Inverted Research Microscope ECLIPSE Ti

Inverted Research Microscope ECLIPSE Ti
Inverted Research Microscope ECLIPSE Ti
● Advanced functions of Ti-E dramatically expand research imaging possibilities
At the Center of Your Research Discoveries
The Essence of Cutting-edge Microscopy Research
Fast and Automated
High-speed motorized components allow fast, coordinated and seamless image acquisition【P4】
Screening
Multimode scanning of well plate at an unprecedented speed【P5】
Microscopes are critical tools for cutting-edge research in biology, medical and pharmaceutical sciences. To satisfy the demands
of today’s high-end research, Nikon has developed the new Ti series of microscopes. Combined with NIS-Elements imaging software,
the Ti supports diverse image acquisition and analysis methods such as multi-dimensional time-lapse imaging to acquire temporal,
spatial and spectral information of fast, dynamic live cell processes. Intelligently designed automation and further expansion
of Nikon’s powerful modular approach make the Ti ideal for applications such as confocal, FRET, High Content Analysis (HCS),
and photobleaching/photo activation to study interaction of fluorescence protein molecules in living cells and tissues.
Nikon’s exclusive Perfect Focus System (PFS) is now incorporated into the nosepiece unit and allows for the simultaneous use
of two separate levels for additional illuminators or detectors. The newly developed “full intensity” phase contrast unit enables
acquisition of incredible phase contrast images without the use of light-attenuating phase contrast objectives.
Time-lapse Imaging
Built-in Perfect Focus System (PFS) for automatic focus correction【P6】
High-quality Phase Contrast Observation
Newly developed “full intensity” optical components enable phase contrast
with high NA non-phase-contrast objectives【P8】
Multiple Cameras
Image acquisition and analysis with multiple side ports and back port cameras【P9】
Motorized Laser TIRF (Total Internal Reflection Fluorescence) Observation
Alternate time-lapse observation between widefield fluorescence and TIRF (NA 1.49) images by
fast illumination switching and motorized control of laser incident angle【P10】
Photo Activation
The photo activation unit allows cell marking and dynamic analysis using photoactivatable
and photoswitchable proteins such as PA-GFP and Kaede【P11】
Confocal Imaging
Seamless integration with confocal microscope systems for high-performance spectral confocal imaging【P19】
The basic model that can be dedicated to
specific tasks, built with two output imaging ports
The flagship model that is fully motorized for
automated multimode image techniques and acquisition
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The universal model that comes standard with four output ports
and potential for motorized components
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Ti: Stress-Free Operation
Remarkably Fast Image Acquisition!
High-speed Motorized Control
and 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.
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.
Enhanced speed of individual motorized components
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.
● High-speed XY stage movement
● High-speed Piezo Z stage movement
● High-speed epi-fl filter changeover
Nikon-exclusive high-speed encoded stage
Nikon-specified Piezo Z specimen stage
Nikon filter dichroic cube turret
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.
● Control process
■ Conventional model
Signal
communication
Stage movement
Signal
communication
PFS
correction
Signal
communication
Filter changeover
Signal
communication
Image capture
■ Ti-E
Signal
communication
Stage
movement
PFS
correction
Filter
changeover
Image
capture*
Multipoint snapshots of HeLa cells transiently expressing Venus-tubulin and
mCherry-actin and stained with Hoechst33342 and DiD. (All in pseudo-color)
*When used in conjunction with Nikon’s DS-Qi1 or Andor iXon, the experimental protocol is loaded and run from internal memory, eliminating communication overhead between the PC
and microscope and significantly reducing acquisition time. No critical events are lost due to time delays.
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Photos courtesy of: Kenta Saito and Takeharu Nagai,
Research Institute for Electronic Science, Hokkaido University
Ti: Reliable Time-Lapse Observation
PFS and NIS-Elements Realize Stable
and Reliable Imaging
Nikon’s exclusive and integrated Perfect Focus
System (PFS) eliminates focus drift
Comprehensive Imaging Software NIS-Elements
Provides Secure System Control
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 (see page 9).
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.
● Microscope setting
● Time-lapse (camera) setting
● Z setting
● XY (stage) setting
● λ(fluorescence turret) setting
Real-time focus correction
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● Concept of the Perfect Focus System
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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.
● Correction to focus drift when reagents are added
■ With PFS
Specimen
Interface
Coverslip
Oil, water
Objective
Perfect Focus Nosepiece
Near-IR light
LED
Line-CCD
Intuitive GUI and efficient workflow of NIS-Elements make 6D (X, Y, Z, t (time), Lambda (wavelength), multipoint) image acquisition that requires complex
settings easy to perform. Simply by choosing the necessary parameters for each dimension, images are automatically captured and a multi-dimensional
ND2 file is generated, which can be seamlessly viewed, analyzed, and exported. Converting the format of the captured multi-dimensional image
to standard formats is also easy to accomplish.
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Adding reagent
■ Without PFS
Offset lens
Observation
light path
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Adding reagent
Compatible with diverse fluorescence dyes
with improved performance in broader wavelength range
NG108 cell
NEW
Zebrafish larva transgenically expressing lens
specific GFP and stained with Hoechst33342,
acetylated tubulin-Alexa555 and phalloidin-Alexa647
● NIS-Elements 6D time-lapse imaging system
NIS-Elements
software
Camera
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Transmission (T%)
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Photo courtesy of: Ippei Kotera, Shinya Hosaka and
Takeharu Nagai, Research Institute for Electronic Science,
Hokkaido University
Shutter
By combining the Nikon motorized stage, motorized filter turret and “smart”
specified shutters, acquisition of multipoint, multi-channel time-lapse images and
Z-axis information of each of these points is possible.
