Research is NOT routine, so customized development is often needed. For new techniques in sample preparation, equipment development, and image analysis, ask us about collaborating with you! New techniques ultimately become new services, so everyone benefits.
As part of our standard consultation, we discuss available strategies to adapt existing equipment for your application, often demonstrating the necessary customization for you. However, more sophisticated customization may be required, both of equipment or software. We have the capabilities for such customization, but the development time is significant. In any case, ask us about how we can help you get a more productive experience at the Facility. You might be surprised that you’re not the only one and that it’s already feasible!
Developing Sample Preparation Protocols
Good fixation is a delicate balance between permeabilization and cross-linking, destruction and stabilization. Some custom protocols that we’ve developed are:
- EM: phalloidin-stabilized actin cytoskeleton of leading edge of motile cells
- High-pressure freezer: use 20atm to prevent ice crystals when freezing, and while ‘vitreous’ ice is maintaining ultrastructure, use fixatives directly on frozen samples!
Customizing Microscopes
Microscopy is always evolving with new sensors, new dyes, and new techniques. Some customization that we’ve developed are:
- Customized optical filter sets for novel fluorophores: multiple labs have already imaged their Hopkins-invented FRET pairs and newly developed fluorophores here at MicFac!
- Special optics for both excitation-ratio and emission-ratio fluorescence.
- Special optics for photoactivation caged molecules (e.g. ATP) and ‘highlighter’ fluorescent proteins (e.g. mEOS, KikGR)
- Customized imaging acquisition protocols
- Dual-viewer for truly simultaneous recording of two-color fluorescence.
- Micro-flow chambers for a variety of applications
- Novel microscopy: developing ‘super-resolution’ fluorescence imaging that approaches EM resolution! SIM, dSTORM, “Hyvolution”, and LLSM are all available.
Customizing Analysis
Microscopy can quickly generate more data than a person can analyze by hand. We have the latest software to automate many tasks, but some tasks require customization and knowledgable analysis. Some customization that we’ve developed are:
- Population counting: e.g. co-transfection efficiency across multiple dishes
- Compare and quantitatively test kinetic models that explain changes in fluorescence (FRAP or FRET) data
- High-content analyses across >5000 samples (an overnight run on the MolDev IXM high-content imager)
- New tools to help interpret co-localization data