Isolation and detection of A549 lung cancer cells based on magnetic separation and indirect immunofluorescence technologies

This article reports the development of a combining immunomagnetic separation technology with indirect immunofluorescence technology for separating and determining of the carcinoma A549 cells.Magnetic beads weresynthesized by chemical precipitation.Immunomagneticbeads were constructed by grafted EGFR to the surface of Fe₃O₄ nanoparticles coated by lipid.The carcinoma A549 cells could specifically bind with the EGFR antibody on the immunomagnetic's surface.The carcinoma A549 cells could be isolatedand enriched from the mixed cellsunder the action of in the magnetic field.The separated cells could connect with the specific antibody contained PE fluorophore and the location of cellnucleus can be confirmed by DAPI to stain the cell nucleus.The carcinoma A549 cells could be quantitatively detectedbyobserving inversion fluorescence microscope.All those results indicate that the method of combining Immunomagnetic separation technology and indirect immunofluorescence technology have a good specificity and high sensitivity for isolationand detection of the carcinoma A549 cells.thedetection limit is three cells per milliliter