Chronic inflammation can lead to several diseases, thus analysis of the inflammation state of live cells may have important clinical applications. However, there is not currently well-established method for specific detection of inflamed cells via live cell imaging. In this study, we developed an effective antibody (Ab)-based cell imaging method for the detection of inflamed cells using Ab-conjugated nano-beads. Several receptors were tested as potential biomarkers for cell inflammation, and corresponding fluorescence-labeled Abs and/or Ab-conjugated nano-beads were used to detect inflamed cells via fluorescence imaging. Interestingly, when we employed sequential use of TLR1 primary Ab and size-optimized nano-beads conjugated with secondary Ab, we were able to clearly discriminate inflamed cells from normal ones. The Ab-based cell-imaging method described herein provides an important basis for the development of high-throughput analysis of cell inflammation, potentially leading to the identification of factors involved in inflammation and anti-inflammatory drug candidates. (c) 2013 Elsevier Inc. All rights reserved.