Microscopes

The Nikon A1R Confocal System

Nikon A1R

The A1R is the high-end confocal solution from Nikon. Our system is equipped with 4 laser lines and 4 separate photomultiplier units for 405nm, 488nm, 561nm and 640nm wavelengths and a spectral detector as well. The detector unit contains several dichroic mirrors and emission filters that can be changed using motorized filter wheels. The scanning head contains a hybrid scanner system incorporating both galvano and resonant scanners giving the user the extra option of high speed data acquisition. Using the resonant scanner one can scan a 515x512 pixel area at 30 frames per second. The two scanners can even be used simultaneously in photoactivation/photobleaching experiments. In these cases the galvano scanner is used for the activation of a given area and the resonant scanner is used for imaging. A separate PMT can be used to detect transmitted light. This can also be used to create DIC images with a polarizing DIC filter cube, a Nomarski prisms in the condenser/under the objective, and an addition polarizer. The scanner is equipped on a fully motorized Nikon Ti Eclipse inverted microscope with high quality plan fluor and plan apo objectives (4X, 10x,20x,40x,60x(Water), 60x(Oil)). Fluorescent episcopic illumination is achieved using the Intensilight illumination system and a set of DAPI, FITC and TRIC filters. The setup is well suited for live cell imaging as it is outfitted with a SOLENT environmental chamber with controlled temperature, humidity and CO2 atmosphere. Overall this system is a very advanced tool and using it's the motorized components in combination with the JOBS module of the NIS Elements software package.

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The Nikon C2 Confocal System

Nikon C2

The C2 plus microscope is Nikon’s “work horse” laser scanning microscope. The scanning unit is equipped on a fully motorized Nikon Eclipse (Ni-E) microscope body. Our C2 confocal system enables users to acquire up to four channel images (available laser lines are: 405nm, 488nm, 561nm and 640nm) using 3 highly sensitive photomultiplier (PMT) detectors. Besides the confocal mode users can use the microscope in widefield mode with the Nikon Intensilight illuminator, a Nikon DS-Fi3 5.9 MP color CMOS camera and DAPI, FITC and TRIC filter cubes. The extremely precise motorized stage along with the motorized objective turret and filter wheel allow users to devise advanced imaging schemes. The microscope is equipped with high quality plan fluor and plan apo objectives (4X, 10x,20x, 60x). This very robust microscope is very user friendly and is suited for most biological applications.

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The Nikon N-STORM System

Nikon N-STORM

The Nikon N-STORM system is one of the crown jewels among Nikon’s microscope systems. Stochastic Optical Reconstruction Microscopy (STORM) is a revolutionary technique that achieves super-resolution by temporally separating the registration of individual fluorophores. The most used case is the application of photo-switchable dyes that have a reversible dark state in the appropriate medium. A powerful laser is used to drive almost all of these molecules into a dark state. Periodically a few molecules return to the ground state and due to continuous illumination is excited again and again until it reaches the dark state again. Recording these spatially separated blinking events and fitting the centroid of the Point Spread Function (PSF) results in a point list. This point list will ulteriorly outline the investigated structure and enables the system to reach about tenfold increase in lateral/axial resolution (20-60nm laterally and 50-100nm axially).

The microscope is a TIRF microscope equipped with a series of lenses that focus the incoming laser onto the specimen to further increase the local light intensity 4x or 8X. Additionally it is equipped with Nikon’s cutting edge Perfect Focus System that compensates for drift in the Z direction. The unsurpassable stability achieved thanks to the PSF system is indispensable for STORM imaging. The microscope laser board contains 4 lines: 405,488,561 and 642nm (out of which the 642 and 561 are high power lasers) and a series of high quality filter cubes (HQ Blue, HQ Green, HQ Red, Far-red STORM and Red/Far-Red Dual STORM cube). The microscope is also fitted with a C2 confocal head that allows correlative STORM-Confocal imaging. Users can choos between 10x and 20x Plan Fluor and a 100x TIRF Plan Apo objective.

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The Nikon Fn1 A1R Multiphoton System

Nikon A1RMP

Multiphoton microscopes are used for in-vivo imaging where samples are thick. Multiphoton microscopy uses two-photon excitation for imaging the sample. For a given transition in two-photon excitation fluorophores absorb two photons with half the energy required for single photon excitation. The probability of two-photon excitation is low and is proportional to the square of the excitation light. Two photon microscopy uses exactly this fact to achieve optical sectioning. If a high power laser is focused in the sample two photon excitation will only happen in a small volume around the focal point. As opposed to confocal microscopy where fluorophores are excited in a much larger volume and a pinhole is needed to filter off focus fluorescence, in multiphoton microscopy the pinhole can be omitted (non-descended detectors) as off-focus dyes are not excited. As the energy of the photons involved is half the laser wavelength should be double. This means that the laser wavelength falls in the IR range where the tissue absorbs and scatters less.

