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The inverted microscope series at the center of bioscience's most advanced imaging techniques


Scientists have overcome many live cell imaging challenges using advanced techniques such as TIRF, confocal, FRET, photo activation and microinjection. At the center of all this is the Eclipse Ti, a powerful system that provides instant access to all these methods plus revolutionary Nikon CFI60 optics. Available in three models, the Ti series offers improved system speeds, increased flexibility and efficient multi-mode microscopy as part of a fully-integrated microscope system that is ideal for high-end research and live cell imaging.
 
Stable time-lapse imaging with automatic focus correction system

The Ti-E comes with a unique Perfect Focus System (PFS) that automatically corrects focus drift in real time during a prolonged period of time-lapse imaging.
Focus drift resulting from a temperature drop when reagents are added is instantaneously corrected and the rapid change of cells can be captured.
The incorporation of the PFS in the nosepiece unit saves space and allows two optical component levels to be attached simultaneously utilizing stratum structure.
  EB1 and tubulin in the cortex of Physcomitrella patens moss
Images were acquired on a spinning disk confocal with a Plan Apochromat VC 100x 1.4 NA lens at the Marine Biological Laboratory.
Movie courtesy of: Drs. Jeroen de Keijzer and Marcel Janson, Wageningen University, and Dr. Gohta Goshima, Nagoya University.
 
Concept of the Perfect Focus System

The diagram shows the case when an immersion type objective is used. A dry type objective is also available.
 
Correction to focus drift when reagents are added

  With PFS

  Without PFS
 

Maintaining focus at greater depths

Focus drift can now be corrected in a broader range of Z-axis planes than ever before. Maintaining focus at greater distances from the objective lens and at greater depths within the specimen is also possible.
In addition, PFS's focus drift correction range has been widened, resulting in more reliable and stable data.
     Because PFS can maintain focus at greater depths
   within the specimen, whole images of intersegmental
   vessels sprouting upward from the dorsal aorta are
   clearly captured. Shown in the three channels are three
   different timepoint volumes (red: 0 mins, green: 110
   mins, blue: 240 mins).
   Specimen: vasculature of a zebrafish embryo (95-186 ąým
   away from the coverslip).
   Movie courtesy of: Dr. Robert Fischer, Marine Biological
   Laboratory
 

Improved performance in broader wavelength range


By now employing 870nm wavelength for the coverglass interface detection, near-infrared fluorescence dyes including Cy5.5 can be used. Nikon offers two PFS models, one for UV-visible wavelength imaging and one for multiphoton imaging. The multiphoton model can correct for focus drift even when imaging with wavelengths ranging from 880-1300 nm.
 
Compatible with plastic dishes and well plates

In addition to glass bottom dishes, plastic dishes can be used with PFS. The system is especially suited to high-throughput screening applications that involve multi-well plates.
 
High-speed motorized control and acquisition

The operational speeds of motorized components such as the nosepiece, fluorescence filters and stage have been greatly enhanced, allowing high-speed screening image capture during multi-dimensional experiments. Faster device movement and image acquisition reduce overall light exposure and subsequent photo-toxicity, leading to more meaningful data. The digital Controller Hub significantly increases motorized accessory speed by reducing communication overhead time between components, boosting total operation speed.
   Multipoint snapshots of HeLa cells transiently
   expressing Venus-tubulin and mCherry-actin and
   stained with Hoechst33342 and DiD. (All in pseudo-color)
   Dr. Kenta Saito, Research Institute for Electronic
   Science, Hokkaido University and Dr. Takeharu Nagai,
   The Institute of Scientific and Industrial Research,
   Osaka University
 
High-quality phase contrast images using high NA lenses

The revolutionary external phase contrast unit incorporates a phase ring and allows the use of high NA objective lenses without a phase ring for phase contrast observation. Because there is no light loss due to a phase ring, bright "full intensity" fluorescence images as well as high-resolution phase contrast images can be captured using the same objective lens.
 
Advanced integration with peripherals via intelligent software

Nikon's comprehensive imaging software NIS-Elements provides an integrated control of the microscope, cameras, components and peripherals. The intuitive GUI and efficient workflow make the programming of automated imaging sequences up to 6D (X, Y, Z, time, wavelength, multipoint) easy to perform.
6D time-lapse imaging system

 
Ti-LAPP modular illumination system

The new Ti-LAPP system provides a wide range of illumination modules that can be flexibly combined to create an imaging system tailored for your research. The modularity of the Ti-LAPP system also provides flexibility when the system configuration needs to be changed, an important feature in core imaging facilities and labs that have changing imaging needs.

Ti-LAPP Modular Illumination System


DMD module

The DMD module enables photoactivation and photoconversion of a user-specified pattern and position(s), whereas the conventional FRAP unit only enables photoactivation of a single, manually-positioned spot.

A mouse embryonic fibroblast co-expressing mCherry-tagged lamin A (red) and photo-activatable GFP-tagged lamin A was photo-converted (green) in the lower right region using the DMD module and 405 nm LED light. Time-lapse images were captured using the epi-fluorescence illuminator. By photoactivating a sub-population of the lamin proteins, one can observe their dynamics and subunit-exchange behavior.

Image courtesy of Drs. Takeshi Shimi and Bob Goldman, Northwestern University Medical School

H-TIRF Module

The H-TIRF module automatically adjusts the focus and incident angle of the laser for TIRF observation by monitoring the reflection beam. An even field of TIRF illumination is achieved with a gradation neutral density (ND) filter.


Three-color TIRF image
Using the gradation ND filter, a very even TIRF illumination is achieved. An in vitro preparation of fluorescently-labeled microtubules (tetramethylrhodamine and Alexa 647) and tubulin binding proteins (Alexa 488) was imaged using the H-TIRF illuminator and the gradation ND filter. Incident angles can be automatically adjusted for multiple wavelengths.

Image courtesy of Melissa Hendershott and Dr. Ron Vale, University of California, San Francisco

Flexible module combination

The Ti-Lapp systemí»s modularity and flexible configuration capability provide custom imaging solutions for individual research needs. Modules can also be easily exchanged or added to adapt to changing experimental needs, an important feature for labs with evolving research directions and multi-user, core facilities. For example, by adding a second TIRF module to a single-TIRF configuration, users can easily carry out anisotropy experiments and fast, multi-angle TIRF experiments. Adding a photoactivation/conversion module such as the DMD or FRAP module enables tracking of a sub-fraction of a protein population, providing insights into protein behaviors that would otherwise be illusive when imaging the entire population.




Two-tiered configuration capability

Taking advantage of the Nikon Tií»s stratum structure, modules can be incorporated as two separate layers with multiple modules per layer. Using a dual layer configuration enables optimal filter configuration for each illumination module. This enables optimal filter selection and improves experimental accuracy whilst maintaining the highest acquisition speeds.

  For more information
 
Multiport design with a maximum of five imaging ports

Use of an optional back port enables multiple wavelength FRET imaging with multiple cameras. Moreover, by adding an eyepiece tube base unit with a side port, a maximum of five imaging ports* including left, right and bottom ports are available. (*With Ti-E/B model with bottom port)
Three models for a wide range of applications

Ti-E

The flagship model that is fully motorized for automated multimode image techniques and acquisition
(A model with a bottom port is also available.)
Ti-U

The universal model with the potential for diverse laser illuminators and motorized components
(A model with a bottom port is also available.)
Ti-S

The basic model that can be dedicated to specific tasks