Live Cell Imaging of Hypoxia in Cancer Cells using the CellASIC® ONIX2 Microfluidic System
A431 epidermal carcinoma cells (85090402) were incubated 16 hours overnight with 3µM BioTracker 520 Green Hypoxia Dye (SCT033) in complete DMEM by gravity perfusion at 37°C under normoxic conditions (20% O2) using the CellASIC® ONIX2 System. After overnight incubation, cells were perfused with complete DMEM media without dye at 1 psi under hypoxic conditions (0% O2) for 16 hours at 37°C. Fluorescent images were captured over 16 hours using 465 nm excitation, and a 525 nm emission filter. The BioTracker 520 Green Hypoxia Dye increases in fluorescence intensity with decreasing oxygen levels (increased hypoxic conditions).
Live Cell Imaging of p62 using LentiBrite™ Fluorescent Biosensors
Human mesenchymal stem cells (MSCs), were plated in a chamber slide and transduced with LentiBrite™ RFP-p62 lentiviral particles at an MOI of 40 for 24 hours. After media replacement and 24 hours further incubation, cells were incubated for 4 hours in EBSS containing a lysosome inhibitor, to induce autophagy and inhibit lysosomal degradation of autophagosomes.
Live Cell Imaging of β-Actin Cytoskeleton Proteins using LentiBrite™ Fluorescent Biosensors
U2OS cells were plated in a chamber slide and transduced with LentiBrite™ GFP-β-actin lentiviral particles at an MOI of 20 for 24 hours. After media replacement and an additional 48 hours of further incubation, the cells were treated with 2 µM cytochalasin-D for another 2 hours, to induce actin depolymerization. The GFP- β -actin displays a filamentous distribution in the untreated cells, and upon actin depolymerization, adopts a punctate distribution. LentiBrite™ Lentiviral Biosensors are a new suite of pre-packaged lentiviral particles encoding important and foundational proteins of autophagy, apoptosis, and cell structure for visualization under different cell/disease states in live cell and in vitro analysis.
Live Cell Imaging of Autophagy using LentiBrite™ Fluorescent Biosensors
HT-1080 cells were plated in a chamber slide and transduced with LentiBrite™ GFP-LC3 lentiviral particles at an MOI of 20 for 24 hours. After media replacement and 48 hours further incubation, cells were incubated for 4 hours in EBSS containing a lysosome inhibitor, to induce autophagy and inhibit lysosomal degradation of autophagosomes. LentiBrite™ Lentiviral Biosensors are a new suite of pre-packaged lentiviral particles encoding important and foundational proteins of autophagy, apoptosis, and cell structure for visualization under different cell/disease states in live cell and in vitro analysis.
Live Cell Imaging of Apoptosis in Immune Cells using the CellASIC® ONIX2 Microfluidic System
Live Cell Imaging of Apoptosis in Immune Cells using the CellASIC® ONIX2 Microfluidic System
Optimized Staining and Proliferation Modeling Methods for Cell Division Monitoring using Cell Tracking Dyes
Fluorescent cell tracking dyes, in combination with flow and image cytometry, are powerful tools with which to study the interactions and fates of different cell types in vitro and in vivo.