Super Resolution Microscope N-SIM E. Super Resolution Microscope

Similar documents
Microscope objectives

Confocal Microscope. Confocal Microscope C2

Super High Vertical Resolution Non-Contact 3D Surface Profiler BW-S500/BW-D500 Series

Super Resolution Microscope N-SIM/N-STORM. Super Resolution Microscope

Super Resolution Microscope. Super Resolution Microscope N-SIM/N-STORM

Confocal Microscope. Confocal Microscope C2

Biological Microscope. Biological Microscope ECLIPSE E100

Nikon. King s College London. Imaging Centre. N-SIM guide NIKON IMAGING KING S COLLEGE LONDON

Nikon Instruments Europe

Confocal NEXIV VMZ-K Series. CNC Video Measuring System CONFOCAL NEXIV. VMZ-K Series

Confocal NEXIV VMZ-K Series. CNC Video Measuring System CONFOCAL NEXIV. VMZ-K Series

CNC Video Measuring System NEXIV VMZ-K series. CNC Video Measuring System. Confocal Model

Wafer Loaders for IC Inspection Microscopes NWL200 Series. Wafer Loaders for IC Inspection Microscopes

Upright Microscope. Upright Microscope ECLIPSE Ci-E/Ci-L

Upright Microscope. Upright Microscope ECLIPSE Ci-E/Ci-L

Complete support to all your needs

Inverted Research Microscope. Inverted Research Microscope ECLIPSE Ts2R

Supplementary Information. Stochastic Optical Reconstruction Microscopy Imaging of Microtubule Arrays in Intact Arabidopsis thaliana Seedling Roots

Working Simultaneously. The Next Level of TIRF Microscopy. cell^tirf Illuminator Motorized Total Internal Reflection Fluorescence

The Next Level of TIRF Microscopy. cell^tirf Illuminator Motorized Total Internal Reflection Fluorescence

contents TABLE OF The SECOM platform Applications - sections Applications - whole cells Features Integrated workflow Automated overlay

Objectives for biological microscopes. Objectives for biological microscopes

High-resolution, low light-dose lightsheet microscope LATTICE LIGHTSHEET

L100 The ultimate CMM laser scanner combining productivity and accuracy NIKON METROLOGY I VISION BEYOND PRECISION

IC 2 S High Performance Objectives

OPELCO OPtical ELements COrporation LB Objective Series for Biological Use

Confocal Microscope A1+/A1R+ Confocal Microscope

Confocal Microscope A1+/A1R+ Confocal Microscope

Life Science Instrumentation. New Generation. Light Sheet Fluorescence Microscope. Alph

Very short introduction to light microscopy and digital imaging

Cell culture, observation and image capture three functions incorporated into one single long-term time-lapse imaging system

ImageXpress Micro XLS Widefield High Content Screening System. Imaging with a vision.

Leica_Dye_Finder :53 Uhr Seite 6 Dye Finder LAS AF

Lecture 16. OMX - Structured Illumination Microscopy Ian Dobbie x Microscopy Course Lecture 16 1

VMZ-K3040 CONFOCAL. Confocal Imaging & Metrology. CNC Video Measuring System. Specifications. Dimensional Diagram

Technology Note ZEISS LSM 880 with Airyscan

Multiphoton confocal microscope. Multiphoton confocal microscope A1R MP

Opterra II Multipoint Scanning Confocal Microscope. Innovation with Integrity

Introduction to Light Microscopy. (Image: T. Wittman, Scripps)

Why and How? Daniel Gitler Dept. of Physiology Ben-Gurion University of the Negev. Microscopy course, Michmoret Dec 2005

Image Analysis Software. Advanced Solutions for your Imaging World

Bio 407. Applied microscopy. Introduction into light microscopy. José María Mateos. Center for Microscopy and Image Analysis

INTRODUCTION TO OPTICAL MICROSCOPY

Microscopy Live Animal Imaging

Operation Guide for the Leica SP2 Confocal Microscope Bio-Imaging Facility Hunter College October 2009

Last updated: May 2014 Y.DeGraaf

attocfm I for Surface Quality Inspection NANOSCOPY APPLICATION NOTE M01 RELATED PRODUCTS G

LC15Dx High accuracy with high resolution

DeltaMyc. Fluorescence Lifetime Mapping Microscope. Affordable Fluorescence Lifetime Imaging Microscopy (FLIM)

Camera Overview. Digital Microscope Cameras for Material Science: Clear Images, Precise Analysis. Digital Cameras for Microscopy

IN Cell Analyzer 6500HS

microscopy A great online resource Molecular Expressions, a Microscope Primer Partha Roy

Diskovery Spinning Disk Guide

Confocal Laser Scanning Microscopy

FPD/LSI Inspection Microscopes

Introduction to light microscopy

Practical Flatness Tech Note

1 Co Localization and Working flow with the lsm700

CCAM Microscope Objectives

Microscope Objectives for Bioscience. Microscope Objectives for Bioscience

Imaging Introduction. September 24, 2010

Multifluorescence The Crosstalk Problem and Its Solution

Nature Protocols: doi: /nprot Supplementary Figure 1. Schematic diagram of Kőhler illumination.

