Resources
- DNA Content for Cell Cycle Analysis of Fixed Cells With Propidium Iodide
- Apoptosis Assays (Sub Diploid, Annexin V, TUNNEL, Caspase Activity)
- Simultaneous analysis of DNA content and surface immunophenotype using gentle ethanol fixation techniques.
- CFSE Labeling
- Intracellular Cytokine Staining Protocol
- Measurement of GFP Expression and DNA Content in Permeabilized Cells
- 7-AAD Staining of Dead Cells
- Critical Aspects of Staining Cells for Flow Cytometry
- Preparation of 2% Formaldehyde Solution
- Fluorochrome Conjugation of Monoclonal Antibodies
- Staining Protocol for the Cell Permeant DNA Dye DRAQ5
- Reducing Cellular Autofluorescence
- Dead Cell Discrimination by EMA
- Protocol for LPS Activation of MAPK Signaling in Whole Blood Samples
- Protocol for Antibody Titration
- Mass Cytometry Methods and Protocols
- Immunophenotyping of Human Peripheral Blood Mononuclear Cells by Mass Cytometry (Heck S, Bishop CJ, Ellis RJMethods Mol Biol. 2019, 1979, 285-303)
- Preparing Viable Single Cells from Human Tissue and Tumors for Cytomic Analysis (Leelatian N, Doxie DB, Greenplate AR, Sinnaeve J, Ihrie RA, Irish JM. Curr Protoc Mol Biol. 2017)
- Phenotyping of Live Human PBMC using CyTOF Mass Cytometry (Leipold and Maecker 2015)
- Intracellular Cytokine Staining on PBMCs Using CyTOF Mass Cytometry (Lin, Gupta, and Maecker 2015)
- Multiparameter Phenotyping of Human PBMCs Using Mass Cytometry
- A Deep Profiler’s Guide to Cytometry
- Mass Cytometry Selected Publications
- DVS Sciences – Metal-Conjugated Antibodies (anti-Human)
- Panel Design Guidelines
- Experiment Planning Worksheet
- Cell Surface Staining Protocol
- Cytoplasmic Staining Protocol
- Intercalator Ir-125 Protocol
- Intercalator Rh500 uM (Live Dead) Protocol
- Phospho-Protein Staining Protocol
- Nolan Lab Signaling based Resource (Fluorescence & Mass)
- BD Buffer/Fixative Resource
NIH Recommendations for Rigorous Resources for Rigorous Research
ScientificScientific Premise in NIH Grant Applications
Enhancing Reproducibility through Rigor and Transparency
Clearinghouse for Training Modules to Enhance Data Reproducibility
Click here for training modules
Below are the links to Optimized Multicolor Immunophenotyping Panels (OMIPs). These panels have been validated and published and are a useful resource for developing immunophenotyping panels for a variety of applications.
The following article contains background and explanation as the development of the OMIPs as a separate type of publication within the Cytometry Part A journal:
http://onlinelibrary.wiley.com/doi/10.1002/cyto.a.20916/pdf
Useful Links
FlowRepository.org is a Web-based flow cytometry data repository provided by the International Society for Advancement of Cytometry (ISAC). It supports storage, annotation, analysis, and sharing of flow cytometry datasets. A fundamental tenet of scientific research is that published results should be open to independent validation and refutation. With FlowRepository, researchers can annotate their datasets in compliance with the Minimum Information about a Flow Cytometry Experiment (MIFlowCyt) standard, thus greatly facilitating third-party interpretation of their data.
Tools and Tutorials
- BenchSci
- FCS Express Tutorial Videos
- Fluorofinder 2.0 Flow Cytometry Panel Builder
- Purdue University Cytometry Laboratories
- Intracellular Cytokine Staining Protocol
- Counting Statistics – How Many Cells Do I Collect?
- Practical Flow Cytometry (4th edition-Free Online Viewing)
- Beginner Flow Cytometry Book (Flow Cytometry A basic introduction)
Flow Cytometry Vendor Links
- BD Biosciences (reagents) Susan Gigler ([email protected])
- Beckman Coulter (reagents)
- BioLegend (reagents) Hank Beggs ([email protected])
- Bio-Techne Ty Vaivoda ([email protected])
- Cytek Biosciences (reagents)
- Luminex (Amnis)
- Miltenyi Biotec (reagents) Christina Stillwell ([email protected])
- Phoenix Flow (Accutase/reagents/software)
- Spherotech (Beads)
- Thermo Fisher Scientific (Caltag Reagents, Molecular Probes Reagents, Quantum Dot Reagents, Dynal Beads) Alycia Curlee ([email protected])