Paper Title
Crosstalk-Eliminated Quantitative Determination of Aflatoxin B1-Induced Cancer Stem Cells in Hepatocellular Cancer

Abstract
In this work, it was found that hepatocellular cancer stem cell (CSCs) were produced from HepG2 cells by aflatoxin B1-induced mutation and their amount were quantitatively determined using a crosstalk-eliminated multicolor cellular imaging based on quantum dot (Qdot) nanoprobes and an acousto-optical tunable filter. Hepatocelluar CSCs were acquired from HepG2 cells via magnetic bead-based sorting and observed using concurrent detection of three different markers CD133, CD44 and aldehyde dehydrogenase1 (ALDH1). The DNA mutation of HepG2 cells caused by aflatoxin B1 was quantitatively observed via absorbance spectra of aflatoxin B1-8,9-epoxide-DNA adducts formed in HepG2 cells by the treatment of HepG2 cells with aflatoxin B1. The number percentages of hepatocellular CSCs formed in the entire HepG2 cells were determined to be 9.77±0.65%, 10.9±1.39%, 11.4±1.32%, and 12.8±0.7%, respectively at the control, 5μM, 10μM, and 20μM aflatoxin B1. The results were well matched with those obtained using flow cytometry used as a general tool for the detection of hepatocellular CSCs. This study demonstrates that accurate quantitative measurement of aflatoxin B1-induced hepatocellular CSCs can be accomplished using a constructed multicolor cellular imaging system that eliminates the crosstalk among hepatocellular CSC biomarkers. Index Terms - Aflatoxin B1, cancer stem cell, quantum dot, multicolor cellular imaging.