Evanescent Wave Imaging System
imageVisualization 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.
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.
Image courtesy of:
Dr.Takahiro Fujiwara, Kusumi Membrane Organizer Project, ERATO, JST
An example of simultaneous mounting?configuration with epi-fluorescence attachment
TIRF objective is adjustable to correct for temperature changes
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.
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)
Images and explanations: Shuichi Obata, Ph.D., Department of Anatomy, School of Medicine, Yokohama City University
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)