Motorized XY stage
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Live imaging of primary rat cortical neurons
stained with Hoechst33342 and DiR
Photos courtesy of: Satoe Ebihara, Kaoru Katoh,
The National Institute of Advanced Industrial Science and Technology (AIST)
Photo courtesy of: Kazuki Horikawa and Takeharu Nagai,
Research Institute for Electronic Science, Hokkaido University
By now employing 870nm wavelength for the coverglass interface detection, near-infrared
New PFS
Conventional PFS
fluorescence dyes including Cy5.5 can be used. As the optical characteristics from ultraviolet
Note: Cases without IR-cut filter
to infrared range are also improved, the number of usable objectives is increased, realizing
870nm
stable focus in applications requiring a wide range of wavelengths from Ca2+ concentration 100
measurement in the UV to laser tweezers in the IR.
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The diagram shows the case when an immersion type objective is used.
A dry type objective is also available.
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Camera
3
3
Motorized inverted microscope
Ti-E epi-fluorescence set
Data analysis
equipment (PC)
Joystick unit
400
600
800
1000
1200
Wavelength (nm)
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Ti: Revolutionary Phase Contrast System
Ti: Maximum Flexibility & Expandability
High-quality Phase Contrast Images
with High NA Lens, as well as
Bright Fluorescence Images
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 simultaneous
image capture with multiple cameras using two-tier dichroic fluorescence filter turrets.
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.
*Available with Ti-E/B and Ti-U/B models with bottom port
Phase ring is incorporated in the microscope body
Back port enables multiple camera imaging
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)
Use of an optional back port expands the image capture capability. Used in combination
with the side port it allows simultaneous 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.
Conventional position
of phase ring
New position of
phase ring
ECFP image from YC3.60
cp173Venus image from YC3.60
Photos courtesy of: Kenta Saito and Takeharu Nagai,
Research Institute for Electronic Science, Hokkaido University
Changing the conventional concept of phase contrast
● 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.
● 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.
Back port can be attached as an option.
● 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.
Stratum structure enables flexible extendibility
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C. elegans: Touch neurons stained with EGFP
Photos courtesy of: Motomichi Doi and Kaoru Katoh,
The National Institute of Advanced Industrial Science and Technology (AIST)
NG108 cell: Growth cone stained with EGFP-fascin
Photos courtesy of: Satoe Ebihara, Kaoru Katoh,
The National Institute of Advanced Industrial Science and Technology (AIST)
● Use of laser tweezers without changing lens
Because an objective without a phase ring can be used for phase contrast observation,
use of laser tweezers is possible without changing the objective lens.
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● 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.
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 epifluorescence filter turrets is possible. Each of the tiered
motorized filter cube turrets can be controlled individually.
Example: In addition to the PFS, a photo activation module (upper tier) and a back port (lower tier) are mounted.
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Ti: Advanced Applications
Advanced Fluorescence Illumination Functions Respond
to Leading Bio-imaging from Live Cell to Single Molecule
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.
● Time-lapse imaging by switching TIRF and epi-fluorescence observation
Laser TIRF (Motorized/Manual)
■ TIRF
For observation of cell membrane dynamics and single molecules
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■ Epi-fl
Motorized TIRF illumination unit
NG108 cell: Growth cone stained with EGFP-fascin
Photos courtesy of: Satoe Ebihara, Kaoru Katoh,
The National Institute of Advanced Industrial Science and Technology (AIST)
This unit allows total internal reflection fluorescence observation of
specimens such as cell focal adhesions or single molecules in-vitro
using laser illumination. When used with a high-sensitivity camera,
images with extraordinarily high S/N ratios that allow observation of
single molecules can be captured.
The newly developed motorized laser TIRF illumination unit allows laser
incident angle adjustment, shutter control and switchover to widefield
fluorescence excitation using the control pad or NIS-Elements software.
Laser incident angles can be saved with a single touch of the control
pad button and can be easily retrieved, enabling alternate time-lapse
recording between fluorescence and multi-wavelength TIRF images.
Photo activation
For observation of photo-activated and photo-convertible
fluorescent protein
Photo activation illuminator unit
Remote controller
● Principle of TIRF (Total Internal Reflection Fluorescence)
Low refractive index
(solution)
TIRF objectives feature a high NA of 1.49—very close
to the theoretical limit for standard oil immersion—
and can capture even single-molecule images.
405nm laser spot
High refraction index
(coverslip)
Reflected light
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Evanescent wave at the coverslipspecimen interface, typically within
a couple of hundred nm
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When light is incident to the coverslip at an
angle greater than the critical angle (θ) for
Total Internal Reflection, the light no longer
propagates through the specimen, but sets up
an evanescent field at the coverslip/specimen
interface that can excite fluorescence in the
specimen in an optical section less than
100nm. By exciting such a thin section within
the specimen, extremely high S/N data can be
acquired.
