directing an incident beam of electromagnetic radiation to the capture zone;
detecting a beam of electromagnetic radiation formed after interacting with the disc at the capture zone;
converting the detected beam into an output signal; and
analyzing the output signal to extract therefrom information relating to the number and type of cells captured at the capture zone.
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Abstract
The present invention relates in general to biological assays and diagnostic assays and, in particular, to methods and apparatuses for imaging cells using an optical bio-disc system. More specifically, but without restriction to the particular embodiments hereinafter described in accordance with the best mode of practice, this invention relates to methods for identifying and quantitating cells including white blood cells based on the morphology of their nucleus using stains that absorb electromagnetic radiation at pre-determined wavelengths in conjunction with optical bio discs.
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Citations
40 Claims
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1. A method of conducting an assay employing an optical disc and disc drive, said method comprising the steps of:
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providing a sample of cells on a disc surface in a chamber in a disc, the chamber including at least one capture zone with a capture agent;
loading the disc into an optical reader;
rotating the optical disc;
directing an incident beam of electromagnetic radiation to the capture zone;
detecting a beam of electromagnetic radiation formed after interacting with the disc at the capture zone;
converting the detected beam into an output signal; and
analyzing the output signal to extract therefrom information relating to the number and type of cells captured at the capture zone.
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2. An optical disc and drive system for receiving a sample, the system comprising:
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a disc including a substrate, a cap parallel to the substrate, and a chamber defined between said cap and substrate including capture zones;
a capture layer over the substrate at the capture zones, a first capture zone having first cell capture agents and a second capture zone having a second cell capture agents;
a light source for directing light toward the disc at the capture zones;
a detector for detecting light reflected from or transmitted through the disc at the capture zones and providing a signal; and
a processor for using the signal to distinguish the nuclei of captured cells and count items in the sample bound to the capture molecules.
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3. A method of performing a white blood cell count employing an optical disc and disc drive, said method comprising the steps of:
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providing a blood sample in a first tube, the first tube containing a separation gradient;
rotating the first tube at a time and speed sufficient to separate the blood sample into layers;
isolating a white blood cell layer from the separated blood sample;
resuspending the white blood cell layer thereby forming a white blood cell suspension;
providing a sample of the white blood cell suspension on an optical disc surface, the surface including at least one capture zone with at least one capture agent;
loading the optical disc into an optical reader;
rotating the optical disc;
directing an incident beam of electromagnetic radiation to a capture zone;
detecting a beam of electromagnetic radiation formed after interacting with the disc at the capture zone;
converting the detected beam into an output signal; and
analyzing the output signal to extract therefrom information relating to the morphology of the nuclei of the cells captured at the capture zone. - View Dependent Claims (4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
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18. A method of making an optical assay disc for performing a differential white blood cell count using nucleus morphology to distinguish cell types, said method comprising the steps of:
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providing a substrate;
coating said substrate with an active layer;
providing a cross-linker on said active layer to thereby create one or more capture zones;
allowing said cross-linker to bind to said active layer;
removing excess cross-linker from the capture zones; and
attaching a cap portion to said active layer to form a channel adapted to receive a suspension of cells and a dye solution for staining the nuclei of captured cells. - View Dependent Claims (36, 37, 38, 39, 40)
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19. A method of analyzing white blood cells in a test sample, said method comprising the steps of:
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loading a plurality of white blood cells into an optical bio-disc;
capturing said white blood cells in designated target zones by use of capture agents having specificity for particular cell surface markers;
staining the nuclei of the captured white blood cells with a dye;
directing an incident beam of light toward said captured white blood cells;
allowing said incident beam of light to interact with the stained nuclei of the captured white blood cells to thereby form a return beam of light carrying information regarding the morphology of the nuclei;
detecting said return beam of light;
converting said detected return beam into an output signal; and
analyzing said output signal to extract therefrom information relating to the morphology of the nuclei of the cells captured at the capture zone. - View Dependent Claims (20, 21, 22, 23)
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24. A method of conducting an assay employing an optical disc and disc drive, said method comprising the steps of:
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providing a sample of cells on a disc surface;
loading the disc into an optical reader;
rotating the optical disc;
directing an incident beam of electromagnetic radiation to the capture zone;
detecting a beam of electromagnetic radiation formed after interacting with the disc and the sample of cells on the disc surface;
converting the detected beam into an output signal; and
analyzing the output signal to extract therefrom information relating to the number and type of cells captured at the capture zone. - View Dependent Claims (25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35)
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Specification