The FLUOVIEW FV3000 series is designed to meet some of the most difficult challenges in modern science. Featuring the high sensitivity and speed required for live cell and tissue imaging, the FV3000 enables 2D-6D (x,y,z,t,λ,p) macro to micro imaging of cells, tissues, and small organisms. With an intuitive and adaptable user interface, the FV3000 supports complete workflows from image acquisition to processing and analysis. Particular attention has been paid to the needs of cell biology, cancer research, and stem cell research, and with two new upright configurations, the FV3000 is also poised to meet the needs of neuroscience, electrophysiology, and developmental biology.
High Sensitivity Multi-Channel Imaging
A Fully Spectral System with Sensitivity and Accuracy
The FV3000 series employs Olympus’ TruSpectral detection technology. Based on patented* Volume Phase Hologram (VPH) transmission and an adjustable slit to control light, the spectral detection is highly efficient, enabling users to select the detection wavelength of each individual channel to 2 nm.
Efficient TruSpectral Detection System
The FV3000 is a fully spectral series of confocal microscope. TruSpectral detection delivers improved overall transmission and sensitivity. The high signal-to-noise ratio results in excellent multi-color confocal imaging capabilities.
Enhanced Quantum Efficiency
The GaAsP Photomultiplier Tubes (PMTs) in the FV3000’s high sensitivity detector (HSD) enable users to view samples whose emission is too weak to view with conventional detection methods. The GaAsP PMT unit incorporates two channels with a maximum quantum efficiency of 45%, and Peltier cooling that reduces background noise by 20% for high S/N ratio images under very low excitation light.
Multichannel TruSpectral Detection with Sixteen-Channel Unmixing
TruSpectral technology’s efficient design and software enable spectral detectors to run in multichannel mode for both live and post-processing spectral unmixing with a multichannel lambda mode. The multichannel mode facilitates constant spectral unmixing during live cell experiments, separating complex fluorescence during acquisition. With up to four different dynamic ranges from the four different channels of array, bright and dim spectral signals can be separated by independently adjusting the sensitivity of each detector.
Spectral Unmixing
The deconvolution algorithm enables overlapping spectra to be separated based on the spectral information from lambda stack images. The fluorescence cross-talk between the channels can be eliminated by the unmixing algorithm during both image acquisition and post acquisition processing.
Live Spectral Unmixing with TruSpectral Detection and Real-Time Processing
The power of TruSpectral detection plus multichannel mode means live spectral unmixing can be performed during image acquisition. Complex, overlapping spectra can be processed in real time.
Live image before unmixing of CFP (endosomes, blue), mAmetrine (plasma membrane, green), mKO (nucleus, orange) and mKeima (F-actin, purple) during time-lapse imaging. Image data courtesy of Dr. Kazuhiro Aoki, Dr. Michiyuki Matsuda, Graduate School of Medicine, Kyoto Uni
Live blind unmixed image. Image data courtesy of Dr. Kazuhiro Aoki, Dr. Michiyuki Matsuda, Graduate School of Medicine, Kyoto Un
Live 3D Rendering
See your data unfold in real time with the live 3D image display function of the FV3000 software. 3D images can be constructed during image acquisition and shown as live images.
Fucci induced Spheroid of HT29 cell line Yuji Mishima, Ph.D., Kiyohiko Hatake M.D., Ph.D. Clinical Chemotherapy, Cancer Chemotherapy Center, Japanese Foundation for Cancer Research.
Macro to Micro Imaging and Super Resolution
Macro to Micro Observation
Finding areas of interest in samples can be challenging. The confocal optical design of the FV3000 series supports macro to micro imaging from 1.25X up to 150X, so users can quickly switch from low magnification overview observation to high-magnification, detailed observation of regions of interest. Users can employ image stitching at both macro and micro levels to generate overview images that show samples in context.
A stitched image of a coronal section (30 μm thickness) from an adult YFP-H mouse cerebrum acquired with 20X objective (UPLSAPO20X). Image data courtesy of Takako Kogure and Atsushi Miyawaki, Cell Function Dynamics, Brain Science Institute of RIKEN.
Powerful One-Click Macro Analysis with cellSens
Images alone are not enough; with integrated cellSens Count and Measure analysis, the FV3000 Series can optimize images with deconvolution and analyze them with one-click macro functionality for a broad range of morphological measurements.
A spheroid image of a NMuMG cell line expressing Fucci2. Image data courtesy of Atsushi Miyawaki, Cell Function Dynamics, Brain Science Institute of RIKEN.
Olympus’ widely applicable super resolution method requires no special fluorophores and works for a wide range of samples. Ideal for colocalization analysis, the Olympus Super Resolution imaging module can acquire four fluorescent signals either sequentially or simultaneously with a resolution of approximately 120 nm*, nearly doubling the resolution of typical confocal microscopy. The imaging module is easy to use with minimal user training and can be added to any confocal system, making it a truly accessible method for achieving super resolution. * Subject to objective magnification, numerical aperture, excitation and emission wavelength, and experiment conditions.