When fluorescence proteins such as Kaede and PA-GFP are exposed to 405nm illumination, fluorescence characteristics change. For example, Kaede
changes fluorescence colors from green to red, and PA-GFP increases fluorescence intensity 100 times. Kaede and PA-GFP are used, respectively, for
selectively highlighting cells and proteins of interest within live specimens and studying their dynamics. The photo activation illuminator utilizes lasers
ranging from 405nm to 647nm to produce target spots of varying diameters, allowing time-lapse observation of dynamic events in living cells.
Incident light
Range of incident angles
greater than the critical angle
Overview of TIRF
CFI Apochromat TIRF 60x Oil (left)
CFI Apochromat TIRF 100x Oil (right)
Photo activation of PA-GFP in a living mammalian cell by 405nm laser irradiation
Photos courtesy of: Tomoki Matsuda and Takeharu Nagai, Research Institute for Electronic Science, Hokkaido University
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TIRF-photo activation
Epi-fluorescence
With the integration of the laser TIRF illuminator and photo-activation unit, both functions are now combined on one microscope.
The user can switch between the two functions with ease.
Chromatic aberration has been significantly improved over a broad wavelength range
to provide sharper and brighter fluorescence images.
Epi-fl illuminator unit
TIRF-PAU illuminator unit
White light TIRF
This unit allows high-performance yet cost-effective total internal reflection fluorescence microscopy
as well as oblique and standard widefield fluorescence techniques using mercury illumination.
The wide wavelength band of mercury illumination makes multiple wavelength TIRF observation
possible by simply changing filter cubes.
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 simultaneously. 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.
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Epi-fl illuminator unit with white light TIRF
5.0
Imaging histamine-evoked Ca2+ release in mammalian cells reported by
a FRET-based Ca2+ indicator, YC3.60
YFP
CFP
Photos courtesy of: Kenta Saito and Takeharu Nagai,
Research Institute for Electronic Science, Hokkaido University
1.0
Photo courtesy of: Yasushi Okada,
Cell Biology, Graduate School Medical
Department, The University of Tokyo
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Photo courtesy of: Richard Cheney Ph.D.,
UNC Chapel Hill
0
20
40
60
80
100
120
Time (sec)
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Ti: Excellent Imaging
Ti: High Performance with User-Friendly Operation
Use of Optimal Optical Technology
for Each Observation Method Allows
Uncompromised Image Capture
Enhanced Operability Enables
Comfortable Observation
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.
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.
Nikon Advanced Modulation Contrast
Fast and comfortable operation with motorized components
Nikon has developed dedicated objectives for advanced modulation contrast. Colorless
and transparent samples can be observed in high relief with a plastic dish, which is not
possible in DIC observation. The direction of contrast can be matched to S Plan Fluor
ELWD NAMC objectives, thereby allowing optimal contrast selection for techniques like
microinjection and ICSI.
● Operation buttons on both sides
of microscope body
● VFD screen and operation buttons
on front of microscope body
● Remote controller touch panel
and preset buttons
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-toidentify buttons on 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.
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.
Photos courtesy of: Gianpiero D. Palermo, M.D., Ph.D., Cornell University
Visual conformation of the buttons can be clearly viewed
in the dark
Nomarski DIC
Darkfield
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.
● PFS offset dial
Use of high NA condenser allows darkfield observation. Long-term
observation of nanoparticles without photobleaching is possible.
● Newly developed joystick and
ergonomic controllers
● Motorized analyzer cube
A filter cube style DIC analyzer can be
mounted on the motorized filter turret to
minimize switching time between DIC
observation and fluorescence observation.
Photo courtesy of:
Jan Liphardt, University
of California Berkeley
Filter cube style DIC analyzer
High-speed position changing of the filter cubes in 0.25 second
The PFS offset is within easy reach to facilitate
control. Coarse/fine switching is possible with
simple button operation.
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.
Highly parallel single-molecule DNA bending assay using darkfield microscopy. Each bright green
spot is a single plasmon ruler, composed of a pair of DNA-linked gold nanoparticles. Enzymatic DNA
bending or cleavage can be monitored by following the intensity and color of the plasmon rulers. For
more information see Reinhard et al, PNAS (2007).
Remote controller
PFS offset dial
Sophisticated original
slant design
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.
Joystick unit
40mm
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.
Ergonomic controller
CFI Plan Fluor ADH
100x (Oil) objective
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Viewed with an ADH objective
Viewed with a conventional
phase contrast objective
Joystick and ergonomic controllers can not be used simultaneously;
they are offered to provide a personal choice of control.
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Motorized Elements for Comfortable Observation
Fast, automatic operation by integrated control
with NIS-Elements software
Ti-E can be fully motorized with the HUB-A
Microscopes have evolved from merely observation devices to software-controlled data acquisition devices.
Nikon’s Ti series not only features fast and comfortable motorized operation, but it also realizes acquisition of
reliable data by controlling all motorized components for automatic imaging with the NIS-Elements imaging software.
HUB-A
Motorized control of laser incident angle and repositioning
by memory settings are possible.
Four components of Ti-U/S can be motorized with the HUB-B
● Nikon motorized XY stage
High-speed shutter for fluorescence excitation and brightfield
illumination (Manufactured by Sutter Instrument Company)
HUB-B
High-speed, precise Z-axis control is possible.
(Manufactured by Mad City Labs, Inc.)
