Live-cell labeling using CellMask™ Deep Red plasma membrane stain.

Live-cell labeling using CellMask™ Deep Red plasma membrane stain. HaCaT cells (an immortalized human keratinocyte cell line) transfected with a mitochondrion-targeted eGFP construct were incubated for 15 minutes in a solution containing 1 µM Hoechst 33342 (Cat. no. H21492) and then for 5 minutes in a fresh solution containing 8 µg/ml CellMask™ Deep Red plasma membrane stain (Cat. no. C10046); staining took place at 37°C in the presence of 5% CO2. After washing twice with DMEM (including 10% FBS) at 37°C, the cells were imaged with a Zeiss Cell Observer HS microscope fitted with a 100× oil alpha Plan-Apochromat objective and a BFP/GFP/HcRed filter set from Zeiss (62HE). The image stack was deconvolved with the Zeiss AxioVision 3D Deconvolution module with the point spread function acquired using a 200 nm TetraSpeck™ fluorescent microsphere (Cat. no. T7280). Metaphase chromosomes (pseudocolored blue, Hoechst 33342), plasma membrane (pseudocolored red, CellMask™ Deep Red plasma membrane stain), and mitochondria (pseudocolored green, eGFP) are all clearly visible. Image submitted by Christian Junker, University of Saarland, Department of Biophysics, Homburg, Germany.

Live-cell labeling using CellMask™ Deep Red plasma membrane stain. HaCaT cells (an immortalized human keratinocyte cell line) transfected with a mitochondrion-targeted eGFP construct were incubated for 15 minutes in a solution containing 1 µM Hoechst 33342 (Cat. no. H21492) and then for 5 minutes in a fresh solution containing 8 µg/ml CellMask™ Deep Red plasma membrane stain (Cat. no. C10046); staining took place at 37°C in the presence of 5% CO2. After washing twice with DMEM (including 10% FBS) at 37°C, the cells were imaged with a Zeiss Cell Observer HS microscope fitted with a 100× oil alpha Plan-Apochromat objective and a BFP/GFP/HcRed filter set from Zeiss (62HE). The image stack was deconvolved with the Zeiss AxioVision 3D Deconvolution module with the point spread function acquired using a 200 nm TetraSpeck™ fluorescent microsphere (Cat. no. T7280). Metaphase chromosomes (pseudocolored blue, Hoechst 33342), plasma membrane (pseudocolored red, CellMask™ Deep Red plasma membrane stain), and mitochondria (pseudocolored green, eGFP) are all clearly visible. Image submitted by Christian Junker, University of Saarland, Department of Biophysics, Homburg, Germany.

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