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Scanning Confocal Microscope A1R+

Scanning Confocal Microscope  – Bring imaging to life: high-speed, high-resolution confocal imaging.

Capturing high-quality confocal images at ultrahigh-speed and enhanced sensitivity with a resonant scanner and galvano scanner, Nikon’s A1R+ is a powerful tool for the imaging and visualization of intracellular dynamics and interaction.

ER: Enhance Your Confocal Resolution

The A1 ER enhanced resolution Module combines powerful GPU-based processing and specialized deconvolution algorithms to dramatically enhance the spatial resolution of your A1+ or A1R+ confocal microscope with minimal processing time.

The A1 ER Module provides high quality PSF models for Nikon’s high performance objective lenses, taking the guesswork out of deconvolution analysis.  Other features include automatic or manual mode for iteration selection, enhanced spherical aberration correction, and robust algorithms for noise estimation and removal.

 

Revolutionary Hybrid Confocal Scan Head

The A1R+ has a hybrid scanner head that incorporates both an ultrahigh-speed resonant scanner and a high-resolution galvano scanner. Simultaneous photoactivation and ultrafast imaging using these two scanners allow acquisition of rapid changes after photoactivation and enables observation of intermolecular interaction.

A1R-Plus-Hybrid-Scanner
HeLa cells expressing Yellow Cameleon 3.60 were excited with 457 nm laser light. After stimulation with histamine, calcium ion concentration dynamics were observed. The (blue) emission of CFP and the (yellow) emission of YFP are shown as green and red channels respectively. The graph displays fluorescence intensity (vertical) versus time (horizontal). The green and red lines in the graph indicate the intensity change of CFP emission (green) and YFP emission (red) from the region of interest (ROI). Along with the increase of calcium ion concentration in the cell, the intermolecular FRET efficiency between CFP and YFP within Yellow Cameleon 3.60 increases, the CFP fluorescence intensity decreases, and the YFP fluorescence intensity increases. Imaging laser wavelength: 457 nm, Image size: 512 x 512 pixels, 30 fps (with resonant scanner) Photos courtesy of: Dr. Kenta Saito and Prof. Takeharu Nagai, Research Institute for Electronic Science, Hokkaido University

 

Resonant Scanner Provides Ultrafast Imaging

The A1R+ resonant scanner has an ultrahigh resonance frequency of 7.8 kHz. It allows imaging of intercellular dynamics at 30 fps (512 x 512 pixels) and 420 fps (512 x 32 pixels), the world’s fastest image acquisition. The field of view of the scanned area is approximately five times larger than that of the galvano scanner. The Nikon original optical clock generation method realizes high image quality even at the highest speed. The fiber-optic communication data transfer system can transfer data at a maximum of 4Gbps.

Mouse-Blood-Vessel
Mouse blood vessel administered Tetramethyl Rhodamine and Acridine Orange and observed at 120 fps (8 ms/frame, with resonant scanner) Red: blood vessel, Green: nucleus Tile images displayed every 8 ms The arrows indicate white blood cell flow in the vessel. Photos courtesy of: Dr. Satoshi Nishimura, Department of Cardiovascular Medicine, the University of Tokyo, Nano-Bioengineering Education Program, the University of Tokyo, PRESTO, Japan Science and Technology Agency

Galvano Scanner Enables High-resolution Imaging

The A1+ utilizes a galvano scanner which enables high-resolution imaging of up to 4096 x 4096 pixels. In addition, with the newly developed scanner driving and sampling systems, plus image correction technology, high-speed acquisition of 10 fps (512 x 512 pixels) is also possible.

Zebrafish labeled with four probes

Zebrafish labeled with four probes (captured with galvano scanner) Nucleus (blue): Hoechst33342, Pupil (green): GFP, Nerve (yellow): Alexa555, Muscle (red): Alexa647.

Drosophila sp. embryonic heart

Drosophila sp. embryonic heart

bovine_brain

Bovine brain microvascular endothelial cells labeled with MitoTracker (mitochondria, yellow), phalloidin (actin, blue) and Hoechst (DNA, magenta).

