Amazingly deep — A1 MP+/A1R MP+ sharply visualize ultra-deep dynamics within living organisms.
Configured with Ni-E
The A1 MP+/A1R MP+ multiphoton confocal microscopes provide faster and sharper imaging from deeper within living organisms, extending the boundaries of traditional research techniques in biological sciences. They are compatible with both upright and inverted microscopes, and provide optimum multiphoton imaging configurations for brain research, other neuroscience applications and in vivo imaging of living specimens.
The Nikon resonant scanner is equipped with A1R MP+ and capable of high-speed 420 fps (512 x 32 pixels) imaging, the world's fastest for a multiphoton microscope using point scanning technology. Unique to this design is a resonant scan mirror capability that enables imaging full fields of view at much higher speeds than with conventional galvano scanners. Nikon's optical pixel clock system, which monitors the position of the resonant mirror in real time, adjusts the pixel clock to ensure more stable, geometrically correct and more evenly illuminated imaging, even at high speeds. This enables the successful visualization of in vivo rapid changes, such as reactions in living organisms, dynamics and cell interactions.
The fluorescence emissions from deep within a specimen are highly scattered in multiphoton excitation, and therefore the conventional detector using a pinhole cannot provide bright fluorescent images. The episcopic NDD in the A1 MP+/A1R MP+ is located close to the back aperture of the objective to detect the maximum amount of scattered emission signals from deep within living specimens. The use of this four-channel detector in combination with special spectral mirrors, together with Nikon's unmixing algorithm, eliminates cross talk between fluorescent probes with highly overlapping emission spectra. Background auto-fluorescence is also eliminated, enabling high-contrast image capture from deep within the specimen. Using diascopic NDD* together with episcopic NDD, brighter images can be acquired by detecting fluorescence signals from both reflected and transmitted.
4-channel episcopic NDD
4-channel diascopic NDD
The newly developed gallium arsenide phosphide (GaAsP) NDD* has approximately twice the sensitivity of a standard NDD and allows clear imaging of deeper areas of in vivo mouse brain over 1.2mm. Its ability to acquire bright images enables faster imaging and higher quality Z-stack imaging. Its high sensitivity allows acquisition of fluorescent signals with less laser power, resulting in less photo damage to living specimens.
High-NA objectives have been developed that highly correct chromatic aberrations over a wide wavelength range, from ultraviolet to infrared. Transmission is increased through the use of Nikon's exclusive Nano Crystal Coat technology. In particular, the CFI Apochromat 25xW MP objective lens provides an industry leading highest numerical aperture of 1.10 while still maintaining a 2.0 mm working distance. It also has a collar that corrects chromatic aberrations depending on the depth of the specimen and a 33° manipulator pipette access angle, making it ideal for deep multiphoton imaging and physiology research applications.
CFI75 Apochromat 25xW MP
CFI Apochromat LWD 40xWI λS
CFI Apochromat 40xWI λS
CFI Plan Apochromat IR 60xWI
| CFI75 Apochromat 25xW MP | NA 1.1 WD 2.0 Nano Crystal Coat |
|---|---|
| CFI Apochromat LWD 40xWI λS | NA 1.15 WD 0.6 Nano Crystal Coat |
| CFI Apochromat 40xWI λS | NA 1.25 WD 0.18 Nano Crystal Coat |
| CFI Plan Apochromat IR 60xWI | NA 1.27 WD 0.17 Nano Crystal Coat |
When the multiphoton laser wavelength or group velocity dispersion pre-compensation is changed, the multiphoton laser beam positional pointing at the objective back aperture may also change, resulting in uneven intensity across the image, or a slight misalignment between the IR and visible laser light paths.
Verifying the IR laser beam pointing and setting the alignment has traditionally been difficult. Nikon's A1 MP+ series' auto laser alignment function, housed in the Incident Optical Unit for the multiphoton excitation light path, automatically maximizes IR laser alignments with a single click in NIS-Elements C.
A1R MP+ hybrid scanner
A1 MP+ equipped with a galvano (non-resonant) scanner enables high-resolution imaging of up to 4096 x 4096 pixels and high-speed acquisition of 10 fps (512 x 512 pixels). A1R MP+ is a hybrid scanning head that incorporates both a galvano scanner and an ultrahigh-speed resonant scanner. A1R MP+ allows imaging and photoactivation at ultrafast speeds at 420 fps (512 x 32 pixels) necessary for revealing cell dynamics and interaction.
Three color simultaneous fluorescent imaging with 850 nm pulsed IR excitation
(left: before unmixing, right: after unmixing)
