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Services and Equipment
Optical Imaging CoreTraining
The Optical Imaging Core offers training in a wide range of methodologies for quantitative imaging. These include but are not limited to:
- Fluorescence-based histology and fluorescent analysis of gene expression
- Multiphoton confocal microscopy
- Live-cell imaging
- Cell motility assays
- Cell signaling and changes in cell state
- Visualizing 3-D objects through optical sectioning
- Spectral deconvolution of complex fluorescent signals
- Fluorescence recovery after photobleaching (FRAP) and related techniques (e.g., FLIP)
- Photoactivation and photoconversion
- Colocalization
- Structural analysis of organelles
- Quantitative tracking of motile organelles and subcellular particles
- Membrane trafficking assays
- Assays for protein-protein interaction
- Förster resonant energy transfer (FRET)
Equipment
Nikon A1plus-RSi scanning confocal microscope
- 403, 488, 561 and 640 nm solid-state lasers
- SOLA solid-state LED epi-illuminator
- Two alternative confocal detection modes:
- 4-detector “DU4” setting with high-sensitivity GaAsP detectors in the red and far-red detection channels
- 32-detector spectral imaging mode
- Transmitted light detector in confocal mode
- Two scanning modes (Galvo scanner and high-speed resonant scanner)
- DualView2 beam splitter with Hamamatsu Orca Flash2.8 sCMOS camera for high-speed quantitative FRET imaging
Training in imaging data analysis is available. Depending on the complexity, this can be done either within the Optical Imaging Core or in consultation with experts in VARI’s Bioinformatics and Biostatistics Core. Protocols include:
- Methods for converting image data to numeric values
- Statistical best practices
- Freeware for image analysis (ImageJ) and the quantitative handling of very high-throughput data sets (CellProfiler, Endrov)
- Software for deconvolution and 3-D analysis (Huygens Suite)
Regardless of whether the protocol of interest is listed above, the facility is eager to work with scientists to develop rigorous and quantitative assays to address specific research needs.