Camera Overview. Digital Microscope Cameras for Material Science: Clear Images, Precise Analysis. Digital Cameras for Microscopy

Camera Overview. Digital Microscope Cameras for Material Science: Clear Images, Precise Analysis. Digital Cameras for Microscopy

We attempted to separate the two dyes by acquiring images using a single excitation wavelength and just two emission wavelengths.

Microscope Objectives for Bioscience. Microscope Objectives for Bioscience

Opterra. Multipoint Scanning Confocal Microscope. Innovation with Integrity. Cell-Friendly, High-Speed, High-Resolution Imaging

Rapid three-dimensional isotropic imaging of living cells using Bessel beam plane illumination

Point Spread Function. Confocal Laser Scanning Microscopy. Confocal Aperture. Optical aberrations. Alternative Scanning Microscopy

Nikon Eclipse Ti2-E Widefield/Spinning Disk Confocal Microscope Standard Operation Protocol

SHORT INSTRUCTIONS FOR OPERATING LSM1/2 (Zeiss LSM510) AT CIAN Version 1.4, September 2014

Specifications Optical system Main body Focusing. Dimensional Diagram. Fixed Stage Microscope for Electrophysiological Research

Application Note. The New 2D Superresolution Mode for ZEISS Airyscan 120 nm Lateral Resolution without Acquiring a Z-stack

Development of a High-speed Super-resolution Confocal Scanner

NeoScope. Simple Operation to 40,000. Table Top SEM. Serving Advanced Technology

Advanced Optical Microscopy lecture. 03. December 2012 Kai Wicker

ANSWER KEY Lab 2 (IGB): Bright Field and Fluorescence Optical Microscopy and Sectioning

picoemerald Tunable Two-Color ps Light Source Microscopy & Spectroscopy CARS SRS

Basics of confocal imaging (part I)

Nature Methods: doi: /nmeth Supplementary Figure 1. Schematic of 2P-ISIM AO optical setup.

3. are adherent cells (ie. cells in suspension are too far away from the coverslip)

Bringing Answers to the Surface

CNC Video Measuring System. CNC Video Measuring System NEXIV VMZ-R

Supplemental Figure 1: Histogram of 63x Objective Lens z axis Calculated Resolutions. Results from the MetroloJ z axis fits for 5 beads from each

You won t be able to measure the incident power precisely. The readout of the power would be lower than the real incident power.

INTRODUCTION TO MICROSCOPY. Urs Ziegler THE PROBLEM

Microscope anatomy, image formation and resolution

ABOUT RESOLUTION. pco.knowledge base

1.The Problem LIGHT-LEVEL LEVEL IMAGING. light-level level Cameras. 3. Solutions. 2. Low-light LOW-LIGHT

Aberrations and adaptive optics for biomedical microscopes

Boulevard du Temple Daguerrotype (Paris,1838) a busy street? Nyquist sampling for movement

Introduction to light microscopy

Megapixel FLIM with bh TCSPC Modules

Instant super-resolution imaging in live cells and embryos via analog image processing

ProLong Glass Antifade Mountant

High resolution extended depth of field microscopy using wavefront coding

Things to check before start-up.

FLUORESCENCE MICROSCOPY. Matyas Molnar and Dirk Pacholsky

Premium portable metrology. ModelMaker. Handheld scanners. MCAx. Articulated arms NIKON METROLOGY I VISION BEYOND PRECISION

Transcription:

