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Figure 1 | BMC Research Notes

Figure 1

From: The use of HaloTag-based technology in flow and laser scanning cytometry analysis of live and fixed cells

Figure 1

Laser scanning cytometry analysis of β1-integrin expression and cell cycle staining in the U2OS-β1Int-HT7 cell line. A. Field images for U2OS-β1Int-HT7 cells treated with MJ (1 mM and 0.5 mM) or left untreated for 16 h and then stained with Oregon Green and Alexa 488 HT ligands. Blue is cell nuclei stained with Hoechst 33342; green is the β1 integrin-HT fusion protein stained with Oregon Green HT ligand; red is nuclei of necrotic cells stained with PI. Images were acquired with two lasers (405 and 488 nm). Two types of primary contouring were applied to the samples: blue object image (based on nucleus contouring) and phantom. The same pattern in fluorescence intensity levels was achieved with Oregon Green and Alexa 488 HT ligands. B. Analysis of green cell fluorescence based on phantom primary contouring in samples treated with different doses of STS (0.25 μg or 0.5 μg/ml) or MJ (0.5 or 1 mM), or left untreated. C. The protocol of image acquisition on the laser scanning cytometer. D, E, F. Cell cycle analysis was performed by Hoechst staining. Dot-blots are shown of cell populations gated on the base of blue (D) and red (E) fluorescence. A cell cycle histogram and pre-G1-peak (apoptotic subpopulation) defined on the basis of Hoechst staining (F).

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