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
Utilising the successful pE-300 Series platform, the pE-340fura is a bespoke LED Illuminator for Fura-2 ratiometric calcium imaging, which also supports everyday fluorescence microscopy in a compact and affordable package.
The 340nm and 380nm LED illumination system provides the optimum excitation wavelengths for Fura-2-based calcium imaging allowing high-precision, stable, high-throughput imaging with video-rate time resolution.
340nm Excitation
380nm Excitation
The images above show a field of cardiac myocytes (heart cells). The cells were loaded with Fura-2 using standard conditions (i.e. incubation with 2 micromolar Fura-2 acetoxymethyl ester for 30 minutes, followed by an additional 30 minutes for de-esterification.
Images were obtained by Martin Bootman and Katja Rietdorf, School of Life, Health and Chemical Sciences, The Open University, UK.
Until recently, the response time of illumination systems for Fura-2 imaging have been limited to milliseconds due to mechanical switching of the wavelengths in arc lamp and monochromator systems. However, the new pE-340fura can be controlled via convenient BNC TTL connections for precise illumination control in as little as 20 microseconds.
Spontaneous Ca2+ events are induced in Mg2+-free HBS. (A) Representative trace from a single hippocampal neuron of Mg2+-free induced Ca2+ events imaged at 0.5 Hz and (B) representative trace from two hippocampal neurons of Mg2+-free induced Ca2+ events imaged at 24.39 Hz.
TINNING, P. W., FRANSSEN, A. J. P.M., HRIDI, S. U., BUSHELL, T. J. and MCCONNELL, G. (2017), A 340/380 nm light-emitting diode illuminator for Fura-2 AM ratiometric Ca2+ imaging of live cells with better than 5 nM precision. Journal of Microscopy. doi:10.1111/jmi.12616
Using the new pE-340fura LED Illumination System, less Fura-2 dye can be loaded into the cells whilst still maintaining the same measured calcium concentration and good signal-to-noise ratio. The reduction in required dye not only improves cell-viability due to reduced dye toxicity, but also results in a cost reduction per experiment.
Comparison of Ca2+ increases obtained from the application of trypsin (100 nM) to tsA-201 cells loaded with different concentrations of Fura-2 AM.
TINNING, P. W., FRANSSEN, A. J. P.M., HRIDI, S. U., BUSHELL, T. J. and MCCONNELL, G. (2017), A 340/380 nm light-emitting diode illuminator for Fura-2 AM ratiometric Ca2+ imaging of live cells with better than 5 nM precision. Journal of Microscopy. doi:10.1111/jmi.12616
Work by Sandrine Prost et al., from the University of Edinburgh, has shown that with independent wavelength controllable LED sources, signal-to-noise is dramatically improved over bulb systems and even over some available white wide spectrum LED sources.
High levels of autofluorescence and fast photobleaching of specific fluorescence when illuminating Qdots with Metal Halide.
Prost S et al (2016) Choice of Illumination System & Fluorophore for Multiplex Immunofluorescence on FFPE Tissue Sections. PLoS ONE 11(9): e0162419. doi:10.1371/journal.pone.0162419
There are two pE-340fura 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 Guide with a fixed 3mm diameter, liquid light guide. An optional pE-340fura Universal Collimator can be specified in conjunction with a microscope adaptor if required, containing optics optimised to transmit the 340nm.
향상된 이미지 품질과 인체공학 설계로, Olympus CKX53은 라이브 셀 관찰, 세포 샘플링 및 처리, 이미지 캡처, 그리고 형광 관찰을 포함한 다양한 세포 배양 샘플에 뛰어난 성능과 효율적인 관찰 흐름을 제공합니다.
라이브 셀 관찰
통합 위상차( iPC : Integrated Phase Contrast ) 현미경
CKX53 iPC 시스템을 이용하여 대물렌즈 배율( 4x, 10x, 20x , 40x ) 변경시, 콘덴서 측의 위상차 링슬릿의 연동 변경이 필요 없게 되어 효율적인 관찰 작업이 가능하고, 또한 위상차 설정이 틀어짐에 대한 수시 조정이 필요없어 언제나 선명한 샘플 관찰이 가능합니다.
2X 배율, FN 22의 대물렌즈로 선명하고 넓은 시야
PLN2X 대물렌즈를 위한 링 슬릿, CKX3-SLPAS는 직경 11 ㎜ , 시야수 22 ㎜ 를 갖습니다.
