Abstract
Super-paramagnetic microbeads are widely used for cell isolation. Evaluation of the binding affinity of microbeads to cells using optical microscopy has been limited by its small scope. Here, magnetic property of microbeads was first investigated by using synchrotron radiation (SR) in-line x-ray phase contrast imaging (PCI). The cell line mouse LLC (Lewis lung carcinoma) was selected for cell adhesion studies. Targeted microbeads were prepared by attaching anti-VEGFR2 (vascular endothelial growth factor receptor-2) antibody to the shell of the microbeads. The bound microbeads were found to better adhere to LLC cells than unbound ones. PCI dynamically and clearly showed the magnetization and demagnetization of microbeads in PE-50 tube. The cells incubated with different types of microbeads were imaged by PCI, which provided clear and real-time visualization of the cell isolation. Therefore, PCI might be considered as a novel and efficient tool for further cell isolation studies.
Highlights
Cell isolation is an important technique for therapy or research
Cell Culture Experiments Immunocytochemical staining for VEGFR2 expression was positive for Lewis lung carcinoma (LLC) cells (Fig. 2)
Our results have suggested that phase contrast imaging (PCI) is a feasible, non-invasive, and real-time tool for imaging cell isolation
Summary
Cell isolation is an important technique for therapy or research. Microbeads are designed for cell isolation with high binding capacity and good magnetic property [1].The specific antibodies conjugated onto the surface of the magnetic microbeads can be used to recognize their target proteins expressed in cells [2,3]. The bound microbeads with specific marker can help detect circulating tumor cells (CTCs) in the blood or body fluids. The high-affinity binding of microbeads enhances the sensitivity of cancer cell detection. It is very important to estimate the binding affinity of the bound microbeads to their target before their clinical application. Optical microscopy is often used to figure out the magnetic beads and clarify their capability in cell isolation; the imaging method could only be used for observing the surface of the specimens. The field of view for optical microscopy is relatively narrow
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