Super Resolution Microscope N-SIM E Super Resolution Microscope

Explore Nano world with Nikon N-SIM E is a streamlined, affordable superresolution system that provides double the resolution of conventional optical microscopes: the same level of superb resolution as Nikon Super Resolution Microscope N-SIM. Configured with A1+ confocal microscope Simple imaging method switching The N-SIM E can be simultaneously configured with a confocal microscope system, such as the A1+ or C2+, and imaging methods can be easily switched between super resolution imaging and confocal imaging. A desired location for the SIM image can be specified in a confocal image and acquired in super-resolution. Confocal image Super-resolution image Select the location to acquire a SIM image in a confocal image Acquire the SIM image of the selected location Dry objective compatibility Dry objectives are now compatible with N-SIM E as well as confocal microscopes, so both confocal imaging and super resolution imaging are available without switching lenses. Low-magnification and wide field-of-view dry lenses enable high resolution observation even at the periphery of sample tissue. FOV of 40X dry lens FOV of 100X lens CFI Plan Apochromat Lambda 60XC CFI Plan Apochromat Lambda 40XC

The principle of the Structured Illumination Microscopy Analytical processing of recorded moiré patterns, produced by overlaying a known high spatial frequency pattern, mathematically restores the sub-resolution structure of a specimen. Utilization of high spatial frequency laser interference to illuminate sub-resolution structures within a specimen produces moiré fringes, which are captured. These moiré fringes include modulated information of the sub-resolution structure of the specimen. Through image processing, the unknown specimen information can be recovered to achieve resolution beyond the limit of conventional optical microscopes. Illumination with a known, high spatial frequency pattern allows for the extraction of super-resolution information from the resulting moiré fringes. Create super-resolution images by processing multiple moiré pattern images An image of moiré patterns captured in this process includes information of the minute structures within a specimen. Multiple phases and orientations of structured illumination are captured, and the displaced super-resolution information is extracted from moiré fringe information. This information is combined mathematically in Fourier or aperture space and then transformed back into image space, creating an image at double the conventional resolution limit. Create super-resolution images by processing multiple images Capture multiple images with structured illumination that is shifted in phase. Repeat this process for three different angles. This series of images are then processed using advanced algorithms to obtain super-resolution images. Utilizing high-frequency striped illumination to double the resolution The capture of high resolution, high spatial frequency information is limited by the Numerical Aperture (NA) of the objectives, and spatial frequencies of structure beyond the optical system aperture are excluded (Fig. A). Illuminating the specimen with high frequency structured illumination, which is multiplied by the unknown structure in the specimen beyond the classical resolution limit, brings the displaced super-resolution information within the optical system aperture (Fig. B). When this super-resolution information is then mathematically combined with the standard information captured by the objective lens, it results in resolutions equivalent to those captured with objective lenses with approximately double the NA (Fig. C). Fig. A: Resolution is limited by the NA of the objective Fig. B: The product of Structured Illumination and normally un-resolvable specimen structure produce recordable moiré fringes containing the specimen information at double the conventional resolution limit. Fig. C: Images with resolutions equivalent to those captured with objective lenses with approximately double the NA are achieved.

Objectives for super-resolution microscopes The system can be configured with either a 100X oil immersion type, which is suitable for the imaging of fixed samples, or a 60X water immersion type, which is optimal for time-lapse live-cell imaging. The SR (super resolution) objectives have been designed to provide superb optical performance with Nikon s super-resolution microscopes. The adjustment and inspection of lenses using wavefront aberration measurement have been applied to yield optical performances with the lowest possible asymmetric aberration. CFI SR HP Apochromat TIRF 100XC Oil CFI SR Plan Apochromat IR 60XC WI Auto correction collar (Option) This unique, auto correction collar with harmonic drive and automatic correction algorithm, enables perfect alignment of the correction collar of AC series objectives, easily and accurately compensating for changes in temperature, deviation of cover glass thickness, or refractive-index distribution in samples. 3-color multi-laser super-resolution capability The compact LU-N3-SIM laser unit dedicated for N-SIM E is installed with the three most commonly used wavelength lasers (488/561/640), enabling super-high resolution imaging in multiple colors. LU-N3-SIM laser unit N-SIM analysis software Image processing, reconstruction and analysis are carried out using the N-SIM E module that resides within Nikon s universal, cross-platform imaging software NIS-Elements with intuitive, simple operation. Setting image acquisition Up to five different laser wavelengths are available. User-customized spectral, z-stack, and time-lapse acquisition settings are automatically managed to allow for a simple workflow from acquisition to image reconstruction. Image reconstruction can be further optimized by modifying reconstruction parameters post-acquisition/offline. Setting image reconstruction Auto settings allow the software to automatically select the most appropriate reconstruction parameters for the acquired images. Users can further optimize reconstruction by manually adjusting these parameters. Reconstruction view Reconstruction view allows users to preview the results of the selected reconstructed parameters on the current/selected frame, allowing for efficient reconstruction parameter determination. High-speed reconstruction processing using GPU Image acquisition (3D-SIM) High-speed processing using GPU ensures image reconstruction five times faster than that of CPUs, and allows image processing with reduced stress (when using a recommended PC and GPU board). NVIDIA Quadro GPU