● Motorized nosepiece
● Motorized filter rotating turret
● Motorized shutter “Smart shutter”
By attaching HUB-B unit to the Ti-U/S,
two optional motorized components, such as
fluorescence filter turret and condenser turret,
in addition to the stage and nosepiece, can
be motorized, greatly enhancing flexibility.
● Piezo Z stage
Fast and precise positioning is possible. Suitable for multipoint timelapse observation. (Available as encoded or non-encoded versions)
● Motorized laser TIRF illumination unit
Communication speed is dramatically
increased through proprietary motorization
algorithms, innovatively accelerating the
sequence of operation. The Ti-E assures more
reliable and efficient data acquisition in the
research field.
● Motorized HG precentered fiber
illuminator “Intensilight”
Controls shutter on/off and intensity of fluorescence excitation light.
● Motorized excitation filter wheel
Six objective positions can be changed.
(Photo shows motorized PFS nosepiece)
Position of fluorescence filter cubes can be changed in
0.3 sec. per position. (Photo shows high-performance type)
● Motorized condenser turret
Fluorescence excitation filters (8 positions — using 25mm filters)
can be changed at a high speed of 0.15 sec. per position.
Motorized condenser changeover is possible.
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● Motorized barrier filter wheel
● Remote controller
● PFS offset dial
● Joystick unit
● Ergonomic controller
Fluorescence barrier filter positions (8 positions—using 25mm
filters) can be changed at a high speed of 0.15 sec. per position.
Microscope status can be confirmed with icons.
The microscope can be operated via the touch panel.
Real-time offset amount of Z-axis depth can be controlled
after PFS setting.
Flexible positioning of the motorized stage is possible.
Multiple operations are possible with manual controller.
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Compact, High-Performance CCD Cameras
Advanced Confocal Laser Scanning Microscopes
Digital Sight series digital cameras
for microscopes
Advanced confocal laser microscopes
optimally match the Ti-E
These camera systems allow for smooth integration with a
microscope and other products. Different combinations of
camera head and control unit meet the requirements for any
microscopic image acquisition.
Confocal microscope
Camera heads
■ DS-Qi1
Definitive camera for fluorescence time-lapse
imaging features high sensitivity, low noise, superior
quantitative linear response and quantum efficiency,
wide dynamic range and high frame rate.
■ DS-Fi1c
High-definition 5.0-megapixel cooled color camera
head. Cooling mechanism retains CCD at room
temperature minus 20°C and realizes low noise.
True spectral imaging confocal microscope
● A1R/A1
● A1Rsi/A1si
The A1R with a revolutionary hybrid scanner realizes ultrafast
and high-resolution imaging
High-performance spectral detector supports simultaneous
excitation of multiple wavelengths
• Hybrid scanner capable of high-speed imaging at 420 fps (512 x 32 pixels)
allows simultaneous imaging and photo activation (A1R)
• High-resolution imaging up to 4096 x 4096 pixels
• With the VAAS pinhole unit, flare can be eliminated and image brightness
retained; different sectioning can be simulated after image acquisition
• Dichroic mirror with 30% increased fluorescence efficiency provides high
image quality
• Acquisition of 32 channels (512 x 32 pixels) at 24 fps in a single scan
• Accurate, real-time spectral unmixing
• Simultaneous excitation of four lasers
• V-filtering function adjusts individual sensitivity of up to four spectral ranges,
allowing production of customized filters that are optimal for various
fluorescence probes
Multiphoton confocal microscope
● A1R-MP
High-speed imaging of deep area in a living specimen
NEW
■ DS-Vi1
High-speed 2.0-megapixel color camera head displays
smooth, high-quality live images.
■ DS-Fi1
High-definition 5.0-megapixel color camera head
features high frame rate, high red sensitivity, high
resolution and accurate color reproduction.
• Resonant scanner enables imaging up to 420 fps (512 x 32 pixels)
• Deep imaging with high-sensitivity NDD (non-descanned detector)
• Sharper, brighter imaging with high NA objectives deposited with Nano
Crystal Coat
• High-speed, high-precision unmixing with NDD
• Multiphoton laser beam can be automatically aligned with a single click
A1Rsi/A1si
■ DS-Ri1
Ultrahigh-resolution 12.7-megapixel, 2200TV line
cooled color camera that provides faithful reproduction
of specimen color and fast display of live images.
A Peltier cooling mechanism reduces heat noise.
High-speed imaging during photo activation
While imaging a HeLa cell expressing Kaede with green and red fluorescence using 488nm and 561nm lasers as excitation lights, Kaede in a ROI is continuously
activated with the 405nm laser for photo conversion. The dispersion of Kaede red fluorescence produced by photo conversion can be observed.
Comprehensive Imaging & Analysis Software
Imaging software NIS-Elements
6D/4D packages selectable depending on purpose
Activation laser wavelength: 405nm, Imaging laser wavelength: 488nm/561nm, Image size: 512 x 512 pixels, 1 fps
Photos courtesy of: Dr. Tomoki Matsuda and Prof. Takeharu Nagai, Research Institute for Electronic Science, Hokkaido University
Confocal microscope
True spectral imaging confocal microscope
● C1 plus
● C1si
Personal confocal microscope now supports FRAP
Spectra across a wide 320nm range captured with a single scan
• 1000x optical zoom of ROI
• ROI scanning is possible with an optional AOM/AOTF
• Accommodates a greater variety of lasers with wavelengths ranging from
405 to 640nm
• 4-channel simultaneous acquisition such as 3-channel confocal plus DIC
• High-speed, low-invasive imaging by a single scan acquisition
• Unmixing of spectral images without crosstalk
• Nikon’s proprietary DEES and DISP technology for bright images
• Accuracy of spectra is maintained with diverse correction technologies
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NIS-Elements has been developed by Nikon, a leader in microscope and camera technology.
