The Imaging and Morphology Support Core of the ONPRC is designed to meet the imaging needs of ONPRC scientists and of the OHSU scientific community by providing state-of-the-art facilities, expertise, assistance and training in the use of advanced light microscopy, image analysis, processing and printing, and laser capture microdissection. Special emphasis is placed on quantitative evaluation of imaging experiments using both stereology and automated image analysis.
Workstation has motorized stage, DG4 fluorescence illumination and the stereology, FURA and FRET packages.
Has UV cutting and fluorescence capability.
Spectral confocal system with 5 excitation lasers (405, 458, 476, 488, 514, 561, 594, 633nm), motorized stage and stage-top microincubator.
Microscope has pulsed IR laser for 2-photon excitation in addition to 3 visible light lasers.
"Fluorescence recovery after photobleaching (FRAP) denotes an optical technique capable of quantifying the two dimensional lateral diffusion of a molecularly thin film containing fluorescently labeled probes, or to examine single cells. This technique is very useful in biological studies of cell membrane diffusion and protein binding. In addition, surface deposition of a fluorescing phospholipid bilayer (or monolayer) allows the characterization of hydrophilic (or hydrophobic) surfaces in terms of surface structure and free energy. Similar, though less well known, techniques have been developed to investigate the 3-dimensional diffusion and binding of molecules inside the cell; they are also referred to as FRAP."
"Fluorescence resonance energy transfer (FRET) is a distance-dependent interaction between the electronic excited states of two dye molecules in which excitation is transferred from a donor molecule to an acceptor molecule without emission of a photon. The efficiency of FRET is dependent on the inverse sixth power of the intermolecular separation, making it useful over distances comparable to the dimensions of biological macromolecules. Thus, FRET is an important technique for investigating a variety of biological phenomena that produce changes in molecular proximity. When FRET is used as a contrast mechanism, colocalization of proteins and other molecules can be imaged with spatial resolution beyond the limits of conventional optical microscopy."
"Fura-2, a polyamino carboxylic acid, is a ratiometric fluorescent dye which binds to free intracellular calcium. It was the first widely-used dye for calcium imaging, and remains very popular. Fura-2 is excited at 340 nm and 380 nm of light, and the ratio of the emissions at those wavelengths is directly correlated to the amount of intracellular calcium. The use of the ratio automatically cancels out confounding variables, such as variable dye concentration and cell thickness, making Fura-2 one of the most appreciated tools to quantify calcium levels. More recently, genetically-encoded calcium indicators based on spectral variants of the green fluorescent protein, such as Cameleons, have supplemented the use of Fura-2 and other small molecule dyes for calcium imaging."
"Time-lapse microscopy (microphotography, photomicrography) - microscopy in which the same object (e.g., a cell) is photographed at regular time intervals over several hours.
Live-cell imaging often involves time-lapse microscopy to monitor cell movements. Modern approaches are extending these observations beyond making movies of cell structure. Increasingly, time-lapse imaging is being integrated with specialized techniques for monitoring, measuring, and perturbing dynamic activities of cells and subcellular structure"
Includes Fura-2 calcium imaging, FRET, FRAP imaging. Consultation, training and access to instruments or full support by core personnel are offered for all services.
"ImageJ is a public domain Java image processing program inspired by NIH Image for the Macintosh. It runs, either as an online applet or as a downloadable application, on any computer with a Java 1.4 or later virtual machine. Downloadable distributions are available for Windows, Mac OS, Mac OS X and Linux.
It can display, edit, analyze, process, save and print 8-bit, 16-bit and 32-bit images. It can read many image formats including TIFF, GIF, JPEG, BMP, DICOM, FITS and "raw". It supports "stacks", a series of images that share a single window. It is multithreaded, so time-consuming operations such as image file reading can be performed in parallel with other operations.
It can calculate area and pixel value statistics of user-defined selections. It can measure distances and angles. It can create density histograms and line profile plots. It supports standard image processing functions such as contrast manipulation, sharpening, smoothing, edge detection and median filtering."
"Neurolucida is advanced scientific software for brain mapping, neuron reconstruction, anatomical mapping, and morphometry."
Whole slide stereology software.
"Comprehensive Digital Microscopy Package: SlideBook software carries biomedical imaging through the entire experimental process. By addressing everything from device control to image processing to automated data analysis, SlideBook allows users to focus on scientific investigation and novel methods rather than on core instrument functionality."
"Stereo Investigator gives you image analysis, mapping, and measurement tools all in one software package, with more than 20 stereological probes, including the Optical Fractionator, Cavalieri, and Nucleator."
"High Performance 3D-4D imaging software for a better insight to your science."