Secondary antibody labels against GFP (Alexa Fluor 488, neurons) and SV2 (Alexa Fluor 565, red). Sample courtesy of Dr. Ed Boyden and Dr. Fei Chen, MIT.
0.5 AU Confocal Image
0.5 AU Confocal Image
Olympus Super Resolution Plus cellSens Advanced Deconvolution. Note clear separation of punctate stains with OSR.
Deconvolution
The optional constrained iterative deconvolution function improves the resolution, contrast, and dynamic ranges of confocal images obtained by the FV3000. The deconvolution function can be combined with Olympus Super Resolution (FV-OSR) to improve the z-axis resolution of the deconvolved images.
Cell line: HeLa (human cervical cancer cell line) Immunostaining: Hec1 staining (green, Alexa Fluor 488), α-tubulin staining (red, Alexa Fluor 568),DAPI staining (blue) Mitotic spindle and kinetochores are stained with anti-α-tubulin (red) and anti-Hec1 (green) antibodies, respectively. Chromosomes interact with microtubules of the mitotic spindle via kinetochores (protein structures assembled on the centromere region of chromosomes.) Image data courtesy of Masanori Ikeda and Kozo Tanaka, Department of Molecular Oncology, Institute of Development, Aging and Cancer, Tohoku University.
mage data courtesy of J. Doehner and U. Ziegler, Center for Image Analysis and Microscopy, University of Zurich
Image Analysis
The FV3000 incorporates various optional analysis functions to complete the workflow from image acquisition through data analysis. The Count and Measure solution enables the measurement of the number, size, luminosity, and morphology of the segments. Colocalization enables the analysis of overlapping fluorescent spectra.
Increase Productivity with High Speed Imaging
Galvanometer and Hybrid Galvo/Resonant Scanner
Users have their choice of two different types of scan units: galvanometer only with the FV3000 or hybrid galvanometer/resonant with the FV3000RS. The hybrid scan unit has a galvanometer scanner for high-precision scanning, as well as a resonant scanner that is ideal for high-speed imaging. With the galvanometer scanner and Olympus super resolution technology (FV-OSR), users can obtain resolutions down to 120nm with a high signal-to-noise ratio. The galvanometer scanner also features flexible scanning options, including precise tornado scanning as well as multipoint stimulation with 100ms switching time. The galvanometer scanner can image up to 16 frames per second. By switching to the resonant scanner, users can capture 30 frames per second with a full field of view at 512 x 512 pixels. By clipping down to 512 x 32 pixels, the resonant scanner can capture up to 438 frames per second to capture critical live physiological events such as calcium ion flux.
No Compromise between Speed and Field of View
Many high-speed scanning methods restrict the field of view, limiting their usefulness for examining large areas with multiple cells. The FV3000 series’ resonant scanner maintains a full 1X field of view, even at a video rate of 30 frames per second. B clipping the Y axis, additional speeds up to 438 frames per second can be achieved.
Most resonant scanners force a trade-off between speed and field of view. FLUOVIEW systems are optimized to maintain the field of view with even signal intensity so dynamic samples (e.g. calcium imaging) can be seen in the broad context of their cells and tissues. The image above shows examples of the clipped fields of view required in other resonant scanning systems.
Platelets bound to a thrombosis in the blood vessel of a mouse. Images taken at 30 fps in full frame by resonant scanner with 2 CH GaAsP PMTs.
Image data courtesy of Dr. Takuya Hiratsuka, Dr. Michiyuki Matsuda, Graduate School of Biostudies, Kyoto University.
A431 cells fixed with methanol labeled with Abcam Anti-ERK1 + ERK2 antibody (Alexa Fluor 488) ab208564 and Anti-alpha Tubulin antibody (Alexa Fluor 594) ab195889 and DAPI. Sample courtesy of Abcam.
Optimized for Live Cell Imaging
Resonant scanning greatly reduces photobleaching and phototoxicity compared to standard galvanometer scans by preventing the excitation of fluorophores into triplet states that create reactive oxygen species. These features make live cell experiments more robust and reliable. The FV3000 series has complete laser intensity control from low to high range, enabling the system to use the minimum required amount of laser power on samples. The optional laser power monitor provides consistent laser power during long-term time-lapse imaging across multiple days.
Ratio Imaging and Intensity Modulated Display (IMD)
The FV3000’s ratio imaging analysis function includes an Intensity Modulated Display (IMD) function in the software that displays quantitative fluorescence ratio changes during both standard and high-speed acquisitions. This function is particularly useful for calcium and FRET imaging where a pure ratio display provides poor contrast in background areas.
sGFP1-mito reveals heterogeneity in mitochondrial thermogenesis in HeLa cells. The images of ratio (ex 405 nm/ex 488 nm) in tsGFP1-mito-expressing cells before and after CCCP treatment at 37 °C. Scale bars indicate 10 μm (whole image) and 3 μm (inset). Image data courtesy of Shigeki Kiyonaka Ph,D, Yasuo Mori Ph,D Molecular Biology Field, Department of Synthetic Chemistry and Biological Chemistry, Kyoto University.
