pE-4000

• From my perspective the pE-4000 has improved our system in many ways; it is one of the best upgrades we have done recently, and I would never go back.

  Dr Christian Rudolph, Principal Investigator/Reader, Brunel University London

• The large range of wavelengths available in the CoolLED pE-4000 and the ability to digitally trigger them with millisecond precision when stimulating neurons allows greater flexibility in choosing the most appropriate opsin for a given experiment and stimulating it at the peak of its excitation spectrum.

  Alessandro Galloni, PhD Student, The Rancz Lab, Francis Crick Institute, London

• The CoolLED pE-4000 device performed excellently in our electrophysiological experiments and provided us with a convenient way to investigate the integrative properties of hippocampal neurons in great detail.

  Attila Szücs, Eötvös Lóránd University, Hungary

• Our CoolLED pE-4000 is installed on the spinning disk confocal. Our use of the pE-4000, for now, has been mostly limited to specimen finding and focusing. We can certainly already say that it is a very powerful and very flexible system compared to the traditional fluorescent lamp that we've installed on the other microscopes. The fact of not having neutral density filters makes the variation in light intensity much more linear. To date, we have verified that for many samples it is sufficient to use LEDs with a maximum power of 5-10%. We are also very confident about the maximum lifespan of LEDs compared to that of classic fluorescent lamps. The system interface is very user friendly, both through the Nikon NIS software to its controller.

  Professor Alex Costa, Professor in Plant Physiology, Univeristà degli Studi di Milano, Italy

• Our precisExcite is working in a fantastic manner for over 10 years. It has already contributed to more than 10 publications from the lab and will continue to do so.

  Charles Harata, Associate Professor, University of Iowa

• I have been using a pE-4000 for optogenetic experiments using in vitro slices for just over a year now. The pE-4000 has been fantastic for my needs: the ability to simultaneously use multiple wavelengths of light has allowed me to carry out experiments using two different opsins to study how external inputs converge on hippocampal interneurons. I’m currently setting up my own research group at the University of Exeter, and all of my patch-clamp rigs will have a pE-4000 on them, as they are such versatile devices for optogenetic experiments.

  Dr Michael Craig, Research Fellow, University of Exeter Medical School

• Our group is currently attempting to isolate and stain circulating tumour cells. In order to identify these cells we need a minimum of four colour fluorescence imaging, requiring the slide to be scanned using four different LEDs. To stitch our images together after scanning we require a transmission light microscope image. The image co-ordinates of the light microscope image are used to stitch the fluorescence images together. The previous light source (pE-2) allowed only three LEDs to be used together for our purposes. This meant that when we have multiple slides there are two options to get the full four fluorescence scan:1) Change the LED after scanning in Brightfield and three fluorescence channels – this risks damage to the LED itself and is time consuming. It involves un-screwing a panel each time and screws can be damaged or lost relatively easily. It places demand on the ribbon that is connected to the LED and, over time, may damage the ribbon also. We usually scan multiple slides and prefer to use option two, to reduce wear and tear on the light source.2) This option is to scan all slides in Brightfield and three colour fluorescence and then go for a second batch of images in both Brightfield and the final fluorescence channel. This is also time consuming, as setting up each scan in focus takes time. It also requires a lot more downstream processing to line the images up with one another, as the scans are often of two different sizes.  With the new pE-4000 light source I was able to scan four slides in Brightfield and four fluorescence channels in three hours. Four slides took six hours to scan on the previous light source, due to double the number of scans and changing the LED. It has also reduced the downstream processing of the images, as they are all taken using the same co-ordinates, in one batch.  Our project aims to expand from four colour fluorescence to six or seven channels over the coming years. Using the old light sources this would require a minimum of three but possibly four scans taken from different positions per slide and would take hours per slide. The current light source meets the needs of researches using modern immunofluorescence techniques as most projects have moved from single to a multi-marker approach, in recent years.

  Anthony Cooney, PhD Student, Department of Histopathology, Trinity College Dublin.

