All buttons and control switches for motorized operation are designed considering ease of operation, visibility and understandability. Users can concentrate on their research without being hindered by microscope operations.

Operation Buttons on Both Sides and Front of the Microscope Body

Fluorescence filter changeover, objective changeover, objective retraction, Z-axis coarse/fine changeover, PFS on/off control and offset storage, diascopic illumination on/off control can be operated quickly with easy-to identify buttons on the microscope body.

Newly Developed Joystick and Ergonomic Controllers

High-speed motorized XY stage and Z-axis can be controlled using the joystick or ergo controller units. The joystick also allows a custom programmed speed adjustment with precise and natural operational feel.

VFD Screen and Operation Buttons on the Front of the Microscope Body

Microscope status including attached objective information and on/off condition of the PFS can be confirmed on the display at a glance.

PFS Offset Dial

The PFS offset is within easy reach to facilitate control. Coarse/fine switching is possible with simple button operation.

Remote Controller Touch Panel and Preset Buttons

The microscope can be operated and microscope status is confirmed with icons. Also, observation conditions can be memorized with preset buttons. This enables switching observations from phase contrast to fluorescence with a single touch of a button, allowing the user to concentrate on observation without stress or averting attention from the task.

Sophisticated Original Slant Design

By inclining the front part of the microscope’s body slightly backward the distance between the operator’s eyepoint and the specimen has been reduced by about 40mm, improving visibility and ergonomic design.

Nikon’s uncompromising optical technologies provide diverse multi-modal visual information of a specimen using any observation method, delivering the full range of cellular details to researchers.

Nomarski DIC

The perfect balance of high contrast and high resolution is imperative for the observation of smaller structures. Nikon’s unique DIC system is designed to achieve uniform high-resolution images even at low magnifications. The new DIC sliders (dry types) include high-resolution and high-contrast choices.

A filter cube style DIC analyzer can be mounted on the motorized filter turret to minimize switching time between DIC observation and fluorescence observation.

Phase Contrast

For critical phase contrast observation, the CFI Plan Fluor ADH 100x (Oil) objective is available. This objective reduces halos and doubles the contrast of minute cell detail compared to conventional phase contrast objectives. It enables phase contrast observation of specimens with low-contrast minute structures within the cell.

Darkfield

Use of high NA condenser allows darkfield observation. Long-term observation of nanoparticles without photobleaching is possible.

Hoffman Modulation Contrast®

The combination of dedicated HMC objectives and HMC condenser components creates high contrast 3D-like images without halos, of living transparent specimens grown in plastic dishes.

CFI S Plan Fluor ELWD/ELWD Phase Contrast Objectives

Newly developed broadband multilayer coating realizes high transmittance from near-ultraviolet (Ca2+) to near-infrared wavelengths, with improved chromatic correction. The correction collar ring allows these objectives to be used with a diverse range of culture vessels and specimen thicknesses. High-quality images with no aberrations can be obtained under a broad range of illumination techniques.

Plan Apochromat Objective 20x

The new 20x objective is added to the Nikon’s exceptional VC objective series that are effective for digital imaging with complete aberration correction to the very edge of the captured field of view. With this new objective, axial chromatic aberration has been corrected to the violet range (405nm), making it ideal for confocal observation and photo activation.

The synchronized control of many motorized components such as the nosepiece, fluorescence filters, shutters, condenser turret and stage, allows researchers to use the microscope for a wide range of automated multi-dimensional experiments. Faster device movement and image acquisition decrease overall light exposure and subsequent photo-toxicity, leading to more meaningful data.

Operation and/or changeover speed of objectives, filter cubes, XY stage, excitation/barrier filters has been greatly enhanced, realizing stress-free operational environment that enables researchers to focus on observations and image capture routines. The newly developed controller that memorizes and reproduces observation conditions and the joystick that enables stage control at will make the microscope feel like an extension of your eyes and hands.

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.

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.

Focus drift is one of the biggest obstacles in time-lapse observation. Nikon’s Perfect Focus 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.

For more information on PFS, click here.

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.

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, click here.

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.

Photo Activation for PA-GFP Observation

When fluorescence proteins such as Kaede and PA-GFP are exposed to 405nm illumination, fluorescence characteristics change and fluorescence emission colors shift. These proteins are used for selectively marking proteins of interest within cells and studying their dynamic interactions. The Ti series features a specialized photo activation illuminator that allows fluorescent time-lapse observation of dynamic events following photo activation or photo conversion.

FRET for Analysis of Intracellular Ca2+ Concentration

Using FRET (Förster Resonance Energy Transfer) technique, intermolecular interactions between molecules within close proximity of one another can be detected and measured. Using the optional back port, each FRET channel can be separated by wavelength and sent to separate cameras. This enables the capture of high-resolution images in the entire frame for each wavelength. Even when intensity difference between wavelengths is large, a high-quality FRET image can be captured by adjusting camera sensitivity for each wavelength.

White Light TIRF Utilizing Mercury or Arc Lamp Illumination

Mercury arc lamp illumination can be used for TIRF observation. The specialized high-performance epi-fl illuminator unit (white light TIRF) allows multi-spectral TIRF to be accomplished without multiple lasers. The wide wavelength band of mercury illumination makes multiple wavelength TIRF observation possible by simply changing filter cubes.

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 Extendability

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.

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.