IMC services include training and access to digital fluorescence microscopy and transmission electron microscopy.
Standard and High Resolution Microscopy
For standard fluorescence microscopy, a Zeiss AxioPlan II upright microscope is available, equipped with PC-controlled, high-resolution black-and-white and color CCD cameras. Brightfield capabilities include standard brightfield, phase, DIC and dark field illumination, with a range of magnifications from 2.5x to 100x using high quality objectives.
High-resolution fluorescence of thin samples (cell monolayers or sections less than 20 microns) is available via Applied Precision's DeltaVision Restoration Microscopy system, a deconvolution-based, wide-field optical microscope. This instrument is fully automated for Z-series acquisition, with up to four fluorescent channels. Available filter sets include the standard Sadat set (DAPI, FITC, Rhodamine, Texas Red, Cy-5), and a live cell filter set with CFP, GFP, YFP and RFP. Image data collection and interpretation is made with SoftWorx image analysis and model-building software. A stand alone SoftWoRx workstation is also available for further image analysis.
Laser or Digital System
Confocal microscopy is available using the Zeiss LSM 510 META confocal laser scanning microscope with a triple laser system and sophisticated AOTF laser control that can eliminate channel bleed through. CFP, GFP, FITC, Rhodamine, Texas Red and Cy-5 can be imaged, along with DIC. The META upgrade is a versatile spectral imaging detector that allows spectral unmixing and at least 4 channel acquisition. Bioptechs environment chambers are also available for pre-approved live cell imaging projects, such as time lapse and photobleach/recovery studies.
Transmission Electron Microscopy
The TEM facility is equipped with a digital Hitachi H-7500 Electron Microscope, a 2Kx2K Gatan CCD, and excellent, web-enabled database/software capabilities. Available EM services include standard fixation, embedding, ultramicrotomy and stains. Special stains and immunogold labeling are also available on a limited basis.
We provide researchers assistance with all aspects of assay designing and development. Each assay is custom designed for optimal results, from experimental design and image acquisition to data analysis and presentation using Pipeline Pilot for image analysis. This resource provides consultation and technical support for pilot experiments using coverslips, 96- or 384-well plate formats in single cell fluorescence-based imaging studies and quantitative image analyses.
Potential assays include nuclear translocation, reporter gene analysis, cell cycle, apoptosis, and RNAi gene silencing analysis, to name some of the more common assays performed in the IMC. To take advantage of this service, principle investigators are required to submit a one page proposal describing the goals of the assay to Dr. Michael Mancini via e-mail. An ad hoc panel will assess proposals for compatibility/feasibility with the current resources.
Access, Tutorials and Fees
A hierarchal access structure is in place for investigators, depending upon affiliation with supporting grants, or with the Department of Molecular and Cellular Biology. An Investigator/Project Profile form is required for everyone in order to determine the most appropriate instrumentation/service and access privileges.
Tutorials are required for all core users on all instruments. Fees vary per microscope from $20-50/hour; fees for imaging workstations are $10/hour; labor charges are also at $40-80/hour. Core-approved users can arrange for independent use during peak time (8:30 a.m.-6 p.m.), or, with significant training, off peak time at discounted rates (evenings, weekends).