Digital Controller Hub significantly increases motorized accessory speed

The newly developed digital Controller Hub significantly increases motorized accessory speed by reducing the communication overhead time between components, boosting total operation speed.

PC control and automation of the Ti’s motorized components are optimized to reduce the respective communication time between action commands and movements producing high-speed total control. By adding firmware intelligence to the microscope, total operation time of the motorized components is reduced. For example, the total time for continuous image acquisition in three modes (two-channel fluorescence and phase contrast) with illumination shutter control is greatly reduced enhancing cell viability.

Remarkably Fast Image Acquisition

Screening image capture of 96 wells in three modes (two-channel fluorescence and phase contrast) is possible at a speed of more than twice that of conventional models.

Nikon’s exclusive and integrated Perfect Focus System (PFS) eliminates focus drift

Focus drift is one of the biggest obstacles in time-lapse observation. Nikon’s PFS design corrects focus drift during long-term observation and when reagents are added. Even with high magnification, high NA objectives and techniques like TIRF, your images are always in sharp focus. Additionally, incorporating PFS in the nosepiece unit saves space and does not limit the use of the Ti expanded infinity space stratum structure.

The PFS employs high-performance optical offset, making real-time correction in the desired Z-plane possible. The state of the PFS is prominently displayed on the front of the microscope. Moreover, when the PFS is not in use, the optical component of the PFS can be simply retracted from the optical path.

Compatible with diverse fluorescence dyes with 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. As the optical characteristics from ultraviolet to infrared range are also improved, the number of usable objectives is increased, realizing stable focus in applications requiring a wide range of wavelengths from Ca2+ concentration measurement in the UV to laser tweezers in the IR.

NIS-Elements Imaging Software provides secure system control

Nikon’s original imaging software NIS-Elements provides an integrated control of the microscope, cameras, components and peripherals and allows the programming of automated imaging sequences. The intuitive GUI makes setting of the experiment parameters easy and reproducible. NIS-Elements offers many tools and controls to facilitate flexible and reliable data acquisition, paired with a diverse suite of analysis tools for measurement, documentation and databasing.

6D/4D Packages Selectable Based on Application

NIS-Elements Ar (advanced research) package allows image acquisition up to 6D (X, Y, Z, time, Lambda (wavelength), multipoint) and analysis or NIS-Elements Br (basic research) which allows up to 4D image acquisition are available depending on research purposes and specimens. Upgrades are also possible by adding diverse optional modules.

For more information on NIS-Elements,

 

High-quality phase contrast imaging

Nikon’s world-leading optical designers have developed the unique “full intensity” external phase contrast unit. With this revolutionary system, a phase ring is incorporated in the microscope body instead of the objective lens, allowing the use of specialized objectives without phase rings and acquisition of high-quality images with high NA objectives. Moreover, using the objectives without a phase ring enables capturing of “full intensity” bright fluorescence images.

Phase Ring Incorporated in the Microscope Body

Incorporating a phase ring—that was normally positioned within the phase contrast objective lens—into the external phase contrast unit optically allows use of specified high NA objectives to produce high-resolution phase contrast images. Four types of phase contrast rings are available according to the objectives used (common for Ti-E/U/S).

Unprecedented High Resolution

Nikon’s high-performance objective lenses, including the 60x and 100x TIRF objectives with the world’s highest numerical aperture of 1.49 incorporating spherical aberration correction collars, deliver high-resolution phase contrast images that can not be captured with any standard phase contrast objective.

Bright Fluorescence Image Using Same Objective

Because there is no light loss due to a phase ring, bright “full intensity” fluorescence, confocal and TIRF images can be captured using the same objective as well as providing phase contrast observation.

Phase Contrast Observation with Water Immersion Objective

It is now possible to use a water immersion objective for phase contrast observation. Clear, high-resolution—refractive index matched—phase contrast images with minimal aberration of deep specimen areas can be captured.

High Resolution Effective for Image Analysis
 

Because phase contrast observation is also possible with the same objective used for TIRF observation as well as DIC observation, phase contrast images with less oblique background shading than that of DIC observation are captured, allowing high-precision data processing and image analysis such as cell contour definition of TIRF image specimen

Multiport and Stratum Structure Support Advanced Research

Multiple image port design with left, right, and bottom* ports for optical output enables a camera or detector to be attached to each port. Furthermore, the expanded space stratum structure enables addition of an optional back port. These features allow image capture with multiple cameras using two-tier dichroic fluorescence filter turrets.

*Available with Ti-E/B and Ti-U/B models with bottom port

Back Port Enables Multi Camera Imaging

Use of an optional back port expands the image capture capability. Used in combination with the side port it allows image acquisition for two wavelengths with two cameras. For example, when observing interaction between fluorescence proteins with FRET (Förster Resonance Energy Transfer) and intensity difference between CFP and YFP is great, individual camera sensitivity adjustment allows comparison of high S/N ratio images.

Stratum Structure Enables Flexible Extendibility

The Ti employs the stratum structure that takes advantage of infinity optics. In addition, the PFS is incorporated in the nosepiece unit, allowing two optical component levels in addition to the PFS to be attached by using the “stage up position set.”

Simultaneous mounting of laser tweezers and photo activation unit as well as multiple stacked epi-fluorescence filter turrets is possible. Each of the tiered motorized filter cube turrets can be controlled individually.

Fluorescence Illumination Functions

The Ti series provides a diverse choice of fluorescence illuminators to support cutting-edge research of cell biology, molecular biology and biophysics using the new imaging and photo activation technologies.

Motorized Laser TIRF for Observation of Cell Membrane Dynamics and Single Molecule

When a specimen is exposed to laser illumination at an incident angle greater than a critical angle, total internal reflection occurs. Under these conditions an evanescent wave is only generated within a couple of hundred nm from the coverslip-specimen interface. By using this light to excite coverslip-specimen interface, fluorescence images with an extremely high S/N ratio can be acquired. This is the principle of TIRF. Nikon’s objective lenses for TIRF observation feature high NA of 1.49, at nearly the theoretical limit for standard oil immersion, and the high S/N technique can capture even single molecule fluorescence images.

The Ti's newly developed motorized laser TIRF illumination unit allows laser incident angle adjustment, shutter control and switching to widefield fluorescence excitation with the control pad or NIS-Elements software. The laser incident angle can be stored with a single touch of the control pad button. Stored laser incident angles can be easily reproduced. This enables alternate time-lapse recording between fluorescence and multi-wavelength TIRF images.