BD PharmingenTM Hoechst 34580

Description

Image analysis of live HeLa cells stained with Hoechst 34580. Live cells from the human HeLa Cervical adenocarcinoma, ATCC CCL-2 cell line were stained with 5 mug/mL BD Pharmingen Hoechst 34580 Cat. No. 565877 to segment nuclei pseudocolored blue , 200 nM BD Pharmingen MitoStatus Red Cat. No. 564697 pseudocolored red to stain polarized mitochondria, and 10 muM Calcein AM Cat. No. 564061, pseudocolored green for cytoplasmic staining. The image was captured using a Molecular Devices ImageXpress Micro XLS with a 20 objective and merged using Molecular Devices MetaXPress software. Flow cytometric analysis of HeLa cell DNA content using Hoechst 34580 staining. HeLa cells in log phase growth were dissociated using 0.25 % Trypsin-EDTA Life Technologies . Cells were resuspended at 1 106 cells/mL in culture medium containing 1.5 mug/mL BD Pharmingen Hoechst 34580 and stained 30 minutes, 37C . Cells were pelleted, resuspended in DPBS, and analyzed by flow cytometry using a low flow rate. A 405 nm laser with a 450/50 bandpass filter was used to collect data; comparable results have also been obtained using a 355 nm laser with a 450/50 bandpass filter or a 405 nm laser with a 470/100 bandpass filter. Data was acquired using a BD LSRFortessa Cell Analyzer System. Histograms showing DNA levels were deconvoluted by FlowJo software FlowJo, LLC into G0/G1, S, and G2/M populations green lines . Image analysis of live HeLa cells stained with Hoechst 34580. Live cells from the human HeLa Cervical adenocarcinoma, ATCC CCL-2 cell line were stained with 5 mug/mL BD Pharmingen Hoechst 34580 Cat. No. 565877 to segment nuclei pseudocolored blue , 200 nM BD Pharmingen MitoStatus Red Cat. No. 564697 pseudocolored red to stain polarized mitochondria, and 10 muM Calcein AM Cat. No. 564061, pseudocolored green for cytoplasmic staining. The image was captured using a Molecular Devices ImageXpress Micro XLS with a 20 objective and merged using Molecular Devices MetaXPress software. Show More Flow cytometric analysis of HeLa cell DNA content using Hoechst 34580 staining. HeLa cells in log phase growth were dissociated using 0.25 % Trypsin-EDTA Life Technologies . Cells were resuspended at 1 106 cells/mL in culture medium containing 1.5 mug/mL BD Pharmingen Hoechst 34580 and stained 30 minutes, 37C . Cells were pelleted, resuspended in DPBS, and analyzed by flow cytometry using a low flow rate. A 405 nm laser with a 450/50 bandpass filter was used to collect data; comparable results have also been obtained using a 355 nm laser with a 450/50 bandpass filter or a 405 nm laser with a 470/100 bandpass filter. Data was acquired using a BD LSRFortessa Cell Analyzer System. Histograms showing DNA levels were deconvoluted by FlowJo software FlowJo, LLC into G0/G1, S, and G2/M populations green lines . Show More Image analysis of live HeLa cells stained with Hoechst 34580. Live cells from the human HeLa Cervical adenocarcinoma, ATCC CCL-2 cell line were stained with 5 mug/mL BD Pharmingen Hoechst 34580 Cat. No. 565877 to segment nuclei pseudocolored blue , 200 nM BD Pharmingen MitoStatus Red Cat. No. 564697 pseudocolored red to stain polarized mitochondria, and 10 muM Calcein AM Cat. No. 564061, pseudocolored green for cytoplasmic staining. The image was captured using a Molecular Devices ImageXpress Micro XLS with a 20 objective and merged using Molecular Devices MetaXPress software. Flow cytometric analysis of HeLa cell DNA content using Hoechst 34580 staining. HeLa cells in log phase growth were dissociated using 0.25 % Trypsin-EDTA Life Technologies . Cells were resuspended at 1 106 cells/mL in culture medium containing 1.5 mug/mL BD Pharmingen Hoechst 34580 and stained 30 minutes, 37C . Cells were pelleted, resuspended in DPBS, and analyzed by flow cytometry using a low flow rate. A 405 nm laser with a 450/50 bandpass filter was used to collect data; comparable results have also been obtained using a 355 nm laser with a 450/50 bandpass filter or a 405 nm laser with a 470/100 bandpass filter. Data was acquired using a BD LSRFortessa Cell Analyzer System. Histograms showing DNA levels were deconvoluted by FlowJo software FlowJo, LLC into G0/G1, S, and G2/M populations green lines .