CoolLED pE-340fura

CoolLED pE-340fura

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. 

CoolLED pE-340fura
Dedicated 340nm & 380nm outputs

Required for Fura-2 ratiometric calcium imaging

Broad white output (435nm-645nm)

Everyday imaging also possible with fluorophores including GFP/FITC, Tritc/TxRed/mCherry, Cy5 and similar

Microsecond switching (340/380)

Better temporal resolution

Stability of LED illumination

Reduction in noise level of the light source to a range below that of the wider experiment, reducing false positives and increasing overall precision. Accuracy of the Fur

Higher signal-to-noise

Gives cleaner images and data whilst requiring less Fura-2 dye, reducing toxicity and costs

UV optimized optics

Maximum power delivery on standard microscope configurations

Simple to fit and use

No alignment – once only adjustment

Direct or light guide delivery options

Flexibility of UV optimised attachment options

Individual channel intensity control (0-100%)

Offers ability to colour balance to optimise experiment set up

Individual channel selection

Control the level of excitation of each stain independently on a multi stained sample

Compatible with most imaging software

Integrated control with your imaging set up time after time

Individual channel triggering (TTL, microsecond)

Fast remote control operation

Removable inline filter holders

No moving parts

Sequence runner

Facility to run through a sequence of excitation channels using equipment with only a single TTL out

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

Images were obtained by Martin Bootman and Katja Rietdorf, School of Life, Health and Chemical Sciences, The Open University, UK.

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.

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.

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

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.

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.

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

Comparison of Ca2+ increases obtained from the application of trypsin (100 nM) to tsA-201 cells loaded with different concentrations of Fura-2 AM.

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.

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

High levels of autofluorescence and fast photobleaching of specific fluorescence when illuminating Qdots with Metal Halide.  

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.

Power requirements
Power requirements
100-240V a.c. 50/60Hz, 1.4A
Power consumption
Power consumption
Standby Max 2W
1 band (white) at 100% intensity Max 24W
2 bands (340 + white) at 100% intensity Max 30W
3 bands (All) at 100% intensity 36W
pE-340fura Light Source: 77mm(w) x 186mm(d) x 162mm(h) - Weight 1.40kg
pE-340fura Control Pod: 88mm(w) x 125mm(d) x 37mm(h) - Weight 0.32kg
pE-340fura Power Supply: 167mm(w) x 67m(d) x 35mm(h) – Weight 0.62kg
pE-Universal Collimator: 44mm(w) x 86mm(d) x 44mm(h) – Weight 0.17kg
Environment & Safety
Environment & Safety
LED products are more sustainable and energy efficient than conventional illuminators. CoolLED's products have the following benefits:
Energy Efficient: 80% less power
Long lifetime (25,000 operating hours)
No bulb replacements
Reduced risk of eye damage
Quiet operation
No special disposal regulations or issues
System = 24 months extendable by free product registration
LEDs = 36 months (NB 340nm LED warranted for 3000 hours accumulated use)

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