2X 대물렌즈는 다른 대물렌즈 보다 확연히 높은 contrast를 제공하여, 투명한 샘플도 명확하게 식별할 수 있습니다. 예를 들어, 96-웰 마이크로 플레이트 관찰시, 넓은 시야로 인하여 스테이지를 움직이지 않고 웰의 모든 세포를 관찰 할 수 있습니다.
IVC (Inversion Contrast) 기술을 사용한 3D 셀 관찰
새로 개발된 IVC 기술로, 위상차보다 시야 심도는 좁아지며, 개체의 모양이나 투명도와 관계없이 삼차원 이미지를 선명하게 합니다. 또한, IVC 관찰은 후광 효과나, 방향성 있는 그림자를 배제하여, 개체의 선명한 관찰을 가능하게 합니다. * 10X 대물렌즈 (PLCN10X, CACHN10XIPC)는 새로운 IVC 관찰에 사용할 수 있습니다.
Glass Heater for microscope
TPi-CKX53X ( Thermo Glass Plate )
Microscope:Olympus CKX53 series
Applicable stage: XY mechanical stage CKX3-MVR
Setting range: ambient ~ 60℃
Plate dimension: W190 x D138㎜
Heating area: W174 × D127㎜
Glass thickness: 0.5 ㎜
형광 관찰 (Fluorescence Microscopy)
다양한 형광 시약과 선명한 시야
100 W 수은 램프 (U-LH100HG), 130 W 고압 수은 램프 (U-HGLGPS), 그리고 타사(3rd Party) LEDs*와 같은 여러 통합 광원을 이용하여 형광 이미지를 선명하게 관찰할 수 있습니다. 일반 연구용 형광현미경 IX3 및 BX3 에서 사용하는 미러 유닛을 동일하게 사용 할 수 있습니다.
3개의 형광미러 유닛을 장착할 수 있으나, Bright Field 관찰과 위상차 관찰에 영향을 줄 수 있으니 유닛 선택시 고려할 필요가 있습니다.
Image taken by AcquCAM 23GR2 with LUCPlanFLN40x Ph2, 1x Adapter, CKX53
밝은 조건에서 높은 Contrast
“Umbra Shield”는 특히 CKX53을 사용한 형광 관찰을 위해 설계되었습니다. 차단막은 실내 광원을 효과적으로 차단하여 형광의 대비를 향상하여 밝은 실험실 조건에서도 선명한 형광 관찰이 가능합니다. 위상차를 사용하는 경우, Umbra 차단막을 들어 올려 표본에 빛을 통과시킬 수 있습니다.
CKX53 현미경은 UV 차단 코팅 덕분에 UV 살균 공정 중에 그대로 둘 수 있습니다. 이 시스템은 약 7kg (15.4lb)으로 이전 모델보다 가볍고 설치 공간이 더 작기 때문에 실험실 공간을 덜 차지합니다. 또한, 한 손으로 현미경을 움직일 수 있으며 관찰 경통의 목 부분을 이용하여 쉽게 운반 할 수 있습니다.
멸균 벤치 환경에서 간편한 세포 샘플링
CKX53의 접안렌즈와 광축/포커스 노브 사이의 거리가 짧으므로 작업자의 손의 위치를 자연스럽게 잡을 수 있어서 초점 및 셀 샘플링이 용이합니다.
다양한 세포 배양 용기를 사용할 수 있습니다.
CKX53의 공용 홀더로, 디쉬, 마이크로플레이트, 플라스크를 포함한 다양한 용기에서 배양된 세포를 확인하기 쉽습니다. 옵션 홀더가 부착되면, 최대 세 개의 35㎜ 배양 용기를 스테이지에 장착할 수 있습니다. 또한, 다양한 마이크로플레이트를 별도의 홀더 없이 다룰 수 있습니다.
다층 조직 플라스크(Multi-Layer Tissue Flask)를 위한 종합적인 관찰
CKX53의 폭과 탈착 가능한 콘덴서로 다층 조직 플라스크와 같은, 최대 190 ㎜ 높이의 배양 용기도 볼 수 있습니다. PLCN4X 대물렌즈의 우수한 초점 심도로 다층 조직 플라스크 내 바닥 두 개의 층의 세포를 빠르고 편하게 관찰할 수 있습니다.
다양한 용기를 사용하여 관찰 유연성 증대
홀더 암을 들어 올려서 수동으로 세포 배양 용기를 배치할 수 있습니다. 또한, 스테이지는 좌우로 최대 70 ㎜ 까지 확장할 수 있습니다.