Our multiphoton setup is optimized for physiological experiments in thick slices and is equipped for single cell electrophysiology (whole-cell, outside-out, inside-out and perforated patch). The microscope is equipped with the Nikon A1RMP+ scanning head that allows high speed scanning from 30 fps at 512x512 pixels to 420 fps in line scanning mode. This makes the setup a great tool to follow Ca2+ transients in real time. The microscope uses a Coherent Chameleon Vision II Ti:Sapphire laser that can be tuned between 700 nm and 1200 nm-s. Nikon’s laser incident unit has an auto-alignment mechanism that automatically corrects. The detection unit of the microscope has 2 highly sensitive GaAsP and 2 standard NDD detectors. The microscope uses an outstanding Nikon 25x Apochromat long working distance objective that has an exceptionally high numerical aperture of 1.1. For high speed Z-stacks the setup is equipped with a Mad City Labs Piezo scanner nosepiece. The setup is mounted on a Luigs and Neumann shifting stage with a three axis motorized micromanipulator. The micromanipulators are equipped with a headstages/pipette holder that is connected to a Multiclamp 700B amplifier and Digidata 1322A digitizer from Molecular Devices. Additionally the setup is equipped with a Hamamatsu CCD camera for diascopic imaging in the IR rang for patch clamp experiments.

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The Nikon Fn1 C1 Confocal System

Nikon C1

FN1-C1 configuration allows high-resolution imaging of deeper areas of cells with the excellent patch clamp operability and performance of NIKON objectives. The FN1 Eclipse was developed especially for electro-physiological research with features to facilitate patch-clamp experiments. Advantages offered by the FN1 include slim body, streamlined structure, improved electrode placement, long working distance, and greater noise reduction. The microscope is equipped a Nikon C1 scanning head and three available laser lines at 408nm, 488nm and 543nm. Additionally the setup is equipped with an Andor Zyla sCMOS for diascopic imaging in the IR rang for patch clamp experiments. The microscope uses an outstanding Nikon 25x Apochromat long working distance objective that has an exceptionally high numerical aperture of 1.1. This is a complete physiological setup that can be used for field and pair recordings as well. The micromanipulator system is made up of a Luigs and Neumann shifting stage, a stage with a bath chamber, two micromanipulators with three motorized axes, a 3 axis manual manipulator and a Kleindiek micromanipulator with nanometer resolution. Two micromanipulators are equipped with two headstages/pipette holders that are connected to a Multiclamp 700B amplifier and Digidata 1440A digitizer from Molecular Devices.

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The Nikon Fn1 Andor Revolution Spinning Disk System

Nikon Andor Revolution

The Nikon FN1 microscope was developed especially for electrophysiological experiments. It is a simple and extremely robust microscope base streamlined to improved electrode placement, allow a long working distance and reduce noise. The system is a fully equipped electrophysiology station that uses high quality Nikon objectives, an Andor Revolution XD system with a Yokogawa CSU-X1 confocal scanner, Luigs and Neumann stage system (3 motorized axis for electrodes). The Andor Revolution® XD spinning disk laser microscopy system is a high speed spinning disk system that uses a microlens disk and Nipkow disk that act as pinholes (20 000 each). As the disk is rotating at any given time multiple lens/pinholes are irradiated with the lasers and the resulting image is projected onto a very sensitive Andor iXonEM+ EMCCD. This way multiple points can be scanned at once and the system can achieve 2000 fps. Additionally the system uses a high speed filterwheel that can change in 33ms. Currently the confocal laer board is equipped with two laserlines at 488nm and 561nm. The setup is also enabled for diascopic imaging in the IR range with DIC. The setup is equipped with a long working distanse 16x Water Dipping Achromat with extremely large Numerical Aperture and optimized for the IR range. This is a complete physiological setup that can be used for field and pair recordings as well. The micromanipulator system is made up of a Luigs and Neumann shifting stage, a stage with a bath chamber, three micromanipulators with three motorized axes. The three micromanipulators are equipped with headstages/pipette holders that are connected to a Multiclamp 700B amplifier and Digidata 1440A digitizer from Molecular Devices.

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