(Left) CFP, (Right) YFP FRET
(Left) Raw CFP/YFP ratio, (Right) IMD of CFP/YFP ratio
(Left) Raw CFP/YFP ratio, (Right) IMD of CFP/YFP ratio
Cardiomyoctye Image data courtesy of Yusuke Niino and Atsushi Miyawaki, Cell Function Dynamics, Brain Science Institute of RIKEN.
Rolling Average Processing
High-speed scanning at low laser power to avoid phototoxicity often decreases the signal-to-noise ratio. With rolling average post-processing, users have the flexibility to adjust high-speed time-lapse images while maintaining the time scale and keeping the original data.
(Left) Raw 30 fps data acquired at low laser power (0.05%, 488 nm). (Right) Rolling average processing (10 frame) on 30 fps data acquired at low laser power.
Object Tracking
In time-lapse imaging, moving objects can be automatically detected, tracked, and analyzed. cellSens software’s tracking function provides a powerful and intuitive tool to quantify dynamic processes such as cell movement and division.
Bioluminescence of RA-induced differentiating cells at day 12 from Bmal1:luc stably transfected ES cells Image data courtesy of: Kazuhiro Yagita, M.D. Ph.D. Department of Physiology and Systems Bioscience, Kyoto Prefectural University of Medicine Reference: Proc Natl Acad Sci U S A. 107(8): 3846–3851（2010）
Accurate Time-lapse Imaging
Maintain Focus with Z-Drift Compensation (ZDC) System
The IX3-ZDC2 Z-drift compensator uses minimally-phototoxic infrared light (laser class 1) to identify the location of the sample plane. One-shot autofocus (AF) mode enables several focus positions to be set as desired for deeper samples, enabling efficient Z-stack acquisitions in multiposition experiments. The continuous AF mode keeps the desired plane of observation precisely in focus, avoiding focus drift due to temperature changes or the addition of reagents, making it ideal for measurements that require more stringent focusing. Furthermore, the increased optical offset enables continuous AF with plastic vessels or with dry objectives. The Z-drift compensator is also compatible with silicone objectives (in AF mode).
Stage Control for Multi-Area Time-lapse, Microplate, and Stitching
Multi-area time-lapse and stitching provide robust and accurate time-lapse data, and enable users to generate detailed overview images to see their data in context. The well navigator function provides sophisticated, intuitive controls for a wide range of cell culture vessels and custom plates.
Stable Time-Lapse Imaging with the IX83 Microscope
A Z-drive guide installed near the revolving nosepiece combines high thermal rigidity with the stability of a wraparound structure to significantly reduce the impact of heat and vibration and improve the quality of time-lapse imaging.
Hard Disk Recording
The microscope comes equipped with a hard-disk drive (HDD) recording function. The images are stored automatically in the HDD. Large volumes of data, such as those obtained from long-term time-lapse imaging, can be easily collected.
High Contrast under Bright Conditions
The umbra unit is designed specifically for fluorescence observation under bright room conditions. It efficiently blocks out room light, enhances the contrast of fluorescence, and enables clear fluorescence observation without the need for a dark room.
Reduce Complexity with the Sequence Manager
With the Sequence Manager software module, complex protocols are handled with ease and accurate timing. Multi-day time-lapse experiments are controlled with microsecond scan accuracy and millisecond sequence execution accuracy. Various protocols, such as time-lapse with different time intervals, switching between high and low magnification, and photo-stimulation between imaging by FRAP or FRET (acceptor photobleaching), can be performed.
Life Science Analysis
The cellSens Life Science Analysis module enables analysis of images from FRAP or FRET experiments. In FRAP, τ/2 and the Mobile/ Immobile fraction can be estimated by fitting the curve of luminosity change caused by fluorescence recovery after bleaching. FRET enables the measurement of FRET efficiency by acceptor photobleaching, ratio imaging, and sensitized emission.
Superior Objectives
Silicone Immersion Objectives for Live Cell Imaging Deliver High-Resolution Observation at Depth
Olympus offers four high NA silicone immersion objectives that deliver excellent performance for live cell imaging. The refractive index of silicone oil (ne≈1.40) is close to that of living tissue (ne≈1.38), enabling high-resolution observations deep inside living tissue with minimal spherical aberration caused by refractive index mismatch. Silicone oil does not dry out or harden, so there is never a need to refill oil, making it ideal for extended time-lapse observations.
Water immersion objective (Left), Silicone immersion objective (Right)
In deep tissue observation, image quality depends on keeping the refractive index of the sample and immersion medium as close to each other as possible. When working with a silicone immersion objective, the difference between the refractive index of the samples and silicone oil is minimal, thus enabling brighter fluorescence images with higher resolution for deep tissue observation.
