Visualization of single molecule fluorescence
in live cells
The Evanescent Wave Imaging System utilizes evanescent waves
to excite single molecules in the thin section in contact
with the coverglass.
Nikon's high N.A. TIRF objectives make it possible to introduce
laser illumination at incident angles greater than the critical
angle (θc) resulting in TIR that creates an evanescent
wave immediately adjacent to the coverglass-specimen interface.
The evanescent wave typically reaches less than 100nm into
the specimen and its energy drops off exponentially. Because
the specimen is not excited beyond the evanescent wave, this
imaging system can produce fluorescence images with an extremely
high signal-to-noise (S/N) ratio. |
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TIRF and other attachments
simultaneously mountable
The unique extendible "stratum structure” of Nikon's TE2000
inverted microscope permits simultaneous mounting of both
TIRF and epi-fluorescence illumination systems, with no restriction
on their individual capabilities. It also allows their dedicated
filters to be used independently. Moreover, switching between
the two systems is elementary. |
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Image
courtesy of:
Dr.Takahiro Fujiwara, Kusumi Membrane Organizer Project,
ERATO, JST |
An example
of simultaneous mounting?configuration with epi-fluorescence
attachment |
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TIRF objective is adjustable to correct
for temperature changes
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CFI Plan Apo TIRF 60X/1.45
(left)
CFI Plan Apo TIRF 100X/1.45 (right) |
Nikon has developed a dedicated 60X TIRF objective,
the first lens in the world that corrects for temperature
changes. By using a coverglass thickness correction collar,
you can easily correct temperature-induced changes-from 23'c
(room temperature) to 37'c (physiological temperature)-in
the refractive index of the immersion oil that can cause spherical
aberration. |
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SRIC (Surface Reflective Interference
Contrast) method can reveal focal contacts prior to switching
to TIRF
Under the SRIC method, only the areas in contact with the
coverglass are rendered in black within the specimen. With
Nikon's TIRF system, the SRIC method can be easily used by
changing the combination of epi-fl filters. This makes it
possible to check whether or not the specimen should be visible
by TIRF before fluorescence excitation. This eliminates the
risk of photo-bleaching during focusing. Then, you can easily
switch to TIRF observation since both TIRF and epi-illumination
systems are simultaneously mounted.
(Patent pending) |
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Images and explanations: Shuichi
Obata, Ph.D., Department of Anatomy, School of Medicine, Yokohama
City University |
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Main Specifications |
*Microscope
Unit: |
Nikon
Inverted Microscope TE2000 (with epi-fluorescence
attachment) |
Dedicated
objectives: |
CFI
Plan Apo TIRF 60X/1.45 oil,
CFI Plan Apo TIRF 100X/1.45 oil |
|
Direct
C-mount TV adapter |
*Evanescent
Wave Illumination Unit: |
Laser:
blue (488), green (532), red (633), up to 3 types
can be used |
Provided
with filter cassette (dichroic mirror/barrier filter) |
Aperture
diaphragm with adjustment mechanism |
ND filters
(ND2, ND8, ND32) |
|
data
sheet (*.pdf) |
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