N-SIM E supports only essential, commonly used excitation wavelengths and imaging modes while providing the same super-resolution images as the N-SIM, making it an obvious choice for individual labs. Double the resolution of conventional optical microscopes The N-SIM E utilizes Nikon s innovative new approach to structured illumination microscopy technology. By pairing this powerful technology with Nikon s renowned CFI Apochromat TIRF series 100X Oil objective (NA 1.49), the N-SIM E nearly doubles the spatial resolution of conventional optical microscopes (to approximately 115 nm*), and enables detailed visualization of the minute intracellular structures and their interactive functions. *Excited with 488 nm laser, in 3D-SIM mode Super-resolution image (3D-SIM) Conventional widefield image Microtubules in B16 melanoma cell labeled with YFP Objective: CFI Apochromat TIRF 100X Oil (NA 1.49) Image capturing speed: approximately 1.8 sec/frame (movie) Reconstruction method: Slice Photographed with the cooperation of: Dr. Yasushi Okada, Laboratory for Cell Polarity Regulation, Quantitative Biology Center, RIKEN Super-resolution image (3D-SIM) Conventional widefield image Endoplasmic reticulum (ER) in living HeLa cell labeled with GFP Objective: CFI Apochromat TIRF 100X Oil (NA 1.49) Image capturing speed: approximately 1.5 sec/frame (movie) Reconstruction method: Slice Photographed with the cooperation of: Dr. Ikuo Wada, Institute of Biomedical Sciences, Fukushima Medical University School of Medicine Axial super-high resolution imaging with 3D-SIM mode Two reconstruction methods are available. Slice reconstruction allows axial super-resolution imaging with optical sectioning at 300 nm resolution in live-cell specimens. Optional stack reconstruction can image thicker specimens with higher contrast than slice reconstruction. Super-resolution image (3D-SIM) Conventional widefield image Bacillus subtilis bacterium stained with membrane dye Nile Red (red), and expressing the cell division protein DivIVA fused to GFP (green). The super-resolution microscope allows for accurate localization of the protein during division. Reconstruction method: Slice Photos courtesy of: Drs. Henrik Strahl and Leendert Hamoen, Centre for Bacterial Cell Biology, Newcastle University 3D-SIM (Volume view) 3D-SIM (Maximum projection) Width: 26.19 µm, Height: 27.11 µm, Depth: 3.36 µm Mouse keratinocyte labeled with an antibody against keratin intermediate filaments and stained with an Alexa Fluor 488 conjugated second antibody. Reconstruction method: Stack Photos courtesy of: Dr. Reinhard Windoffer, RWTH Aachen University Fast 1 sec/frame temporal resolution for super resolution imaging N-SIM E provides fast imaging performance for Structured Illumination techniques, with a time resolution of approximately 1 sec/frame, which is effective for live-cell imaging.

