The ImageStream system can quantify the intensity, specific location, and distribution of signals within tens of thousands of cells per sample. The best applications take advantage of the technology's imaging capabilities to discriminate subtle morphologic or signal distribution changes within individual cells and cell populations.
The available offline IDEAS software (ver 6.2) has pre-configured and optimized wizards for many common applications.
Lasers: 405nm, 488nm, 561nm, 642nm, 785nm (for SSC)
Objective: 40x 0.7 NA
|Services with operator assistance
|Services without operator assistance (must be trained)
|Cancellation Charges (less than 24-hour notice)
For online scheduling navigate to iLabs for the core login page: https://ouhsc.corefacilities.org/
Minimum appointment time is 15 minutes and anything longer than 4 hours needs approval.
Any problems with iLab calendar read the help manuals below:
WHAT YOU NEED TO BRING
- Final Sample Concentration and Volume: At least 1 million cells in 50 ul (2x107 cells/ml) in PBS/2%FVS in a 1.5ml siliconized micro centrifuge tube. Will run ~400 cells per second on low speed.
- Protocols: In general, any established labeling protocol used for flow cytometry will work the ImageStream (see Current Protocols in Cytometry for general labeling techniques). Stain cells on ice in the presence of azide when possible to reduce non-specific capping of antibody. Use siliconized polypropylene tubes when possible.
- Choice of Fluorochromes: Choose fluorochromes that are excited by the lasers in your ImageStream (405, 488, 561, 642nm). Use spectra viewer online to help you plan which dyes will work best.
- Compensation: Have a sample of cells each labeled with a single-color for each fluorochrome used (i.e. FITC only cells, PE only cels, etc.).
- Cell Agggregration: Minimize aggregation problems by straining the sample through a 40um nylon mesh strainer, or by using and anti-clumping buffer such as EDTA or Accumax prior to fixation.
- Fixation: If fixation is desired, thoroughly fix cells with 1% PFA on ice for 20 min.
- Number of samples: No more than 30 total for feasibility experiments. PLEASE LIMIT THE SAMPLES TO THE FOLLOWING; Positive and Negative biologic controls, compensation controls, and experimental samples.
- Lab coat
SAFE OPERATING PRACTICES (read before using equipment)
SOP for ImageStream