UPLSAPO30XS: For a broader view and greater depth Magnification: 30X, NA: 1.05 (silicone oil immersion), W.D.: 0.8 mm, cover glass thickness: 0.13 – 0.19 mm, operating temperature: 23 – 37 °C
UPLSAPO40XS : For a good balance between field of view and resolution Magnification: 40X, NA: 1.25 (silicone oil immersion), W.D.: 0.3 mm, cover glass thickness: 0.13 – 0.19 mm, operating temperature: 23 – 37 °C
UPLSAPO60XS2: For 3D observations with superior resolution Magnification: 60X, NA: 1.30 (silicone oil immersion), W.D.: 0.3 mm, cover glass thickness: 0.15 – 0.19 mm, operating temperature: 23 – 37 °C
UPLSAPO100XS: For greater brightness at depth in closely defined regions Magnification: 100X, NA: 1.35 (silicone oil immersion), W.D.: 0.2 mm, cover glass thickness: 0.13 – 0.19 mm, operating temperature: 23 – 37 °C
PLAPON60XOSC2: Enhance the Reliability of Colocalization Analysis with a Low Chromatic Aberration Objective
This oil immersion objective minimizes lateral and axial chromatic aberration in the 405–650 nm spectrum. Colocalization images are acquired reliably and images are measured with superior positional accuracy. The objective also compensates for chromatic aberration through near infrared up to 850 nm, making it the ideal choice for quantitative imaging.
Low Chromatic Aberration Objective Magnification: 60X NA: 1.4 (oil immersion) W.D.: 0.12 mm Chromatic aberration compensation range: 405 – 650 nm Optical data provided for each objective.
Performance Comparison of the PLAPON60XOSC2 and the UPLSAPO60XO
Reduce Spherical Aberration
The correction collar adjusts the lens position of objectives to correct the spherical aberration caused by refractive index mismatch, resulting in the improvement of image quality, such as resolution, brightness and contrast. The correction collar is especially necessary for objectives with high NA when they are used for super resolution imaging, because they are greatly affected by spherical aberration. The remote correction collar unit is useful for easy adjustment and improvement of the image quality, and operable on all UIS2 objectives which have a correction collar.
Choose the Frame That Suits Your Application
Inverted microscope
Suitable for observing cells cultured in a vessel.
The Z-drift compensator enables time-lapse observations that remain in focus.
Maintain the environmental conditions of cultured cells by adding a stage-top or full enclosure incubator.
Upright microscope (configured for imaging)
Optimized for fixed tissue and glass slide specimens.
Motorized 7-position nosepiece and condenser enable automated transitions from low to high magnification.
Upright microscope (configured for electrophysiology)
Ample space around the objectives enables patch-clamp devices to be installed.
Add additional space by lowering the stage position for experiments that require large sample handling.
Swing and slider nosepieces are available so objectives can be easily changed without interfering with the patch-clamp set-up.
Intuitive Software
Customizable and saveable layouts make it easy to tailor the interface to your workflow and experiment needs, from basic to complex.
Layout
Start by selecting your preferred display with specific tools for basic to complex acquisition.
Acquisition Condition
Reload settings that were ideal for your last experiment to provide consistency.
Acquisition
Activate basic to complex acquisitions with live ratio, intensity modulated display, quantitative region of interest (ROI) graphing or spectral unmixing display, and data backup for added security.
Viewer
Review data as it is generated. Generate 3D and 4D views and animations to explore and share data in depth.
Analysis
Extract data from images using online or offline processing. Analytical tools include Olympus super resolution technology (FV-OSR) and powerful cellSens software with features such as deconvolution, filtering, count and measure, and one-click macros.
Designed for use in the steel, automotive, electronics, and other manufacturing industries, the GX53 microscope delivers crisp images that can be difficult to capture using conventional microscopy observation methods. When combined with OLYMPUS Stream image analysis software, the microscope streamlines the inspection process from observation to image analysis and reporting.
Fast Inspections, Advanced Functionality
Quickly observe, measure, and analyze metallurgical structures.
Advanced Analysis Tools
1. Combined observation methods produce exceptional images
2. Easily create panoramic images
3. Create all-in-focus images
4. Capture both bright and dark areas
Optimized for Material Science
1. Software designed for materials science
2. Metallurgical analysis that complies with industrial standards
Userfriendly
Even novice operators can comfortably make observations, analyze results, and create reports.
1. Easily restore microscope settings
2. User guidance helps simplify advanced analysis
3. Efficient report generation
Advanced Imaging Technology
Our proven optics and imaging technology deliver clear images and reliable results.
1. Reliable optical performance: wavefront aberration control
2. Clear images: image shading correction
3. Consistent color temperature: high-intensity white LED illumination
4. Precise measurements: auto calibration
Modular
Choose the components you need for your application.
1. Build your system your way: fully customizable system with a variety of optional components
The MX63 and MX63L microscope systems are optimized for high-quality inspections of wafers as large as 300 mm, flat panel displays, circuit boards, and other large samples. Their modular design enables you to choose the components you need to tailor the system to your application.