It allows automated operations from advanced image acquisition to analysis and measurement
by integrating control of microscope, camera and peripherals. It is Nikon’s modular imaging
software ideally integrated for all microscopy applications.
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■ DS-L2
Standalone control unit with high-resolution large 8.4-in.
LCD monitor allows image capture without a PC.
Pre-programmed imaging modes realize optimal imaging
settings by choosing icons of the illumination method.
Annotation, calibration and measurement tools are
provided. Various digital interfaces and networking
function enable images to be shared. Various USB 2.0
media storage, HUB and host control are provided.
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■ DS-U2
USB2.0 PC-use control unit is suitable for operations
from advanced image capture to image processing and
analysis by integrating control of camera, peripherals
and microscope with NIS-Elements imaging software.
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Control units
DS-Qi1
Ti-E
Ar (advanced research) package that allows image acquisition up to 6D (X, Y, Z, time, Lambda
(wavelength), multipoint) and analysis and Br (basic research) package that allows up to 4D
image acquisition are available depending on research purposes and specimens. Upgrades are
also possible by adding diverse optional modules.
NIS-Elements D, designed for easy image acquisition yet powerful and economical, is also available.
18
HeLa cell in which nucleus is labeled with CFP, actin-related protein (Fascin) labeled with GFP, Golgi body labeled
with YFP, and mitochondria labeled with DsRed. Spectral image captured with 408nm and 488nm laser exposure (left).
The fluorescence spectra of the captured image are unmixed using reference spectra (right).
Photos courtesy of: Kaoru Katoh and Ayako Kojima, Neuroscience Research Institute,
The National Institute of Advanced Industrial Science and Technology (AIST)
Ar package
19
Accessories
● Incubator
● Thermal plate warmer ThermoPlate
The internal temperature of the case is maintained at 37ºC. However,
temperature adjustment from room temperature to 50ºC is possible.
The incubator is compatible with both the rectangular stage and the
motorized stage. Various dishes can be used, including a well plate, with
different inside attachments.
Ergonomic Eyepiece Tube
Binocular Eyepiece Tube D
Binocular Eyepiece Tube S
Eyepiece Tube Base Unit/Phase Contrast
Eyepiece inclination is adjustable from 15°
to 45°. Includes darkslide shutter and
Bertrand lens.
Observation of conoscope image with incorporated
Bertrand lens (phase telescope) is possible and
darkslide shutter is provided.
Standard model
High-resolution imaging with “full intensity”
external phase contrast system is possible.
TV port is incorporated.
Eyepiece Tube Base Unit/Side Port
Plain Eyepiece Tube Base Unit
Eye Level Riser
Stage Up Position Set
TV port is incorporated.
Standard model
Two 25mm emission filters can be installed.
Stage height can be raised by 70mm to
mount multiple components utilizing
expanded stratum structure.
Stage Base
Back Port Unit
High NA Condenser (Oil/Dry)
CLWD Condenser
MATS series
A temperature controllable stage ring with a glass heating
plate keeps the specimen at a set temperature.
Temperature is adjustable from room temperature to 50ºC
in 0.1ºC increments.
Manufactured by Tokai Hit Co., Ltd.
For motorized stage
● Stage incubation system INU series
● NT-88-V3 micromanipulator system
It sustains the internal temperature at 37ºC with humidity of 90% and CO2 of
5% to keep the specimen in a stable and precise condition for about three
days. A special technique is employed to minimize focus drift caused by
thermal expansion of a stage. The glass heater on top of the chamber
prevents condensation and enables clear images.
A packaged set of compact instrumentation—about half the size of
a conventional model—for cellular micromanipulation, the NT-88-V3 is ideal
for IVF (in-vitro fertilization), ICSI (intracytoplasmic sperm injection),
electrophysiology, or transgenic biotechnology applications. Hanging joystick
design provides superior ergonomics and operability. Remote oil hydraulic
operation minimizes pipette vibration. An index of the coarse manipulator
enables easy position adjustment of the pipette.
Manufactured by Tokai Hit Co., Ltd.
Manufactured by Narishige Co., Ltd.
20
Stage base for configuration without diascopic Combined use with stage up riser allows a
illumination
camera to be mounted on a back port.
Perfect for observation with high NA objectives For high NA long working distance objectives
NAMC Condenser
Stage Ring
Epi-fluorescence Attachments
Double Lamphouse Adapter
For observation of Nikon Advanced
Modulation Contrast
Acrylic ring (left) features superior objective
lens visibility and the glass ring (right)
features less thermal expansion— ideal for
time-lapse observation.
