Hi Simon / Steffan
Many thanks for this, it's very helpful. We’ve been carrying out a very similar exercise ourselves. With the CoolLed PE300 white I had I needed to swap out the excitation filter in my TRITC / RFP cube so it worked a bit better with the illumination from the Phosphor. This mitigated somewhat the effect of decreased signal for green excitation fluors.
In a side by side comparison (actually carried out by Matt Pearson and not me) we found the intensity of the LED depended rather on the filterset one was coupling the system up with. Both CoolLED and Lumencor were very happy to advise us of necessary changes in filters to make the best of what we had.
I agree that everyone should be switching, from a facility management point of view it makes life so much easier and cheaper long term and it's considerably better for live experiments and the environment.
Ann
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-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Simon Walker
Sent: 12 June 2015 14:01
To: [log in to unmask]
Subject: Re: Intensity of LED excitation (SOLA)
Hi Steffen,
I've been gradually migrating our systems to LED illuminators and have purchased several options including CoolLED pE-300, Lumencor SOLA and Lumencor Spectra-X. We now have four pE-300 units which in my opinion are ideal as a replacement for a standard mercury burner/metal halide light source on a confocal. We've used these on our Zeiss 780 and Olympus FV1000 confocals and users can distinguish no significant difference compared with the old illuminators (HXP 120 and mercury burners) in all channels. The SOLAs are in use on wide-field imaging systems where the pE-300 is no good as it doesn't have the ~630 nm LED for far red fluorophores. The only significant difference we've found here is that the DAPI excitation (~380 nm) is around 6x weaker (although as the DAPI is typically very bright this doesn't matter), and the 'Texas Red' excitation (~585 nm) is around half as bright compared with the mercury burner. All other channels are equivalent or brighter, including the 'TRITC' channel (~560 nm). The Spectra-X is a little different as although the light engine is similar (if not identical) to the SOLA, the electronics permit triggering and intensity control of individual LEDs. This works brilliantly with Nikon Elements software.
I think it's been a while coming, but in my view for most applications LED illuminators are now by far the better option. Everyone should be switching now!
Simon
P.S. No commercial interest
----------------
Simon A Walker PhD
Imaging Facility Manager
Babraham Institute
Cambridge, UK
CB22 3AT
http://www.babraham.ac.uk/science-services/imaging
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Steffen Dietzel
Sent: 11 June 2015 17:50
To: [log in to unmask]
Subject: Intensity of LED excitation (SOLA)
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Dear Listers,
we are about to fix the details on the equipment for several microscopes for a new core facility. One thing that came up was the excitation light source for conventional fluorescence for confocals.
I would very much like to switch to LEDs, therefore we consider Lumencor "SOLA-SM II - White LED source". An article refered to earlier on this list (doi: 10.7171/jbt.14-2502-001
<http://dx.doi.org/10.7171%2Fjbt.14-2502-001>) described that light intensitiy of a SOLA is higher over the whole relevant than a "120 W metal Halide" lamp (Fig 1, http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3970759/figure/F1/), so this seemed to be a good choice.
I know however heard from two sources that I consider reliable that the SOLA has significantly less output in the excitation range of orange and near red dyes (maybe 540 to 600) than a EL6000 using a HXP R 120W metal halogenid lamp.
Therefore I would appreciate if you could share your thoughts on and experience with the SOLA, in particular with orange and near red dyes.
Thanks
Steffen
--
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Steffen Dietzel, PD Dr. rer. nat
Ludwig-Maximilians-Universität München
Walter-Brendel-Zentrum für experimentelle Medizin (WBex) Head of light microscopy
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