Cells were seeded into 6-well plates at a density of 300,000 cells per well in 2 mL medium for 16 hours. The cells were treated with the indicated drugs and ionizing radiation (IR), and cultured for 48 hours. Cell apoptosis was assessed by Annexin V-FITC (Invitrogen) and propidium iodide (PI; Sigma-Aldrich) staining coupled with flow cytometry analysis. For cell cycle distribution analysis, cells were fixed in 70% ethanol at −20°C and stained with DNA staining solution containing PI and RNaseA (Sigma-Aldrich) overnight. All data were acquired on LSRII cytometry (BD Biosciences) and each sample was assessed using a collection of 10000 events, followed by analysis using FlowJo software (FlowJo, Ashland, Oregon).
Lsrii cytometry
Manufactured by BD
The BD LSRII is a flow cytometry system that enables the analysis of multiple parameters on single cells or particles. It can detect and measure various physical and fluorescent characteristics of cells or other particles as they pass through a laser beam, providing quantitative data on their properties.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using lsrii cytometry
1
Cell Apoptosis and Cycle Analysis
2
Mitochondrial Oxidative Stress Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
As described by our previous studies [19 (link),22 (link)], the mitochondrial oxidative stress assay was performed using MitoSOX Red dye (Invitrogen, Burlington, ON, Canada), which was detectable by flow cytometry. Briefly, Ca9-22 cells were treated with single drugs or by a combination of cisplatin and PAC for 24 h. After the treatment, 5 µM of MitoSOX™ Red was added to each sample (106 cells/mL) and incubated in the dark 1 h at 37 °C. After incubation, the cells were washed twice with a warm buffer before analyzing by flow cytometry at 510 nm using LSRII cytometry from BD Bioscience.
3
Evaluating Cell Response to AZD1775 and Ionizing Radiation
4
Immune Cell Profiling in Mediastinal Lymph Nodes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mediastinal lymph nodes and lungs were harvested at indicated days of infections and processed to generate single cell suspension as previously described 12, 55 . A list of all antibodies used for ow cytometery in this study is in Supplemental Table 2. In general, cells were rst stained with surface markers at 30 min room temperature (RT) and an additional 30 min on ice. Cells were then xed and permeabilized with Foxp3 x/perm (Tonbo) before being stained for intracellular markers for 30 min on ice. For OTII TCR speci c tetramer staining, cells were incubated with I-A(b) AAHAEINEA tetramers in 50µl for 1hr in 37ºC incubator before addition of other surface markers with additional 30 min at RT before xation as described above. For Annexin V staining, cells were stained for surface markers and washed before staining for Annexin V for 20 min at RT. Cells were washed and quickly analyzed in a cytometer. All ow cytometry samples were acquired using LSRII cytometry (BD) with FACSDiva software. All cell sorting were performed in BD In ux cell sorter. Data were analyzed using FSC Express (DeNovo software).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!