The largest database of trusted experimental protocols

Lsr fortessa flow cytometry machine

Manufactured by BD

The BD-LSR Fortessa is a flow cytometry machine designed for high-performance cell analysis. It is equipped with multiple lasers and detectors, enabling the simultaneous measurement of multiple parameters on individual cells or particles within a sample. The core function of the BD-LSR Fortessa is to rapidly analyze and characterize the physical and fluorescent properties of cells or other particles in a fluid suspension.

Automatically generated - may contain errors

2 protocols using lsr fortessa flow cytometry machine

1

Neutrophil Characterization by Flow Cytometry

Check if the same lab product or an alternative is used in the 5 most similar protocols
Peripheral blood was collected in EDTA vial, and 200 μl of blood was aliquoted for antibody staining. Briefly, 1 ml of RBC lysis buffer was added to 200 μl of whole blood, and mixed properly. A 10× red blood cell lysis buffer was prepared in-house using NH4Cl (0.155 M), KHCO3 (0.01 M) and EDTA (0.1 mM). The tubes were incubated for 10 min at 4 °C, and centrifuged at 300×g for 10 min. The cell pellets were washed twice with 1× PBS and suspended in 300 μl of PBS to obtain a single cell suspension. For cell surface staining, 100 μl of cell suspension was used, and the antibodies CD16 (1:100) (clone 3G8, APC conjugated, Cat No 302011, Biolegend, USA), CD66b (1:100) (clone G10F5, FITC conjugated, Cat No 305103, Biolegend, USA) and CD177 (1.5:100) (clone MEM-166, APC/Cyanine 7 conjugated, Cat No 315809, Biolegend, USA) was used. Unstained controls, stained samples, and fluorescence minus one controls were acquired on BD-LSR Fortessa flow cytometry machine. Using the FACS DIVA software, PMN gating was done based on FSC-A v/s SSC-A plot. Enriched and activated neutrophils were gated based on CD16+ CD66b+ (double positive) in a quadrant plot. CD177+ neutrophils were gated within these double positive cells.
+ Open protocol
+ Expand
2

ADCP Assay for Sabatolimab Activity

Check if the same lab product or an alternative is used in the 5 most similar protocols
Human PBMCs were separated from whole blood (from the internal donor program at Novartis) using Leucosep tubes, and monocytes were isolated following the Miltenyi Biotech Classical Monocyte Isolation Kit. Cells were plated in a 96-well round bottom plate at a concentration of 5 × 104 cells/well and were incubated for 7 days with 50 ng/ml M-CSF at 37°C. On day 7, various concentrations of sabatolimab (or hIgG4) were added to the cells for 10 minutes at RT, followed by addition of 5 × 104 cells (100 μl) of CFSE-labeled target cells (SKM-1 or HNT-34). Plates were incubated for 4 hours at 37°C at 5% CO2. After 4 hours, cells were washed with PBS, stained for CD11c, and run on a BD LSR Fortessa flow cytometry machine. Data were analyzed using FlowJo analysis software and graphed using GraphPad Prism. ADCP activity was determined as percentage of CFSE+CD11c+ events.
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!