Motorized spinning X,Y TIRF system for even TIRF illumination
Fluorescence microscopy is a balance between light dose, resolution, signal-to-noise, and field of view. Total Internal Reflection Fluorescence (TIRF) microscopy uses an evanescent wave of energy immediately adjacent to the coverslip to excite fluorophores in the specimen. The excitation occurs about 100nm from the coverslip, which is perfect for live cell imaging including focal adhesions, membrane dynamics, receptor function and single-molecule in vitro imaging. TIRF requires placing a narrowly focused beam at the edge of the back focal plane of an objective with NA greater than 1.38 and typically has some shadowing and direction-related artifact in the resulting evanescent wave illumination. Spinning the beam via galvo mirrors around the periphery of the back focal plane at high-speed averages out any artifacts and results in smooth, evenly illuminated images.
Why TIRF?
TIRF microscopy creates an evanescent wave that is about 100nm thick above the coverslip leading to a very thin plane of excitation, even when compared to lightsheet microscopy. This allows scientists to image events on the glass surface with essentially no background when compared to traditional widefield imaging.
Epi Illumination
TIRF
Spinning illumination for even TIRF imaging
VectorTIRF allows for TIRF imaging with high speed and unrivaled clarity on a comparatively large, evenly illuminated field via spinning the excitation light around the back focal plane. Spinning of the excitation light leads to more uniform imaging of thin samples without shadowing and polarization artifacts, as seen in the diagram on the right. VectorTIRF can be used for a variety of live-cell, in vitro, and even single molecule imaging when combined with a modern sCMOS or EMCCD detector. The flexibility of VectorTIRF allows integration with all research inverted microscope frames.
Traditional TIRF
VectorTIRF
Image courtesy of Dr. Emmanuel Derivery, University of Geneva, Dr.Gonzalez-Gaitan lab and Dr. Nicolas Chiaruttini, University of Geneva.
Super Resolution Imaging
Standard VectorTIRF (left) and thunderSTORM reconstruction (right) of Alexa647-labeled microtubles in COS7 cells.
• Up to 200mW lasers
• Fiji direct import of SlideBook .sld files
Images courtesy of Chris de Graffenried at Brown University.
Marianas Multimodal Platform
Combine VectorTIRF with spinning disk confocal, photoactivation, and/or ablation systems for incredible flexibility on one system. Seamless control and switching achieved via SlideBook.
SlideBook Control
- Controls for spinning or fixed point illumination
- Easy control of X/Y centering, spin diameter, and spin duration
- Seamless integration into all SlideBook capture modes (streaming, multiwell, 6D capture)
- Ability to do single diameter, simultaneous multi-color excitation
Images courtesy of Chris Bakal and Oliver Inge at the Institute of Cancer Research in London.
Specifications
LASERSTACK LINES
405nm, 445nm, 488nm, 515nm, 561nm, 594nm, 640nm
SOFTWARE
SlideBook software for image acquisition and analysis
FLEXIBILITY
Polarization input adapter for imaging with two orthogonal polarized input beams
ILLUMINATION FIELD
10 mm with variable beam expansion for fine tuning of excitation power
COMPATIBILITY
Mounts to the sideport on Zeiss, Nikon, Olympus, and Leica inverted microscopes