These ergonomic and user-friendly microscopes help increase throughput while keeping inspectors comfortable while they do their work. Combined with OLYMPUS Stream image analysis software, your entire workflow, from observation to report creation, can be simplified.
Functional
Leading-Edge Analysis Tools
The MX63 series’ versatile observation capabilities provide clear, sharp images so users can reliably detect defects in their samples. New illumination techniques and image acquisition options within OLYMPUS Stream image analysis software give users more choices for evaluating their samples and documenting their findings.
The Invisible Becomes Visible: MIX Observation and acquisition
MIX observation technology produces unique observation images by combining darkfield with another observation method, such as brightfield, fluorescence, or polarization. MIX observation enables users to view defects that are difficult to see with conventional microscopes. The circular LED illuminator used for darkfield observation has a directional darkfield function where only one quadrant is illuminated at a given time. This reduces a sample’s halation and is useful for visualizing a sample’s surface texture.
Structure on semiconductor wafer
Condenser
Easily Create Panoramic Images: Instant MIA
With multiple image alignment (MIA), users can stitch images together quickly and easily simply by moving the KY knobs on the manual stage—a motorized stage is not necessary. OLYMPUS Stream software uses pattern recognition to generate a panoramic image, giving users a wider field of view.
Instant MIA image of a coin
Create all-in-focus images: EFI
The Extended Focus Imaging (EFI) function within OLYMPUS Stream captures images of samples whose height extends beyond the depth of focus of the objective and stacks them together to create one image that is all in focus. EFI can be executed with either a manual or motorized Z-axis and creates a height map for easy structure visualization. It is also possible to construct an EFI image while offline within Stream Desktop.
Stud bump on an IC chip
Capture Both Bright and Dark Areas Using HDR
Using advanced image processing, high dynamic range (HDR) adjusts for differences in brightness within an image to reduce glare. HDR improves the visual quality of digital images thereby helping to generate professional-looking reports.
From Basic Measurement to Advanced Analysis
Measurement is essential to quality and process control and inspection. With this in mind, even the entry-level OLYMPUS Stream software package includes a full menu of interactive measurement functions, with all measurement results saved with image files for further documentation. In addition, the OLYMPUS Stream Materials Solution offers an intuitive, workflow-oriented interface for complex image analysis. At the click of a button, image analysis tasks can be executed quickly and precisely. With a significant reduction in processing time for repeated tasks, operators can concentrate on the inspection at hand.
Efficient Report Creation
Creating a report can often take longer than capturing the image and taking the measurements. OLYMPUS Stream software provides intuitive report creation to repeatedly produce smart and sophisticated reports based on pre-defined templates. Editing is simple and reports can be exported to Microsoft Word or PowerPoint software. In addition, OLYMPUS Stream software’s reporting function enables digital zooming and magnification on acquired images. Report files are a reasonable size for easier data exchange by email.
Stand-Alone Camera Option
Using a DP22 or DP27 microscope camera, the MX63 series becomes an advanced stand-alone system. The cameras can be controlled via a compact box that requires only minimal space, helping users maximize their laboratory space while still capturing clear images and making basic measurements.
Advanced Designed to Support Cleanroom Conformity
The MX63 series is designed to work in a cleanroom and has features that help minimize the risk of contaminating or damaging samples. The system has an ergonomic design that helps keep users comfortable, even during prolonged use. The MX63 series complies with international specifications and standards, including SEMI S2/S8, CE, and UL.
Optional Wafer Loader Integration ― AL120 System*
An optional wafer loader can be attached to MX63 series to safely transfer both silicon and compound semiconductor wafers from a cassette to the microscope stage without using tweezers or wands. Renowned performance and reliability enable safe, efficient front and back macro inspections while the loader helps improve productivity in the laboratory.
MX63 combined with the AL120 wafer loader (200 mm version)
Fast, Clean Inspections
The MX63 series delivers contamination-free wafer inspections. All motorized components are housed in a shielded structure, and antistatic processing is applied to the microscope frame, tubes, breath shield, and other parts. The rotation speed of the motorized nosepieces is faster and safer than manual nosepieces, decreasing the time between inspections while keeping the operator’s hands below the wafer, reducing potential contamination.
System Design Achieving Efficient Observations
The XY stage is capable of both coarse and fine stage movements thanks to the combination of a built-in clutch and the XY knobs. The stage helps make observations efficient, even for large samples, such as 300 mm wafers. The tilting observation tube’s extensive range enables operators to sit at the microscope in a comfortable posture.
System Design Achieving Efficient Observations
The XY stage is capable of both coarse and fine stage movements thanks to the combination of a built-in clutch and the XY knobs. The stage helps make observations efficient, even for large samples, such as 300 mm wafers. The tilting observation tube’s extensive range enables operators to sit at the microscope in a comfortable posture.
