At present, OLYMPUS has launched the third generation of Total Internal Reflection Fluorescence Microscopy (TIRFM) illumination system. Since OLYMPUS launched the world's first commercial TIRFM illuminator in 1998, the latest development has further strengthened our leading position in this field. High numerical aperture apochromatic objectives 100x / 1.65NA and 60x / 1.45NA complete our system. TIRFM is a very special optical technology that has been widely used by cell biologists and neuroscientists. Using this technology, you can observe single molecules and study cell membranes.
TIRFM technology relies on the extremely shallow evanescent waves generated by oblique rays on the surface of two different refractive index optical media. The effect of this effect is that the refractive index of the incident medium is greater than the refractive medium, and the incident angle is greater than the critical angle of total reflection when the oblique radiation hits the optical interface. The residual evanescent wave diffuses into the low-refractive-index medium, and the light intensity decreases exponentially as the diffusion depth increases. The extremely shallow transmissivity of the evanescent wave can be used to illuminate and excite fluorescent dyes within 100nm of the optical surface. Therefore, the autofluorescence interference in the cell is almost completely eliminated. Compared with traditional fluorescent lighting technology, the image information Noise ratio.
Olympus was the first company to develop a TIRFM system into a commercial product. The latest generation of TIRFM illuminators can provide 3 different wavelengths of multispectral line illumination simultaneously through optical fibers. No need to modify the body of the IX2 microscope when installing the illuminator. The built-in micrometer can easily adjust the laser illumination.
Our objective TIRFM system supports traditional reflective and transmission wide-field observations such as brightfield, darkfield, phase contrast, and differential interference contrast (DIC) observations. The objective TIRFM system is different from the prism TIRFM system. The prism TIRFM system requires an expensive prism system to be installed above the specimen, and the installation of the prism will restrict the transmitted light observation technology and cause specimen vibration. The core of the objective TIRFM system technology is the unique apochromatic TIRFM objective lenses that OLYMPUS prides itself on, including two types of 100x and 60x objectives with numerical apertures of 1.65 and 1.45, respectively. Using a 100X TIRFM objective lens, the brightness of its brightfield fluorescence microscope is 28% higher than that of a conventional 100Xoil / 1.4NA objective lens, and the resolution is also improved from 0.240 μm to 0.203 μm. This high numerical aperture TIRFM objective lens optimizes both the adjustment of the effective operating surface and the adjustment of the laser illumination angle. In addition, TIRMF objectives are also suitable for other reflected light technologies.
| Sanford M. Simon et. Al. |
The Rockefeller University Laboratory of Cellular Biophysics
| Green: Vamp7-CFP |
Three high numerical apertures designed for evanescent wave lighting
100X NA: 1.65
极 Extremely high numerical aperture provides very bright fluorescent images sheet
100X NA: 1.45
更高 Higher cost performance and convenient operability Using standard mirror oil and coverslips can be used
60X NA: 1.45
更 More affordable price and more convenient operation 可以 Compatible with high-contrast and high-resolution DIC prisms Variations Use standard mirror oils and coverslips
The laser is directed into the microscope through a single mode fiber. Use the linkage mode to place the laser light out of the microscope or stimulate the eye through the eyepiece.
Introduce multiple lasers <br style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; padding-top: 0px"> 3 lasers can be introduced at the same time through the optional laser integrator .
Laser can be used:
Argon 488nm, HeNe-R 633nm, HeNe-G 543nm
Laser light adjustment with simple adjustments <br style="padding-bottom: 0px; margin: 0px; padding-left: 0px; padding-right: 0px; padding-top: 0px"> For single mode fiber After the initial adjustment of the laser head and the TIRFM illuminator, the subsequent adjustment only needs to simply adjust the micrometer to change the TIRFM lighting.
You can use an arc light to observe the fluorescence
The TIRFM illuminator can be connected to a standard arc lamp illuminator simultaneously, allowing simultaneous observation of evanescent waves and standard illumination fluorescence.