PE-4000

The pE-4000 is a powerful, flexible, controllable system for advanced research applications.

“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

Advanced control for cutting-edge research

Multiple levels of control are possible, from simple to advanced. A dual mode control pod gives immediate manual control in either White or Advanced modes, where individual LED channels can be individually switched on/off or irradiance modulated.

Alternatively, the USB interface and integration into major microscope software platforms allows complex software control.
TTL control enables capturing high-speed events (<1 ms). Since there is no shutter or filter wheel, without these moving parts, this reduced latency means speed is limited only by the camera. An expansion box is also available for added functionality with multiple BNC connections for remote analogue and TTL control.

For optogenetics applications, alongside the ability to include excitation filters to tune the wavelength to the opsin’s maximum activation peak, an internal function generator controls pulse timing and waveforms.

Improving cell viability

With the pE-4000, it is now possible to prevent artefacts and allow live samples to thrive in experiments >100 hours. Fine control of illumination timing and irradiance with TTL and the function generator minimises exposure, avoiding photobleaching and phototoxicity.

“When you can only control the 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 irradiance 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

Matching diverse fluorophore requirements

Full filter compatibility and 256 wavelength combinations from 16 installed LEDs (365 nm -770 nm) covers all available fluorophores, which is ideal for multi-user facilities or evolving experimental needs.

Multi-channel experiments with no pixel shift

Up to four channels using distinct wavelengths can be captured with no mechanical movement, preventing pixel shift and allowing the direct comparison of data as in the following experiment:

(A) Representative photocurrent traces of a PhobosCA expressing CA1 cell evoked with different activation wavelengths and shutoff with 595 nm light. (B) Photocurrent traces in the same cell evoked with 460 nm light and shutoff with indicated wavelengths (10 mW/mm2).1

Increased power efficiency

The pE-4000 includes 16 LEDs grouped into four individually controllable channels, and this unique Wavelength Grouping Concept makes it possible to deliver more power in an efficient four-channel system. As the first company to introduce LED microscope lighting for fluorescence microscopy, CoolLED has developed a comprehensive understanding of the complexities of configuring and selecting filter sets for experiments using multiple fluorophores. CoolLED’s innovation comes from recognising that all the fluorophores used in multi-band work can have their absorption bands divided into four separate groups across the spectrum due to the restricted bandwidth available.

DAPI/FITC/TRITC/Cy5 Quad filter excited by pE-4000 at matched wavelengths shown above.

A choice of 16 LEDs enables the user to select the optimal excitation wavelength for each fluorophore combination and optical filter configuration, which enables maximum performance and flexibility. Our Filter Finder tool provides more information on recommended filter configurations.