Detection of rare cells in a microfluidic disk systemia positive selection
Date Issued
2009
Date
2009
Author(s)
Pan, Yu-Cheng
Abstract
In clinical studies, concentration of specific rare cells in the blood can serve a variety of purposes, e.g. disease diagnostics, evaluation of treatment efficacy, and disease prognosis. With the ratio of rare cells to whole blood cells in the range of one per billion (ppb), detection and separation of rare cells from whole blood via quick and inexpensive methods present important challenges in medicine. This thesis presents a proof-of-concept study via a compact disk (CD)-based microfluidic platform to separate and detect rare cells in blood. The disk platform utilized immuno-beads and a multi-stage magnet to separate rare cells. Detection of rare cells was performed in the disk with. The transfer force of liquid translation was centrifugal force via adjusting the rotational speed. The blood of patients was inconveniently obtained, so the cell lines were used in the preliminary experiments. The MCF7 were used to simulate the circulating tumor cells (CTCs) as target and stained with anti-CD326-PE antibodies and anti-PE BD magnetic beads. Jurkat Clone E6-1 was used to simulate the large number of cells in human blood, such as leukocyte, erythrocyte, and other cells. In order to identify with MCF7, Jurkat were labeled with Calcein-AM. The 300μl cell mixture containing few MCF7 and great number of Jurkat was injected into the microfluidic disk. And then the multi-stage magnet was put at the top of disk for two minutes. After rotation, the MCF7 was captured at the top of inlet reservoir, and Jurkat and solution were depleted to outer rim. The captured MCF7 in mircofluidic disk were detected with a fluorescence microscope. Results indicate that while small amount of MCF7 cells (10+、100+、300+) mixed into three different amount of Jurkat cells (106, 107, 108), the yield of detecting MCF7 were approximately 80%. The yield of MCF7 is independent of the number of Jurkat, and the microfluidic disk has well sensitivity (one in 107). The compact disk performs separation and enumeration of rare cells within roughly 30 minutes with relative ease and low cost at an acceptable level of cell loss. The microfluidic disk should be readily applied to separate and detect many rare cells with distinct immunological features, such as CTCs in cancer patients’ blood, the fetal cells in maternal blood, and endothelial cells in human blood. According to more information of diagnosis, doctor would make the personal treatment for patients. And the microfluidic disk could reduce the risk of assays.
Subjects
rare cell
microfluidics
positive selection
disk
centrifugal force
SDGs
Type
thesis
File(s)![Thumbnail Image]()
Loading...
Name
ntu-98-R96543079-1.pdf
Size
23.53 KB
Format
Adobe PDF
Checksum
(MD5):f8d92fa35a5efb3815bb2231c63396d5