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.
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 | 100-240V a.c. 50/60Hz, 1.4A |
|---|---|
| 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 | |
| Dimensions | 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 | LED products are more sustainable and energy efficient than conventional illuminators. CoolLED's products have the following benefits: |
| Mercury-free | |
| 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 | |
| Warranty | System = 24 months extendable by free product registration |
| LEDs = 36 months (NB 340nm LED warranted for 3000 hours accumulated use) |