GaAsP Multi Detector Unit

Nikon developed the GaAsP multi-detector unit equipped with two GaAsP PMTs and two normal PMTs.

A GaAsP PMT has much higher sensitivity than a normal PMT, thus acquisition of brighter signals with minimal background noise is possible with a GaAsP PMT, even with weak fluorescence, which, until now, has been difficult to detect.

When using resonant scanners, the GaAsP PMT enables low-noise, high-speed imaging.

GaAsP-PMT
Sensitivity comparison of GaAsP PMT and normal PMT GaAsP PMT realizes higher sensitivity than normal PMT, thus offering high quantum efficiency up to 45%. * Quantum efficiency indicates logarithm
gaasp-detector

GaAsP detector

normal-detector

Normal detector

Increased Light Detection Efficiency

The low-angle incidence method utilized on the dichroic mirrors increases fluorescence efficiency by 30%.

Conventional 45°
incidence angle method
conventional-45

Reflection-transmission characteristics have high polarization dependence Low-angle
incidence method

low-angle

Reflection-transmission characteristics have lower polarization dependence

By employing the hexagonal pinhole, higher brightness equivalent to that of a circular pinhole is achieved.

square-pinhole

Square pinhole

64% of the area of the circle

30% more light
pinhole-arrow
hexagonal-pinhole

Hexagonal pinhole

83% of the area of the circle

Nikon’s original dual integration signal processing technology (DISP) has been implemented in the image processing circuitry to improve electrical efficiency, resulting in an extremely high S/N ratio.

Enhanced Spectral Imaging

Acquisition of a 32-channel spectral image (512 x 512 pixels) with a single scan in 0.6 second is possible. Moreover, 512 x 32-pixel images can be captured at 24 fps.

Accurate, High-speed Unmixing

Accurate spectral unmixing provides maximum performance in the separation of closely overlapping fluorescence spectra and the elimination of autofluorescence. Superior algorithms and high-speed data processing enable real time unmixing during image acquisition.

spectral_unmixing
Spectral and unmixed images of five-color-fluorescence-labeled HeLa cells. Specimen courtesy of: Dr Tadashi Karashima, Department of Dermatology, Kurume University School of Medicine

V-filtering Function

Filter-less intensity adjustment is possible by selecting desired spectral ranges from 32 channels that match the spectrum of the fluorescence probe in use and combining them to perform the filtering function.

V-Filtering

Increased flexibility and ease of use

NIS-Elements C control software enables integrated control of the confocal imaging system, microscope and peripheral devices with a simple and intuitive interface. Diverse reliable analysis functions are also available.

Combine with Super-Resolution Modalities

Multi-mode imaging is possible by equipping the A1R+ with Nikon’s N-SIM E or N-STORM 4.0 super-resolution system. Combining confocal and super-resolution modalities on the same microscope enables users to easily compare and verify super-resolution data with traditional confocal images.  In addition, users can take full advantage of the confocal imaging modality to acquire contextual information for their super-resolution images.