Light source and illumination optics for high
S/N images
For attaching two light sources
21
System Diagram
D
Eyepieces/Tube Base Units
C L
Illumination Pillars
GM
Nosepieces
Epi-fl Illuminators
λ
H?N?L
A
CFI UW Eyeguard
TI-DIC Lambda Plate
T-P2
P
B
C
Eyepieces CFI 10x, 12.5x, 15x
D
C-CT Centering Scope
T-P2 DIC Polarizer
I
C-LHG1 HG Lamp 100W
TI-PS100W Power
Supply 100-240V
TI-BPU Back Port Unit
TI-C-P
NAMC Polarizer
OPEN
D-LH/LC Precentered
Lamphouse with LC
E J
OFFSET
INTERLOCK
PFS
PIEZO
TE-PS30W/PSE30W
Power Supply A 115V/230V
TI-TD
Binocular Tube D
TI-TERG
Ergonomic Tube
TI-TS
Binocular Tube S
TI-DS Diascopic
Illumination Pillar
30W
∞Ph1
TI-DH Diascopic
Illumination Pillar
100W
TE-C ELWD-S
Condenser
TI-FL Epi-fl
Illuminator Unit
C
TI-TIRF
Stage-Up Lens*5
TI-A DIC
Analyzer Block
Epi-fl Filter Blocks
HMX Lamphouse
TI-C System
TI-CT-E Motorized
TI-C System
TI-CT-E Motorized
Condenser Turret Condenser Turret*1 Condenser Turret Condenser Turret*1
HMC 0.4
ELWD 0.3
JAPAN
YM-EPI 3-3 Pin
Extension Cable
Reflection Mirror*7,
TIRF 60/40 Half Mirror
TI-FLC Epi-fl
Filter Turret
JAPAN
JAPAN
T-C High
T-C High
TI-C NAMC Lens
NA Lens (Dry) NA Lens (Oil)
A
TI-PS100W Power Supply
100W 100V/115V/230V
HQ Filter
Blocks
E J
TI-IER3H Eye Level Riser
Halogen Lamp
Socket 100W
G
MC-TMD2 High NA
Condenser Slider
G
Halogen Lamp
12V100W
TI-SFL Epi-fl Illuminator Unit with
White Light TIRF*3
TE-C ELWD-S
Condenser
TI-T-BPH Eyepiece
TI-T-BS Eyepiece
TI-T-B Eyepiece
Tube Base Unit for PH*2 Tube Base Unit with Side Port Tube Base Unit
C-Mount Camera
Xenon Lamphouse C-XES Xenon
Lamp Socket 75W
HMX4
TE-AT Double
Lamphouse Adapter
HQ Filter Blocks
T-C High NA
Condenser Lens Unit
D
Xenon Power Supply
75W220/75W110
Epi High-intensity
Collector Lens
L
Filter Turrets
TI-DF Darkfield
Condenser Adapter
L
TI-FLEW-E Motorized
Excitation Filter Wheel*1
D
HA
I
C-FC Epi-fl
Collector Lens
Epi-fl Collector
Lens Q2
AN
B
Mercury Lamp Socket
Xenon Lamp
M
TI-DH
Stage Base
E
A
C-SHG1 HG
Starter 100-240
HG Lamphouse
HMX-3B/HMX-4B
TI-ND6-E Motorized
TI-ND6 Sextuple TI-N6 Sextuple
TI-ND6-PFS Perfect Sextuple DIC Nosepiece*1 DIC Nosepiece
Nosepiece
Focus with Motorized
Sextuple DIC Nosepiece
TI-C-TPH Phase Ring for
PH Unit 60x/PH3, 60x/PH4,
100x/PH3, 100x/PH4, 40x/PH3
Ti-E/B main body with bottom port only
C-Mount TV Adapter A
TI-TIRF
Stage-Up Lens*5
H
F
C-Mount Camera
TI-DS 30W
30W
6V
HALOGEN
G
K
TI-BP-EX Back Port Extension Kit*6
MA Halogen
Lamp 6V30W
JAPAN
A
DIC Sliders
CFI Objectives
Filter ø33 mm: GIF,
Heat Absorbing, NCB11,
ND2 A, ND16 A
V2-A LL Halogen
Lamp 12V100W
Filter ø45 mm:
GIF, Heat Absorbing,
NCB11, ND2 A, ND16 A
TI-C-LWD
LWD Lens
MC-TMD2 TI-C-CLWD
ELWD Lens CLWD Lens
Darkfield
Lens (Oil)
TI-FLC-E Motorized
Epi-fl Filter Turret*1
TI-FLC-E/HQ Motorized
Epi-fl Filter Turret*1
C-HGFIA
HG100W Adapter
TI-TIRF
Stage-Up Lens*5
Darkfield
Lens (Dry)
C-HGFIF15/30
C-HGFI HG
HG Fiber 1.5 m/3.0 m Fiber Illuminator
“Intensilight”
C-LHGFI HG Lamp
N
E
K
B
F
TI-TIRF TIRF
Illuminator Unit*4
Stages
Ti-U/B main body with bottom port only
F
65
S
54
50
45
40
35
30
C-HGFIE Motorized
HG Fiber Illuminator
“Intensilight”
Side Port
C-HGFIE-C
HG Controller
MADE IN CHINA
C-HSG Slide
Glass Holder
C-HT
Terasaki Holder
C-HU
Universal Holder
35 mm Petri
