Method and apparatus for performing contactless optically-induced dielectrophoresis for separation of circulating tumor cells

• US 10,456,791 B2
• Filed: 12/01/2016
• Issued: 10/29/2019
• Est. Priority Date: 04/19/2016
• Status: Active Grant

• Current Assignee: Ace Medical Technology Co., Ltd.
• Original Assignee: Ace Medical Technology Co., Ltd.
• Inventors: Wu, Min-Hsien, Hsieh, Chia-Hsun, Wang, Hung-Ming, Chou, Wen-Pin, Chiu, Tzu-Keng
• Primary Examiner(s): Crawford, Erik B
• Application Number:
  US15/366,593
• Publication Number:
  US 20170297036A1
• Time in Patent Office: 1,062 Days
• Field of Search: None
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0652 Sorting or classification of particles or molecules
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2400/0424 Dielectrophoretic forces
• 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: B03C 2201/26 for use in medical applications
• CPC Class Codes: B03C 5/005 Dielectrophoresis, i.e. dielectric particles migrating towards the region of highest field strength B03C5/02 takes precedence; electrophoresis B01D57/02
• CPC Class Codes: B03C 5/026 using open-gradient differential dielectric separation, i.e. using electrodes of special shapes for non-uniform field creation, e.g. Fluid Integrated Circuit [FIC]
• CPC Class Codes: C12N 13/00 Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
• CPC Class Codes: G01N 15/149 specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: G01N 2001/4038 electric methods, e.g. electromigration, electrophoresis, ionisation
• CPC Class Codes: G01N 2015/1028 Sorting particles

Abstract

A method for performing contactless ODEP for separation of CTCs is provided with the steps of obtaining patients'"'"' blood with rare cell suspected CTCs; adding at least one fluorescent antibody binding to CTCs into the blood; staining the blood; injecting the stained blood with fluorescent dye into an ODEP device and then performing fluorescent image identification; trapping the CTCs with at least one fluorescent antibody in the ODEP device by creating an image pattern and then generating an ODEP force; Separating the trapped CTCs from other non-CTCs cells; absorbing the trapped CTCs; and obtaining a high purity of CTCs. An apparatus for performing contactless ODEP for separation of CTCs is also provided.

Claims

1. A method for performing contactless optically-induced dielectrophoresis (ODEP) for separation of circulating tumor cells (CTCs), the method comprising the steps of:

(1) obtaining blood with suspected CTCs;

(2) adding at least one fluorescent antibody that binds to the CTCs into the blood to form stained blood;

(3) injecting the stained blood with the at least one fluorescent antibody into a sample receiving member of an ODEP device wherein the ODEP device includes a first conductive glass and a second conductive glass, the first conductive glass includes a transverse main channel and a longitudinal micro channel joining the main channel at a cell separation zone, and the first conductive glass includes a first hole and a second hole aligned with two ends of the main channel respectively, a third hole aligned with one end of the microchannel that is distal to the cell separation zone, the sample receiving member disposed on and aligned with the first hole, an exhaust discharge member disposed on and aligned with the second hole, a target collection member disposed on and aligned with the third hole, and a controller including an optical projection device and an image fetch device;

(4) performing fluorescent image identification by immunofluorescent microscopic observation using the optical projection device and the image fetch device to identify and distinguish the CTCs bound to the at least one fluorescent antibody from pseudo positive target cells that have fluorescent expression and lack cell nuclei;

(5) trapping the identified CTCs bound to the at least one fluorescent antibody in the ODEP device by creating an image pattern by projecting a light image and then generating an ODEP force;

(6) separating the trapped CTCs from other non-CTCs at the cell separation zone;

(7) collecting the trapped CTCs in the target collection member; and

(8) obtaining the collected CTCs from the target collection member.

2. The method of claim 1, wherein red blood cells (RBCs) and white blood cells (WBCs) are removed from the blood between step (1) and step (2).