Penn Dental Medicine houses a Radiance 2100 laser confocal microscope available for use by researchers throughout the School as well as others inside and outside the University.
"Penn Dental Medicine houses a Radiance 2100 laser confocal microscope with argon, green He/Ne, red diode, and blue diode lasers available for use by researchers throughout the School as well as others inside and outside the University. Three photomultiplier tubes (PMTs) are available for capture of up to three distinct fluorescent emissions simultaneously. With this laser configuration, the microscope is equipped to capture most fluors with excitations ranging from 340 to 650 and emission from 400 to over 700, allowing co-localization of three fluorescent markers in a single image, with the capability of capturing up to six distinct fluorescent signatures per image.
The microscope attached to the confocal is an inverted Nikon microscope equipped with 40X air, 60X oil, and 100X oil objectives and filters to view fluors with emission peaks ranging from 450 to over 700. The scope is also outfitted with Differential Interference Contrast (DIC) optics for transmission images of the tissue or cells as well as 10X, 20X, and 40X phase objectives with an associated long working distance lens and phase rings to capture transmission images by phase microscopy. This combination of objectives, light imaging, and fluorescent filters can serve a variety of functions."
"The software used to run the confocal microscope is Lasersharp 2000, which automates the image collection process, eliminating the need to change filters or add beam splitters depending on the application. The software allows programming for capture of up to 3 fluors at a time over a time course or 3 dimensional space and vertical sectioning. Analysis of the images using Lasersharp 2000 includes 3-D reconstruction and histogram analysis of fluorescent intensity. This software is available in an “off-line” version to any user."
"A very neat and powerful program that reads Bio-Rad PIC images. It allows users to make format conversion, projection, movies, color encoding, montages, etc. Those who use Metamorph (Universal Imaging) will still find Confocal Assistant useful. It is a lot faster than Metamorph when you make projections. Best of all, it's free!
Developed by Todd Clark Brelje"
"MetaMorph Software provides all the tools needed to perform meaningful analysis of acquired images. Common tools for thresholding, filtering, and feature extraction are included as one would expect. However, MetaMorph Software provides other more sophisticated tools such as the integrated morphometry analysis (IMA) module. IMA allows objects to be measured and separated into user-definable classes, based on any combination of measured morphometric parameters, such as shape, size, or optical density. A point-and-click interface, linking images and spreadsheet results, makes it easy to choose representative objects for classification and measurement.
MetaMorph Software also offers many user-friendly application modules for biology-specific analysis such as cell signaling, cell counting, and protein expression. Each application module features a dedicated dialog box with intuitive setting selections, improved segmentation through adaptation to local content, as well as both field and cell-by-cell data logging. After analysis is complete, users can interactively view tabular and image results side by side for their cellular imaging application."