Protocols & Reagents

The following PDF contains useful information about the facility and sample preparation:

• FCCFinfo

Please fill out and submit the following forms when submitting a new project or using the Aria II (Athena):

• Project description form
• IBC/Biohazard form (if sample has an IBC protocol and/or is a biohazard)
• Aria II analysis or sorting form

These protocols have been used by researchers at IU Bloomington:

• Apoptosis detection – Annexin V-AF488 vs PI (Thermo Fisher)
• Cell Cycle – Propidium Iodide (UCSD) - add ethanol to cells
• Cell Proliferation (thermo Fisher)
• Drosophila Embyro sorting (pdf)
  
  

Useful guides:

• Antibody titration
• Sorting Guidelines (CCHMC RFCC)
  
  

Additional assays and sorting performed by researchers at IU Bloomington:

• Surface marker Staining and sorting (Stem & Adherent cells - BD Biosciences)
• Intracellular marker staining (BioLegend)
• Measurement of Green Fluorescent Protein (GFP) expression and sorting
• Single Cell Sorting
• Mitochondrial membrane potential (thermoFisher)
• Viability assays (Live/Dead dyes - BioLegend)
• Bacterial viability & counting (Viability; Viability & counting - Thermo Fisher)
  
  
• Protocol for Sulfolobus fixation and cell cycle analysis available through IUCAT - Analysis of the Archaeal ESCRT Apparatus
  
  

Other assays available:

• BrdU incorporation
• Cell proliferation assays
• Intracellular Cytokine detection
• Multiparameter Immunofluorescence analysis and cell sorting
• Signal Transduction
• and much more!

• Current Protocols in Cytometry (Facility has a copy on permanent loan from Libraries)   
• Molecular Probes Handbook
• Cytometry Part A (Journal)
• UCSD Protocols Page
• BD Biosciences
• ThermoFisher protocols
• COPAS Select, Union Biometrica - search protocols
• Flow Cytometry with Plant Cells (Wiley, E-Book, IU-B Life Sciences Library)