Specifications Lateral resolution (FWHM of beads in xy) 115 nm* in 3D-SIM mode Objective CFI SR HP Apochromat TIRF 100XC Oil (NA 1.49) CFI SR Apochromat TIRF 100XAC Oil (NA 1.49) CFI SR Plan Apochromat IR 60XC WI (NA 1.27) CFI SR Plan Apochromat IR 60XAC WI (NA 1.27) CFI Plan Apochromat Lambda 60XC (NA 0.95)** CFI Plan Apochromat Lambda 40XC (NA 0.95)** Axial resolution (FWHM of beads in z) 269 nm* in 3D-SIM mode Image acquisition time Up to 1 sec/frame (3D-SIM) Imaging mode 3D-SIM Reconstruction method: slice, stack (option) Camera ORCA-Flash 4.0 scmos camera (Hamamatsu Photonics K.K.) Multi-color imaging Up to 3 colors Software Compatible Laser LU-N3-SIM laser unit 488 nm, 561 nm, 640 nm Operating conditions 20 ºC to 28 ºC ( ± 0.5 ºC) Compatible microscope Motorized inverted microscope ECLIPSE Ti2-E Perfect Focus System Motorized XY stage with encoders Motorized barrier filter wheel Piezo Z stage (option) NIS-Elements Ar NIS-Elements C (for Confocal Microscope) Both require additional software modules NIS-A 6D and N-SIM Analysis Layout * These values are measured using 100nm diameter beads excited at 488nm. Actual resolution is dependent on laser wavelength and optical configuration. ** Supports slice reconstruction. System diagram 673 Ti2-E with double layer configuration with Perfect Focus Unit Motorized HG fiber illuminator Intensilight 1354 1542 TI2-LA-FL Epi-Fl module 800 Laser unit LU-N3-SIM laser unit Piezo Z stage (option) PC N-SIM Shield box Vibration isolated table N-SIM E illuminator unit PC rack NIS-Elements AR / NIS-Elements C* TI2-FTQ N-SIM motorized quad band filter turret 1000 N-SIM E NIS -A 6D and N-SIM analysis Vibration isolated table ORCA-Flash4.0 scmos camera (Hamamatsu Photonics K.K.) 1500 TI2-P-FWB-E Motorized BA filter wheel 2280 * Required when used with confocal system unit mm Specifications and equipment are subject to change without any notice or obligation on the part of the manufacturer. December 2017 2015-17 NIKON CORPORATION WARNING WARNING-LASER RADIATION AVOID EXPOSURE TO BEAM CLASS 3B LASER PRODUCT TO ENSURE CORRECT USAGE, READ THE CORRESPONDING MANUALS CAREFULLY BEFORE USING YOUR EQUIPMENT. Total Power 500mW MAX. CW 400 700nm IEC/EN60825-1 : 2007, 2014 Monitor images are simulated. Alexa Fluor and Cy are registered trademarks of Thermo Fisher Scientific, Inc. Company names and product names appearing in this brochure are their registered trademarks or trademarks. N.B. Export of the products* in this brochure is controlled under the Japanese Foreign Exchange and Foreign Trade Law. Appropriate export procedure shall be required in case of export from Japan. *Products: Hardware and its technical information (including software) NIKON CORPORATION Shinagawa Intercity Tower C, 2-15-3, Konan, Minato-ku, Tokyo 108-6290, Japan phone: +81-3-6433-3705 fax: +81-3-6433-3785 http://www.nikon.com/products/microscope-solutions/ NIKON INSTRUMENTS INC. 1300 Walt Whitman Road, Melville, N.Y. 11747-3064, U.S.A. phone: +1-631-547-8500; +1-800-52-NIKON (within the U.S.A. only) fax: +1-631-547-0306 http://www.nikoninstruments.com/ NIKON INSTRUMENTS EUROPE B.V. Tripolis 100, Burgerweeshuispad 101, 1076 ER Amsterdam, The Netherlands phone: +31-20-7099-000 fax: +31-20-7099-298 http://www.nikoninstruments.eu/ NIKON INSTRUMENTS (SHANGHAI) CO., LTD. CHINA phone: +86-21-6841-2050 fax: +86-21-6841-2060 (Beijing branch) phone: +86-10-5831-2028 fax: +86-10-5831-2026 (Guangzhou branch) phone: +86-20-3882-0550 fax: +86-20-3882-0580 Printed in Japan (1712-03)T Complies with FDA performance standards for laser products except for deviations pursuant to Laser Notice No. 50, dated June 24, 2007 ISO 14001 Certified for NIKON CORPORATION NIKON CANADA INC. CANADA phone: +1-905-602-9676 fax: +1-905-602-9953 NIKON FRANCE S.A.S. FRANCE phone: +33-1-4516-45-16 fax: +33-1-4516-45-55 NIKON GMBH GERMANY phone: +49-211-941-42-20 fax: +49-211-941-43-22 NIKON INSTRUMENTS S.p.A. ITALY phone: +39-55-300-96-01 fax: +39-55-30-09-93 NIKON GMBH SWITZERLAND SWITZERLAND phone: +41-43-277-28-67 fax: +41-43-277-28-61 NIKON UK LTD. UNITED KINGDOM phone: +44-208-247-1717 fax: +44-208-541-4584 NIKON CEE GMBH AUSTRIA phone: +43-1-972-6111 fax: +43-1-972-611-140 Code No. 2CE-SCMH-5 NIKON SINGAPORE PTE LTD SINGAPORE phone: +65-6559-3651 fax: +65-6559-3668 NIKON INSTRUMENTS KOREA CO., LTD. KOREA phone: +82-2-2186-8400 fax: +82-2-555-4415 This brochure is printed on recycled paper made from 40% used material. DANGER-VISI LASER RADI OR SIKN EXP OR SCATTER CLASS 4 LAS Total Power 1500m CW 370 790nm IEC/EN60825-1 : 2 Complies with FD for laser products deviations pursua No.50 dated June