LEDs – More Than Just a Light Source

1. LEDs – More Than Just a Light Source Presented by Danielle Love

2. CoolLED Overview • Based one hour west of London, UK • Design, develop and manufacture specialist illumination technology • Key market is the microscopy industry • Using market knowledge to drive product development

3. Overview CoolLED Timeline Arc Lamps LEDs CoolLED Products Summary

4. Illumination Timeline 2006 Launch of pE-1 2012 Launch of pE-300 2018 Launch of enhanced pE-4000 1900’s First fluorescence microscope using transmitted light 2003 Development of first commercial LED system 2010 Launch of pE-100 2014 pE-100wht & pE-300white launched 1595 First compound microscope 2006 Introduced the first commercially available LED illumination system for microscopy 2016 Launch of pE-300lite 2011 Research into white light 1665 Robert Hook microscope with illuminator and condenser 2007 Launch of 4 colour pE-2 2013 Launch of pE-4000 1930’s Mercury lamps used for Epi-Fluorescence

5. Arc Lamps Traditionally used for fluorescence microscopy. Mercury arc lamp are most common. Many fluorophores designed around mercury peaks. High intensity, broad spectrum light source. Michael W. Davidson - National High Magnetic Field Laboratory, The Florida State University

6. Arc Lamp – Disadvantages • Bulb intensity decreases with use – can lead to misleading data. • Bulb alignment and focusing required with each new bulb. • Short life time – bulbs last approx. 200 hours. • High pressure bulb can explode – care must be taken when changing. • Uneven intensity across the spectrum – ND required due to intensity difference in between fluorophores. • Strong in UV, can be harmful to samples and user – UV filter required. • Mercury is harmful and requires careful disposal. • Inherent amplitude instability. Beacher J, Biophotonics International 2008 • Warm up period required before lamp can be used, on/off switching can shorten lamp life - shutter required. • Broad spectrum illuminates at all wavelengths - a filter wheel is required to excite at specific wavelengths.

7. LEDs Past Performance LEDs not powerful enough to illuminate some fluorophores Narrow bandwidth – don’t cover full spectrum LEDs not the correct wavelengths for fluorophores

8. LEDs Today LEDs are now powerful enough Larger number of LEDs – UV, Visible, IR Design of new fluorophores

9. Benefit 1: Improved Temporal Resolution Fast switching No mechanical shutter Improved temporal resolution Pulsing for decreased photobleaching

10. Benefit 2: Environmental Reduced energy bills – cooler rooms – no hazardous chemicals Instant on/off means that LEDs can be switched off when not in use Lower power consumption Energy efficient, emit less heat Mercury-free

11. Evans, A, Green Light Laboratories LTD, 2017

12. Benefit 3: No ND filters required Intensity electronically controlled Get the exact intensity you require Allows for colour balancing in multi colour experiments

13. Benefit 4: No changing bulbs and misleading data Very long lifetime - No need to change bulbs Factory aligned and focussed No decline in intensity

14. Benefit 5: Improved signal to noise LED illumination offers much higher amplitude stability than arc lamps. D. A. Wagenaar, PLoS One, 2012

15. Benefit 6: Filter friendly Single wavelength imaging: No need to change your existing filter set up Can use multiband filter sets

16. Benefit 6: Filter friendly Slow speed multi fluorophore imaging: Multiple LEDs in one light source No need to change existing filters. Can place exciters in front of LED Can use multiband filter sets – avoid pixel shift

17. Benefit 6: Filter friendly High speed multi fluorophore imaging: Single band sets – too slow Filter wheels – too slow/vibrations Solutions: Multi-band set or Exciters in light source with multiband dichroic: No filter wheel time delay No filter wheel vibrations Only limited by camera exposure No pixel shift

18. Benefit 6: No filter wheel

19. Where can CoolLED LEDs be used for? Transmitted illumination • Standard white light for 100W halogen replacement • Phase contrast techniques • Single wavelength for IR-DIC or specialist applications Fluorescence illumination: • Replacement for fluorescence arc lamps • Single wavelength • Multi-wavelength imaging • High speed multi-wavelength imaging

20. pT-100 – Transmitted Applications • pT-100 has 4 variants for transmitted imaging techniques including brightfield, darkfield, DIC, Dodt gradient contrast & phase contrast: • Broad white output – pT-100-WHT (formerly known as pE-100wht )a powerful white LED illumination system designed to replace a 100W halogen lamp.  Unlike a Halogen lamp, the colour balance of the pT-100-WHT Illumination System does not vary with intensity, removing the need to make any adjustment.

21. pT-100 – Transmitted Applications 2. Narrow bandwidth at 525nm – pT-100-525 is optimised for the spectral response of most scientific cameras and reduces background noise from a phosphored LED. 3. Narrow bandwidth at 635nm – pT-100-635 provides deeper penetration, excellent for revealing detail in thicker samples 4. Narrow bandwidth at 770nm – pT-100-770 provides deeper sample penetration techniques that require the use of infrared light

22. pE-100 • One LED = one channel • Select wavelength to match fluorophore • Perfect for clinical applications, optogenetics & electrophysiolgy • Manual control pod, Instant on/off, 0-100% intensity control • Fast switching by TTL trigger (<150us) • Direct-fit, LLG and Fiber options

23. pE-300Series • Three models – same spectral output, different control options • White light, broad spectrum output for fluorescence • Fits all new and most older microscopes • Replaces mercury and metal-halide lamps

24. The New pE-340fura • Bespoke LED Illuminator for Fura-2 ratiometric calcium imaging • Dedicated 340nm and 380nm outputs • Broad white output 435nm-645nm

25. pE-4000 Flexible Microscopy Illumination System pE-4000 is a powerful 4-channel high-specification LED Illumination system. It is flexible, controllable and environmentally friendly. 16 selectable available LED wavelengths, it offers the broadest spectrum of illumination available.

26. Summary Long lasting Stable Controllable Provide equivalent of: filter, attenuator, shutter and sequence generator Safer for user and environment Wide range of available wavelengths.

27. CoolLED Ltd. t +44 (0) 1264 323040 e info@coolled.com w www.coolled.com CoolLED USA t 1 800 877 0128 e info@coolled.com w www.coolled.com Thank You!