Protocol using lentivirus to establish THP-1 suspension cell lines for immunostaining and confocal microscopy

Summary: THP-1, a monocyte cell line growing in suspension, is widely used in immunology research. However, establishing suspension cell lines and performing confocal microscopy can be challenging. Here, we present a protocol to efficiently generate THP-1 cell lines using lentivirus and perform immunostaining and confocal microscopy. We detail steps for virus production, THP-1 cell infection and clone selection, fixing the suspension cells to the glass slide for immunostaining, and subsequent confocal microscopy. This protocol can be applied to other suspension cells.For complete details on the use and execution of this protocol, please refer to Ji et al. (2021).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.