Accepts All Wafer Sizes
Wafer holders and glass plates
The system works with various types of 150–200 mm and 200–300 mm wafer holders and glass plates. Should the size of the wafters change on the production line, the microscope’s frame can be modified at minimal cost. With the MX63 series, different stages can be used to accommodate 75 mm, 100 mm, 125 mm, and 150 mm wafers on the inspection line.
NEW GENERATION LEDs The pE-4000 sets the standard as the universal illumination system for fluorescence microscopy. The system has 16 selectable LED wavelengths across 4 channels that can be matched to the filters and fluorophores of almost any microscope, making it the broadest spectrum of illumination available.
The CoolLED pE-4000 now benefits from our award winning sustainable Greentechnology and delivers enhanced intensity at your microscope sample plane with a significant reduction in energy consumption, and is supplied with a 3 year warranty after registration.
The NEW Enhanced pE-4000 is a powerful 4-channel high-specification LED illumination system. It is flexible, controllable and environmentally friendly. Benefits include:
SELECTABILITY
Choice of 256 wavelength combinations from 16 installed LEDs (365nm -770nm)
Choose 1 of 4 wavelengths for each 4 channels on the control pod
Unique wavelength grouping concept (see below)
Optimum wavelength to suit your experiment
Specific wavelength characterisation for optimal optogenetic control of cell responses
Wavelength Grouping Concept
As the first company to introduce LEDs into fluorescence microscopy, CoolLED has developed a comprehensive understanding of the complexities of configuring and selecting filter sets for samples which have multiple stains. These ’multi-band’ filters must have a corresponding absorption and emission band for each stain and the bands must be kept spectrally separate to ensure there is no overlap.
DAPI/FITC/TRITC/Cy5 Quad filter excited by pE-4000 at matched wavelengths shown above.
CoolLED’s innovation comes from recognizing that all the stains used in multi-band work can have their absorption bands divided into four separate groups across the spectrum due to the restricted bandwidth available. This has allowed the development of a patent-pending, wavelength-grouping concept which makes it possible to deliver more power in an efficient four channel system.
With its unique 16 selectable LED wavelengths, the pE-4000 ensures optimum compatibility with all single and multi-band filter sets. The flexibility and extensive functionality of the pE-4000 broadens the range of illumination options in core facilities.
The pE-4000 allows multiple experiments with varying wavelengths on the same sample , as shown here.
(A) Representative photocurrent traces of a PhobosCA expressing CA1 cell evoked with different activation wavelengths and shutoff with 595nm light
(B) Photocurrent traces in the same cell evoked with 460nm light and shutoff with indicated wavelengths (10mW/mm2)
Wietek et al (2017) Anion-conducting channelrhodopsins with tuned spectra and modified kinetics engineered for optogenetic manipulation of behaviour, Scientific Reports volume 7, Article number 14957(2017) doi: 10.1038/s41598-017-14330-y
SUSTAINABILITY
Lowest Power Consumption – other LED technology uses 120-350 Watts
>25000 hours operation
Highly stable, repeatable LED technology
Market leading energy efficiency
CELL VIABILITY
Optimal illumination through control features to extend fluorescence of cells
Reduced photobleaching and phototoxicity
Microsecond switching
Variable pulse duration on/off
“When you can only control intensity of ‘white’ light (rather than individual channels), the level of photobleaching can be high. With the pE-4000, we can control the excitation of the individual channels. It is possible to optimise the excitation intensity according to the labelling, greatly reducing photobleaching and phototoxicity in a live experiment.” Dr Yan Gu, University of Sussex
“Suddenly, we were able to offer users uninterrupted extended live cell experiments of 100+ hours, without worrying about brightness fluctuations, lamps burning, room heating, etc. Also, users have reported markedly reduced bleaching and phototoxicity in their samples, both from the prokaryotic and the eukaryotic research fields.” Dr Jens Eriksson, Oslo University Hospital
CONTROLLABILITY
Advance integration into major microscope software platforms
Save wavelength combinations and return to previous set-up
USB interface
High speed TTL Triggering
Analogue control – dynamic intensity sinusoidal control option
White light control
Expansion Box
Individual channel intensity control
Pre-determined intensity saving option
VISIBILITY
High signal to noise ratio which gives cleaner images and data
“Striking, bright fluorescence images with a strong signal to noise ratio even when using a low magnification objective”
Graham Wright, Institute of Medical Biology, A*STAR Singapore
Time series of a confluent sheet of keratinocytes migrating during a live-cell imaging outgrowth assay, acquired on an Olympus IX-83 microscope equipped with a CoolLED pE-4000 lightsource (Fig. 4). Scale bar = 150 μm
ADAPTABILITY
Fits to all major research and clinical microscopes
Delivers light through a liquid light guide or fiber
Offers inline removable excitation filter holders
Uses the Universal Collimator
RELIABILITY
Assured through experience and QA assured processes
3 year Warranty after registration
First year Warranty swap
The compact pE-4000 Light Source houses all the LEDs, their combining optics, thermal management and control electronics. A dual mode control pod gives manual control in either White or Advanced modes. Light is delivered to the microscope by either a single liquid light guide (LLG) or fiber. LLGs can be used in conjunction with CoolLED’s pE-Universal Collimator which can accept a microscope adaptor from CoolLED’s extensive range.
