Confocal Training Information/User Tips
- If any experiment (such as treatment to tissue culture cells) is necessary for the live imaging, this should be performed by the lab member
- Bring your own supplies (pipette, tips, and tissue culture media) for the experiment. If confocal supplies (from Ikezu lab) are consumed, it will be charged to the laboratory
- Strongly recommend following the standard operational procedures(SOPs) for representative experiments, which should contain multiple negative/positive controls. (Experiments without sufficient controls are often a waste of time and effort)
- For more information, please contact the confocal manager: LeAnn Tiede - 402-559,5509 - ltiede@unmc.edu
Guidelines for Sample Preparation
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Limitations
- If the Signal-to-Noise (S/N) ratio is less than 2, it is difficult to reconstruct acceptable images
- If the excitation/emission wavelength does not correspond to the laser and filter specifications, it is difficult to obtain maximal intensity from the sample
- If sample preparation is not adequate (excess mounting media, inappropriate coverglass thickness, inadequate support material), it is difficulty to capture images due to specific focusing depth.
Standard Operational Procedures
- Basic laser scanning confocal imaging
- Z-stack for 3D reconstruction and deconvolution
- Time-course experiment Multiple-positioning experiment
- Fluo-4 Ca2+ imaging
- Fura-2 Ca2+ imaging
- CFP/YFP FRET imaging using Dual/Quadview system
Confocal SOP - pdf file
User Fees* (effective September 1, 2009)
| UNMC** | |
| Consultation/basic training | Free |
| Confocal use (untrained user) | $30/hr |
| Confocal use (trained user) | $15/hr |
| Time lapse studies | $10/hr |
| Time lapse studies (brightfield only) | $5/hr |
| Processing/image analysis | $10/hr |
| Unassisted image analysis | Free |
| Supply costs | variable |
* All fees will be prorated to the nearest half hour
** member of the Department of Pharmacology and Experimental Neuroscience