Dish Holder
TI-SH-U Universal Holder
TI-SH-W Well Plate Holder
TI-SH-J Stage Ring Holder
ENG-Mount
Camera
Reflection Mirror*7,
TIRF 60/40 Half Mirror
ENG-Mount
TV Adapter
A
B
L
D
Glass Stage Ring
32 mm
1x Relay
Lens N
TE Acrylic Stage Ring
I
TI-SAM Attachable
Mechanical Stage
C1 Single Mode Fiber
T-BPA
Photo
Adapter
Dedicated
Straight
Tube
TI-FLBW-E
Motorized
Barrier Filter
Wheel*1
TE-AT Dual
CCTV Adapter
Dedicated
C-Mount Adapter
H?N?L
E J
TI-TIRF
Stage-Up Lens*5
Full-mirror Block
C-LU2 2-Laser Board S
TI-TIRF-E Motorized TIRF
Illuminator Unit*1*4
Ti Laser Safety Kit
F
RS-232C
TI-SR
Rectangular Stage
M
TI-SR/F
Rectangular Stage with
Front Positioned Knob
TI-S-E Motorized
Stage*1
TI-SSR Short-handle
Rectangular Stage
TI-SP Plain Stage
LASER UNIT
LASER SHUTTER
INPUT
12V
1A
SAFTY
COVER
CLOSE
BINO
CLOSE
POWER
C-Mount
Camera
TI-S-ER Motorized
Stage with Encoders*1
TI-LUSU
C-Mount 0.7x C-Mount TV Adapter
VM2.5x
Relay Lens
Side Port
Tube
Reflection Mirror*7,
TIRF 60/40 Half Mirror,
PA-GFP DM Mirror
TI-LUSU Shutter Unit
TI-TIRF
Stage-Up Lens*5
N
TI-S-CON Motorized
Stage Controller*1
T-SHN Stage
Handle Knob
C-Mount TV Adapter
VM4x
L100 is available as option
D-SLR Camera
(1x format)
B
K
H N
Bottom Port
J
Communication Hub Units/Controllers
Analyzer
TI-PAU Photo Activation
Illuminator Unit*4
[email protected]
H?N?L
F
C-LU3 3-Laser Board
C-Mount
TV Adapter A
D-Series F-mount TI-BTY F-TY Tube
Adapter
I
Stage Riser
C-LU3EX 3-Laser Board
Reflection Mirror*7,
TIRF 60/40 Half Mirror,
PA-GFP DM Mirror
K
rp_lqnmpr_rgml,
dmsp`jmaiqbspgle
Dgvrfcj_qcpslgrugrf
H
TI-HUBC/A Hub
Controller A
C-Mount TV Adapter VM2.5x
C1 Single Mode Fiber
Ex Fine
Coarse
Coarse
Fine
TI-AC/A AC Adapter
for HUB-A
TI-AC100/120/230
Power Cord
XY Speed
Constant Speed
Fine
Ex Fine
+Y
T-A2 DIC
Analyzer
Z Speed
+X
-X
-Y
C-Mount TV Adapter VM4x
TI-AC/B AC Adapter
for HUB-B
J
TI-ERGC Ergo
Controller*1
TI-RCP Remote
Control Pad*1
TI-HUBC/B Hub
Controller B
C-Mount TV Adapter A
C-Mount Camera
TI-AC100/120/230
TI-S-EJOY
Power Cord
Stage Joystick*1
TI-BSUK70 70 mm
Stage Up Kit
I
TI-TIRF-PAU
TIRF-Photoactivation
Illuminator Unit*4
TI-TIRF
Stage-Up Lens*5
RG+JS2QS
J?QCP
CKGQQGML
NMUCP
TI-LU4SU Shutter Unit LU4
LU4A 4-Laser Module A
LU4-B5 Beam Splitter 50/50
N
*1: Requires a Communication Hub Controller *2: Cannot be used with stage riser *3: Combination with C-HGFI/HGFIE Fiber Illuminator “Intensilight” is not recommended *4: Cannot be attached to Ti-S
*5: Necessary for incorporating an illuminator unit in lower tier of the stratum structure *6: Necessary for Back Port Unit when used with TIRF-Photoactivation Illuminator Unit *7: Included in the Illuminator Units
22
23
Specifications
Main body
Port
Ti-E, Ti-E/B
Ti-U, Ti-U/B
Ti-S, Ti-S/L100
4
Ti-E: eyepiece 100%, left 100%, right 100%,
eyepiece 20%/left 80%
Ti-E/B: eyepiece 100%, left 100%, right 100%,
bottom 100%
Motorized port switching
4
Ti-U: eyepiece 100%, left 100%,
right 100%, optional
Ti-U/B: eyepiece 100%, left 100%,
right 100%, bottom 100%
Manual port switching
2
Ti-S: eyepiece 100%,
eyepiece 20%/left 80%
Ti-S/L100: eyepiece 100%, left* 100%
Manual port switching
*Changeable to right as option.
Two ports (tube base unit with side port, back port) can be added optionally.