CoolLED’s pE-300ultra is a fluorescence microscopy light source which offers intense, broad-spectrum LED illumination for imaging most common fluorescent stains. Users have access to both microsecond switching via multiple TTL inputs and the ability to mount inline excitation filters. This, when paired with today’s high performance multi band filter sets, facilitates imaging traditionally done via a white light source and a filter wheel, with all the benefits of LED.
Control pod for CoolLED pE-300 ultra & white
Manual control for instant on/off
intensity control in 1% steps from 0 – 100%
The pE-300ultra is the most controllable member of the pE-300 Series. In addition to the many features of the pE-300lite and the pE-300white, the system offers precise control over wavelength intensity and shuttering. Until now these benefits have only been accessible to users of high end, highly flexible illuminators such as the CoolLED pE-4000.
Throughout 2017 we will be releasing exciting new products and product updates. The first member of the CoolLED product range to receive the star treatment is the pE-300 Series.
As part of the award winning pE-300 Series, the NEW Enhanced pE-300ultra now delivers DOUBLE the intensity at the microscope sample plane. It allows adjustment of output in 1% steps, giving precise control. The broad spectrum covers everyday fluorophores: DAPI, CFP, Aqua, FITC, TRITC, TxRed, Cy5 and many more.
It is now possible to view and image your samples brighter than ever.
Convaleria taken with Photometrics Prime camera with 10ms exposure.
LED illumination means no mercury to dispose of and lower energy costs due to low power consumption and the precise control offered. With the new enhanced pE-300ultra, all 3 channels at full power use just 46 Watts. That is almost a quadruple efficiency increase!
This compares to other LED technology that uses 120 to 350 Watts. This leap forward in technology makes LEDs more attractive when compared to old Mercury or Metal Halide technology. Where “green” funding is available, the reduction in institutional energy usage improves the return on investment.
CoolLED’s pE-300 Series was the 2017 winner of the “Go Beyond Award” in the “Products” category for “Excellence in sustainability in laboratory and other high technology facilities” The International Institute for Sustainable Laboratories (I2SL) runs a unique awards program honouring organisations, individuals, products,and projects that are advancing sustainable, high-performance facilities.
The pE-300ultra offers triggering from multiple TTL inputs which, coupled with the ability to mount inline excitation filters, provides microsecond switching of pre-filtered excitation light. This, when paired with today’s high performance multi-band filter sets, facilitates imaging traditionally done using a white light source and a filter wheel, with all the benefits of LED, and most excitingly at speeds not previously so affordable.
The system comprises a pE-300ultra Light Source, Control Pod, set of three Excitation Filter Holders and Power Supply. It can be specified with a microscope adaptor for direct fit configuration or with a 3mm diameter liquid light guide if there is a need to keep the source of illumination remote from the microscope. An optional pE-Universal Collimator and microscope adaptor can be selected for use with a liquid light guide.
A benefit of the pE-300ultra is that stains can be viewed either individually or in combination, without filter cube changes. This makes it ideal for use with multi-band filter sets as the screening process can be simplified when fewer filter cubes are used. Independent control of the three LED channels means that the user can control the level of excitation of each fluorescent stain independently on a multi-stained sample, potentially removing the need for single band filter sets altogether. For further information on improved multi-stain testing, go to the Downloads tab.
An important additional feature of the pE-300ultra is that it includes CoolLED’s “Sequence Runner” multiple channel excitation mode. Users can define the order of their fluorophore capture using their pE-300ultra Control Pod, then the pE-300ultra light source can accept a single TTL output from the experiment set-up’s camera to initiate the step-through of a sequence of excitation channels. This feature is independent of the individual channel TTL inputs on the light source. This offers users the facility to run through a sequence of excitation channels using a camera which has only a single TTL-out.
With an expected lifetime in excess of 25,000 on hours and a comprehensive range of microscope adaptors, the pE-300ultra can be fitted to most current and older microscopes and operate for many years without aligning or replacing bulbs. The result is a safe, convenient illumination system without any additional operating costs.
There are two pE-300ultra configuration options:
Direct-fit for connecting to a microscopes – by selecting from a range of microscope adaptors which covers all current and most older models. A simple once only adjustment will allow optimisation to the optical path of the microscope.
Liquid Light Guidewith a fixed 3mm diameter, liquid light guide. An optional pE-Universal Collimator can be specified in conjunction with a microscope adaptor if required.