N-SIM E

Nikon N-SIM E Super-Resolution System

SPECIFICATIONS

Scan head input/output port 2 laser input ports 3 signal output ports for standard, spectral and optional detector*1
Laser LU-N3 3-laser unit 405 nm, 488 nm, 561nm lasers are installed ; built-in AOTF *Cannot be used with spectral detector
LU-N4/LU-N4S
4-laser unit
405 nm, 488 nm, 561nm,640nm laser are installed;built-in AOTF *Use LU-N4S when using the spectral detector
LU-NV series laser unit Compatible lasers : 405 nm, 445 nm, 458nm,488nm,514nm,532nm,561nm,594nm,640nm,647nm ; built-in AOTF
Standard fluorescence detector Wavelength 400-750 nm
Detector A1-DU4 4 Detector Unit: 4 standard PMTs A1-DUG GaAsP Multi  Detector Unit: 2 GaAsP PMTs + 2 standard PMTs
Filter cube 6 filter cubes commonly used for a microscope mountable on each of three filter wheels Recommended wavelengths: 450/50, 482/35, 515/30, 525/50, 540/30, 550/49, 585/65, 595/50, 700/75
Diascopic detector (option) Wavelength 485-650 nm
Detector PMT
FOV Square inscribed in a ø18 mm circle
Image bit depth 4096 gray intensity levels (12 bit)
Scan head Standard image acquisition Scanner: galvano scanner x2 Pixel size: max. 4096 x 4096 pixels Scanning speed: Standard mode: 2 fps (512 x 512 pixels, bi-direction), 24 fps (512 x 32 pixels, bi-direction) Fast mode: 10 fps (512 x 512 pixels, bi-direction), 130 fps (512 x 32 pixels, bi-direction)*2 Zoom: 1-1000x continuously variable Scanning mode: X-Y, X-T, X-Z, XY rotation, Free line
High-speed image acquisition Scanner: resonant scanner (X-axis, resonance frequency 7.8 KHz), galvano scanner (Y-axis) Pixel size: max. 512 x 512 pixels Scanning speed: 30 fps (512 x 512 pixels) to 420 fps (512 x 32 pixels), 15,600 lines/sec (line speed) Zoom: 7 steps (1x, 1.5x, 2x, 3x, 4x, 6x, 8x) Scanning mode: X-Y, X-T, X-Z Acquisition method: Standard image acquisition, High-speed image acquisition, Simultaneous photoactivation and image acquisition
Dichroic mirror Low-angle incidence method, Position: 8 Standard filter: 405/488, 405/488/561, 405/488/561/638, 405/488/543/638, 457/514, BS20/80 Optional filter: 457/514/561
Pinhole 12-256 µm variable (1st image plane)
Spectral detector*3
(option)
Number of channels 32 channels
Wavelength detection range 400-750 nm
Spectral image acquisition speed 4 fps (256 x 256 pixels), 1000 lps Pixel size: max. 2048 x 2048
Wavelength resolution 80 nm (2.5 nm), 192 nm (6 nm), 320 nm (10 nm) Wavelength range variable in 0.25 nm steps
Unmixing High-speed unmixing, Precision unmixing
Z step Ti-E: 0.025 µm, FN1 stepping  motor:  0.05 µm Ni-E: 0.025 µm
Compatible microscopes ECLIPSE Ti-E inverted microscope, ECLIPSE FN1 fixed  stage  microscope, ECLIPSE Ni-E upright microscope (focusing nosepiece type and focusing stage type)
Option Motorized  XY stage (for Ti-E/Ni-E), High-speed Z stage (for Ti-E), High-speed piezo objective-positioning  system (for FN1/Ni-E)
Software Display/image generation 2D analysis, 3D volume rendering/orthogonal,  4D analysis, spectral unmixing
Image format JP2, JPG, TIFF, BMP, GIF, PNG, ND2,  JFF, JTF, AVI, ICS/IDS
Application FRAP, FLIP, FRET(option), photoactivation, three-dimensional time-lapse imaging, multipoint time-lapse imaging, colocalization
Control computer OS Microsoft Windows® 7 Professional 64bits SP1
CPU Intel Xeon E5-2643v3 (3.40 GHz/20 MB) or higher
Memory 16 GB or higher
Hard disk 300 GB SAS (15,000 rpm) x2, RAID 0 configuration
Data transfer Dedicated data transfer I/F
Network interface 10/100/1000 Gigabit Ethernet x2
Monitor 1600 x 1200 or higher resolution, dual monitor configuration recommended
Recommended installation  conditions Temperature 23 ± 5 ºC, humidity  70 % (RH) or less (non-condensing)

* 1 FCS/FCCS/FLIM is possible in combination with third-party systems

* 2 Fast mode is compatible  with  zoom 8-1000x and scanning modes X-Y and X-T. It is not compatible with Rotation, Free line,  CROP, ROI, Spectral  imaging, Stimulation and FLIM.

* 3 Compatible with galvano scanner only.