• I have been imaging using the new CoolLED pE-4000 system on our epi-fluorescence microscope. This is an excellent system as it covers a very wide spectrum of wavelengths. I have found it very easy to use. The user interface is much better as the different LEDs can be changed using the handheld control- without switching off and unscrewing the LAMs as had to be done on the old pE-2 system. In addition the LEDs can be changed dynamically while the software to capture the image is open, and also a huge combination of different wavelengths can be used at the same time. I have found that the light source from this system is considerably stronger than from the previous system - I now image at 50% strength, when I used to image at 100%. I anticipate using this imaging system a lot more now that the new LED light source is there.

  Alison Reynolds, PhD Research Fellow, UCD Conway Institute, University College Dublin

• I am very impressed with the brightness and performance of the CoolLED pE-4000 in comparison to the mercury or halide light sources for fluorescence. I also really like its neat packaging and it is easy to set up and configure. Absolutely brilliant upgrade for archaic mercury set ups!

  Dominic White, Area Sales Manager, Carl Zeiss Microscopy GmbH

• Throwing out the old mercury lamp and exchanging it for the pE-4000 on one of our live cell wide field microscopes has been an astounding success! It allowed us to cheaply breathe new life into old equipment. 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. As a facility manager I am pleased with the instrument's ease of use, and shallow learning curve for new users. Taken together with reduced overall microscope maintenance, the pE-4000 has already saved me a substantial amount of time in my daily work life, and improved the quality of our services at the same time.

  Dr. Jens Eriksson, Manager, Superresolution Microscopy Core Facility, Dep. Clinical Biochemistry, Oslo University Hospital.

• I just purchased a pE-4000 to replace an old version of Lumencor Sola SE on a live imaging microscope. After comparing the power level of the systems, I am convinced that the pE-4000 provides stronger illumination at the objective. As the Sola light source can only control intensity of ‘white’ light (rather than individual channels), we have to choose the excitation level according to the dimmest labelling fluorophore. The other channels are in turn exposed to an unnecessary high dose of excitation. The level of photo-bleaching can be high as a result. With the pE-4000, we can control the excitation of the individual channel. It is possible to optimise the excitation intensity according to the labelling, greatly reducing photo-bleaching and photo-toxicity in a live experiment. Due to the difference in the excitation spectrum with the Sola, I was worried about the extra cost of changing the dichroic, excitation and emission filters. The options of four wavelength in each channel (totally four channels) enables me to pick up the correct wavelength according to the old filter/dichroic settings.

  Dr. Yan Gu, Manager, Analytical and Quantitative Light Microscopy Facility, Genome Damage and Stability Centre, University of Sussex

Research Papers

• In vivo spatiotemporal control of voltage-gated ion channels by using photoactivatable peptidic toxins
• Two opposing hippocampus to prefrontal cortex pathways for the control of approach and avoidance behaviour
• Low membrane fluidity triggers lipid phase separation and protein segregation in living bacteria
• Organic anion transporter 1 is an HDAC4-regulated mediator of nociceptive hypersensitivity in mice
• Magnetic resonance imaging of cerebrospinal fluid outflow after low-rate lateral ventricle infusion in mice

White Papers

• Minimising photodamage during fluorescence microscopy with LED illumination
• Optimising irradiance for fluorescence microscopy
• Measuring illumination intensity with accuracy and precision

• LEDs Illuminate the Strathclyde Mesoscope
• Epifluorescence Imaging and Optogenetic Stimulation for In-vitro Electrophysiology
• CoolLED pE-4000 working with cultured hippocampal neurons
• Applying microscopy to understand skin fragility disorders

• CoolLED pE-4000 Leaflet
• CoolLED pE-4000 Quick Start Guide
• CoolLED pE-4000 User Manual
• CoolLED pE-4000 Accessories
• CoolLED pE-4000 Wavelength Grouping Concept
• CoolLED pE-4000 Troubleshooting Guide
• CoolLED pE-4000 Exploded Diagram
• CoolLED pE-4000 Declaration of Conformity