The pE-300ultra offers:
Clean bright illumination across the spectrum – excites common fluorescent stains
Specify for existing single and multi-band filter sets – no need to buy new filters
Instant on/off – No shutters required, no warm up or cool down
Simple to fit, simple to use – no alignment, a once only adjustment
Stable & repeatable – reliable and consistent results
Wide range of microscope adaptors – fits most microscopes
Precise intensity control in 1% steps (0-100%) – no ND filters required
Excellent uniformity over field of view – fixed and stable, no alignment necessary
Long lifetime – expected to exceed 25,000 hours of operating time
Removable inline excitation filter holders – no moving parts
Individual channel triggering via TTL in microseconds
Sequence Runner – sequenced excitation from a single TTL-out
Alexa Fluor 594 dye is a bright, red-fluorescent dye that can be excited using the 561 nm or 594 nm laser lines. For stable signal generation in imaging and flow cytometry, Alexa Fluor 594 dye is pH-insensitive over a wide molar range.
Cyan Fluorescent Protein (CFP) is a versatile biological marker for monitoring physiological processes, visualizing protein localization, and detecting transgenic expression in vivo. CFP can be excited by the 405 nm laser line and is optimally detected at 485 nm.
Cy5 dye is a bright, far-red-fluorescent dye that can be excited using the 633 or 647 nm laser lines, and visualized Cy5 filter sets. In addition to immunocytochemistry applications, Cy5 is also commonly used to label nucleic acids.
A variety of cyanine 5.5 (Cy5.5) dyes has been used to label biological molecules for fluorescence imaging and other fluorescence-based biochemical analysis. They are widely used for labeling peptides, proteins and oligos etc. Cy5.5 dyes are one type of the most common red fluorophores.
Cy7 is a near-IR fluor that is invisible to the naked eye (Excitation/emission maximum 750/776 nm). It is used in in vivo imaging applications, as well as the Cy7.5 dye.
DAPI (4′,6-diamidino-2-phenylindole) is a fluorescent stain that binds strongly to A-T rich regions in DNA. It is used extensively in fluorescence microscopy. As DAPI can pass through an intact cell membrane, it can be used to stain both live and fixed cells, though it passes through the membrane less efficiently in live cells and therefore the effectiveness of the stain is lower.
Fluorescein isothiocyanate (FITC) is a derivative of fluorescein used in wide-ranging applications including flow cytometry. FITC is the original fluoresce in molecule functionalized with an isothiocyanate reactive group (-N=C=S), replacing a hydrogen atom on the bottom ring of the structure. This derivative is reactive towards nucleophiles including amine and sulfhydryl groups on proteins.
The green fluorescent protein (GFP) is a protein composed of 238 amino acid residues (26.9 kDa) that exhibits bright green fluorescence when exposed to light in the blue to ultraviolet range. Although many other marine organisms have similar green fluorescent proteins, GFP traditionally refers to the protein first isolated from the jellyfish Aequorea victoria. The GFP from A. victoria has a major excitation peak at a wavelength of 395 nm and a minor one at 475 nm. Its emission peak is at 509 nm, which is in the lower green portion of the visible spectrum.
tdTomato is a genetic fusion of two copies of the dTomato gene (2) which was specifi cally designed for low aggregation (1). Its tandem dimer structure plays an important role in the exceptional brightness of tdTomato (Table I). Its excitation and emission maxima occur at 554 nm and 581 nm, respectively (1). Because tdTomato forms an intramolecular dimer, it behaves like a monomer, and has been used successfully for N- and C- terminal fusions. It shows excellent photostability and its half-time (t0.5) for maturation is one hour at 37°C.
Tetramethylrhodamine (TRITC) is a bright orange-fluorescent dye with excitation ideally suited to the 532 nm laser line. It is commonly conjugated to antibodies and proteins for cellular imaging applications.
Mercury : 형광 현미경법에 가장 흔히 사용하는 광원으로, 자외선 및 가시광선 영역에서 아주 밝다.
Spectrum of mercury
Xenon : 가시광선에서 상대적으로 연속적인 스펙트럼을 가진다. 염료 및/또는 시료의 스펙트럼 특성을 정량적으로 분석하는 시스템에서는 제논 램프를 사용하는 것이 좋지만, 같은 와트수의 수은 램프만큼 밝지는 않다.
Spectrum of Xenon
LED : 약 10 ~ 40nm 범위의 상대적으로 좁은 스펙트럼 대역폭 내에서 빛을 방출한다는 고유의 특징이 있다. 일반적으로 정격 수명이 10,000시간 이상이고 50,000시간 이상 사용하는 경우도 많고, 전체 정격 수명에 걸쳐 출력 강도가 비교적 일정하게 유지된다.
Spectrum of LED
Laser : 레이저
보통은 단색광의 좁은 스펙트럼 대역에서 극히 밝다. 하지만 레이저광의 간섭성으로 인해 샘플 영역에 균일한 조도로 비추기 어렵지만, Confocal 현미경에서는 갈바노 미러를 이용한 스캔 방식을 이용하여 이미지 획득시에 사용되어진다(LSCM). Spectrum of Laser
Metal-halide :
유효수명이 최대 2000시간으로, 일반적인 수은 램프의 200시간이나 제논 램프의 400시간과 비교하면 훨씬 길다.
