Confocal and fluorescent microscopes provide digital images of specific molecules and organelles within cells and the images are processed by computers. These methods are essential tools for modern research in cell biology and methods of cell and molecular biology are the most widely used methods for basic research performed in RCER. Digital imaging is based on computer manipulation of digital data and uses involved methods of data processing.
Rammelkamp Core Facilities have a new Leica TCS SP2 Confocal system. This system employs new technology involving a spectrophotometer detection system instead of the traditional filter system. The user will be able to change the bandwidth of the emission spectrum so that crosstalk between fluorochromes can be minimized. It will also provide more flexibility as new fluorochromes become available. This confocal microscope is equipped with a Argon/Krypton Laser that will excite at 457nm, 488nm, and 568nm wavelengths and a red HeNe laser that will excite at 633nm. This system is able to obtain high resolution images of tissue section or cells. The Core Facilities also includes a Noran Oz Confocal system with an Argon laser (excites at 488 and 527nm) is mounted on a Nikon inverted microscope. This system is used to obtain high resolution fluorescent images of tissue slices and cultured cells. 2-D and 3-D reconstruction software available. Both confocal systems are curently available as a service or with training it can be can be used independently. A fee will be charged for usage.
Olympus BH-2 upright microscope with SPOT digital camera and 20x, 40x and 100x objectives is equipped for normal light or fluorescent images. Filter sets for DAPI, fluorescein and rhodamine. Training required and then available on sign-up. A Nikon E600 upright microscope has been added to the facility. It has 4x, 10x, 20x, 40x, 100x objectives, as well as a 40x water immersion lens. The fluorescent filters are for DAPI, FITC, RITC and Texas Red. Additionally, this microscope is equipped for phase contrast at 20x, and DIC at 40x. A Spot Digital Camera is attached to the Nikon E600 microscope to capture fluorescent and brightfield images. Training on the microscopes and camera is required and equipment is available by reservation.
The SLM 8100 Spectrofluorometer can be used for a variety of fluorescence measurements including real-time intracellular ion concentration measurements using ratiometric fluorescent probes. It has multiple excitation and emission wavelength capability, temperature controlled cuvette chamber, and magnetic stirring device. Software programs for data acquisition and analysis are an integral part of the system and readily available for all users.