Nikon Deconvolution Wide-Field Epifluorescence System
This system is good for:
- General fluorescence fixed and live cell imaging.
- Imaging of weakly fluorescent samples. Time-lapse live-cell imaging.
This system is not good for:
- Very high Z axis resolution
- Fast bleaching dyes
Function of Instrument
This system uses a broadband mercury-halide light source and sets of high-throughput filters to excite fluorescent dyes and proteins.
At present we have 4 filter sets including:
- UV excitation and blue emission for DAPI, Hoechst, and Alexa 350
- Blue excitation green emission for FITC, GFP, and Alexa 488
- Yellow-green excitation red emission for Texas RED, Alexa 594, and mcherry
- Red excitation far red emission Alexa 647 (cy5)
The system has motorized filter changer, nosepiece and focusing drive which allows for automated image acquisition. All imaging parameters are stored in the saved files for later analysis. The focus drive allows for stacks of images at different focal planes to be taken and that information is used to remove out of focus fluorescence and confocal like images to be acquired (deconvolution). The imaging device is a cooled sCMOS camera. This system is well suited for low intensity fluorescence samples.
Training, Usage and Maintenance
- Training involves proper turn on and turn off sequences.
- It also involves learning the proper operation and cleaning so as not to damage the optics or other parts of the instrument.
- Instruction in the use of NIS Elements software is fairly straightforward.
- Generally, the instructor is operating the instrument for the first session and explaining as the experiment progresses and is present for the next experiment if there are questions.
- Maintenance involves cleaning the optics, adding and removing filter cubes for particular experiments and changing the mercury halide lamp.
- New users should schedule a training session of 1 hour. Please book your training 2 weeks in advance. Bring your sample to training and we will acquire images with you.
Help us help you.
Grants: The Cellular Imaging Core operates at a loss and is subsidized by the Department of Medicine. What does not get included in the balance sheets are the indirect costs generated from grants obtained with the help of data from our core. You can help us continue to serve you by letting us know when you submit a proposal and/or are awarded a grant which contains data obtained from the use of our Core.
Acknowledgments: We would greatly appreciate it if authors would acknowledge the Cellular Imaging Core in their publications containing data obtained with the equipment and/or assistance of Core personnel.
Location: 670 Albany Street, EBRC Building – Basement B15.