The epi-mounted Vector3 and sideport-mounted Vector2 TIRF are motorized spinning X,Y TIRF systems with homogenous TIRF illumination across a large field-of-view with excellent resolution and signal-to-noise. Vector3 and Vector2 TIRF use galvo mirrors to spin an excitation laser around the periphery of the objective’s back focal plane. This produces an even evanescent wave with very low penetration depth, perfect for imaging live cell focal adhesions, membrane dynamics, and receptor function as well as in vitro experiments that require the thinnest optical section. Motorized focus enables precise TIRF alignment and focus correction across all wavelengths and multiple objects. Illuminate a large, flat field with Vector2 across a broad range of microscopes, or upgrade to the epi-mounted Vector3 for an even larger field and built-in photomanipulation.
Vector3 epi-mounted TIRF and photomanipulation
Vector3 is a motorized spinning X,Y TIRF system with built-in photomanipulation and widefield fluorescence capabilities. Vector3 offers an expansive TIRF field-of-view (FN22) designed for modern sCMOS cameras. Vector3 is capable of galvo-scanned photomanipulation of diffraction-limited spots via user-drawn ROIs for easy FRAP and photoconversion experiments. Incorporation into the epi path of a microscope allows the use of any existing microscope cameras – for example, cameras attached to a CSU-W1 spinning disk – keeping the alignment identical between different imaging modalities and enabling powerful, multimodal imaging experiments. Integrated collimating optics are compatible with most LED illuminators (via a liquid light guide) for widefield illumination.
Vector2 TIRF
Vector2 TIRF is a highly capable and flexible system for spinning TIRF and HILO microscopy supporting most research-grade inverted microscope frames. Optimized for use with 512 x 512 EMCCDs (FN12) for sensitive TIRF imaging of biophysical experiments. The addition of mSwitcher enables combining TIRF and SDC imaging with 10 ms switch times.
Why TIRF?
Fluorescence microscopy is a balance between light dose, resolution, signal-to-noise, and field of view. 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. TIRF microscopy creates an evanescent wave that is about 100nm thick above the coverslip leading to a very thin plane of excitation. 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
Starting with a single point at the edge of the back aperture, Vector3 and Vector2 TIRF spin the beam around the periphery creating an evanescent field from all angles producing shadowless images. Both systems can be used for a variety of live cell, in vitro, and single molecule imaging when combined with a modern sCMOS or EMCCD detector. Vector3 is compatible with Zeiss Axio Observer and Nikon ECLIPSE Ti2 inverted microscope epi-fluorescence ports. Vector2 TIRF allows integration with all research-grade inverted microscope frames.
Super Resolution Imaging
Lasers up to 2W and SlideBook compatibility with analysis pipelines in Fiji (and other software) allows for easy-to-use super-resolution reconstruction.
Standard VectorTIRF image (left) and thunderSTORM reconstruction (right) of Alexa647-labeled microtubules in COS7 cells. Images courtesy of Dr. Chris de Graffenried at Brown University.
HILO Microscopy
Vector3/Vector2 TIRF is also designed for highly inclined and laminated optical sheet (HILO) microscopy. HILO illumination (shown in orange) is achieved just before reaching the critical angle required for TIRF evanescent wave illumination (shown in blue). HILO offers high signal-to-noise with greater penetration for imaging beyond the membrane to the nucleus and through the cell body. Combined with LaserStack and a selection of high and low-powered lasers, Vector3/Vector2 TIRF are versatile single-molecule imaging systems in both TIRF and HILO modalities.
Marianas Multimodal Platform
Combine 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