Focusing
Intermediate
magnification
Eyepiece
tube
Via motorized nosepiece up/down movement
Stroke (motorized): up 7.5mm, down 2.5mm
Motorized (pulse motor)
Minimum step: 0.025µm
Maximum speed: 2.5mm/sec or higher
Motorized escape and refocus mechanism (coarse)
Coarse/fine switchable
Via nosepiece up/down movement
Stroke (manual): up 8mm, down 3mm
Coarse stroke: 5.0mm/rotation
Fine stroke: 0.1mm/rotation
Minimum fine reading: 1µm
Coarse refocusing mechanism
—
1.5x
—
Other
Light intensity control, Light on/off switch,
VPD on front of body, Operation with controller
Eyepiece tube body
TI-TD Binocular Tube D, TI-TS Binocular Tube S, TI-TERG Ergonomic Tube
Eyepiece tube base
TI-T-B Eyepiece Tube Base Unit, TI-T-BPH Eyepiece Tube Base Unit for PH, TI-T-BS Eyepiece Tube Base Unit with Side Port
Eyepiece lens
CFI 10x, 12.5x, 15x
—
Illumination pillar
TI-DS Diascopic Illumination Pillar 30W, TI-DH Diascopic Illumination Pillar 100W
Condenser
ELWD condenser, LWD condenser, NAMC condenser, ELWD-S condenser, High NA condenser, Darkfield condenser, CLWD condenser
Nosepiece
TI-ND6-E Motorized Sextuple DIC Nosepiece, TI-N6 Sextuple Nosepiece, TI-ND6 Sextuple DIC Nosepiece,
TI-ND6-PFS Perfect Focus with Motorized Sextuple DIC Nosepiece
Objectives
CFI60 objectives
Stage
TI-S-ER Motorized Stage with Encoders, TI-S-E Motorized Stage—Cross travel: X110 x Y75mm, Size: W400 x D300mm (except extrusions)
TI-SR Rectangula Stage, TI-SR/F Rectangular Stage with front positioned knob, TI-SSR Short-handle Rectangular Stage—
Cross travel: X70 x Y50mm, Size: W310 x D300mm
TI-SP Plain Stage—Size: W260 x D300mm
TI-SAM Attachable Mechanical Stage—Cross travel: X126 x Y84mm when used with TI-SP Plain Stage
Motorized functions
Focusing, Port switching, Coarse focusing
Epi-fluorescence attachment
Sextuple fluorescence filter cube rotating turret, Filter cubes with noise terminator mechanism,
Field diaphragm centerable, 33mm ND4/ND8 filters, 25mm heat absorbing filter
Option: Motorized sextuple fluorescence filter cube rotating turret, Motorized excitation filter wheel, Motorized barrier filter wheel
Nomarski DIC system
Contrast control: Senarmont method (by rotating polarizer)
Objective side prism: for individual objectives (installed in nosepiece)
Condenser side prism: LWD N1/N2/NR (Dry), HNA N2/NR (Dry/Oil) types
Weight (approx.)
Phase contrast set: 41.5kg
Epi-fl set: 45.4kg
Phase contrast set: 38.5kg
Epi-fl set: 42.3kg
Power consumption (max.)
Full set (with HUB-A and peripherals): approx. 95W
Full set (with HUB-B and peripherals): approx. 40W
—
Phase contrast set: 29.6kg
Epi-fl set: 33.4kg
24
Nikon’s Inverted Microscope Legacy
and the History of Discovery
2007
● Eclipse Ti-E, the next generation of
discoveries begins today
● PFS (perfect focus system)
● Laser TIRF
● Simplified DNA sequencing on the TE2000
2000
● Eclipse TE2000
● IR laser trapping
Eclipse Ti-E
● Special inverted model used in space
● Cumulina the mouse cloned on the TE300
1996
● Eclipse TE300
● Breakthroughs: CFI 60 optics expanded infinity space
● Dolly the sheep cloned on the Diaphot 300
● First intracytoplasmic sperm injection (ICSI) on the Diaphot
Eclipse TE2000
1990
● Diaphot 300
● High NA DIC
● Rectified DIC
Eclipse TE300
● Extra long working distance optics
● The brightest fluorescence
● World’s first IVF baby on the Diaphot TMD
1980
● Diaphot TMD, a revolutionary market leader
for inverted microscopy
● Beginning of FURA/CA+ 340nm imaging
Diaphot 300
1976
● First CF optics
● First Hoffman Modulation Contrast®
1966
● Model MSD, the first affordable tissue
culture microscope
1964
● Model M, the legacy begins
Diaphot TMD
● Pioneering 16mm time-lapse live cells
Model MSD
Model M
● Landmark achievements for Nikon
● Nikon’s unique technical innovations in inverted microscopy
● Key scientific breakthroughs and Nikon’s participation in some of these
Dimensional Diagram
169.5
169.5
169.5
196
196
260
260
260
172
153
449 (PD=64)
615
163.5
449 (PD=64)
452 (PD=64)
729
163.5
725
196
172
497
172
497
497
635
Unit: mm
Enter the “Microscopy University” on the web
and discover a whole new world.
www.microscopyu.com
Specifications and equipment are subject to change without any notice or obligation on
the part of the manufacturer. March 2010 © 2007-10 NIKON CORPORATION
WARNING
TO ENSURE CORRECT USAGE, READ THE CORRESPONDING
MANUALS CAREFULLY BEFORE USING YOUR EQUIPMENT.
Monitor images are simulated.
Company names and product names appearing in this brochure are their registered trademarks or trademarks.
NIKON CORPORATION
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phone: +81-3-3216-2375 fax: +81-3-3216-2385
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CHINA phone: +86-21-5836-0050 fax: +86-21-5836-0030
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Printed in Japan (1003-05)T
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SINGAPORE phone: +65-6559-3618 fax: +65-6559-3668
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UNITED KINGDOM phone: +44-208-247-1717 fax: +44-208-541-4584
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