LEDs

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Dirk Dirk
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LEDs

Hi,

I was wondering if anybody has some experience with using LEDs as  
excitation sources for fluorescence microscopy and could recommend me  
some companies and/or specific models.

We are interested in equipping a Nikon Ti with LEDs instead of the  
conventional mercury lamp (e.g. Nikon's Intensilight). The main  
application of the microscope would be quantitative high-throughput  
time-lapse microscopy.

Thanks a lot. Dirk
James Pawley James Pawley
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Re: LEDs

>Hi,
>
>I was wondering if anybody has some experience with using LEDs as
>excitation sources for fluorescence microscopy and could recommend
>me some companies and/or specific models.
>
>We are interested in equipping a Nikon Ti with LEDs instead of the
>conventional mercury lamp (e.g. Nikon's Intensilight). The main
>application of the microscope would be quantitative high-throughput
>time-lapse microscopy.
>
>Thanks a lot. Dirk


A good place to start might be Chapter 6 of The Handbook of
Biological Confocal Microscopy.

Cheers,

Jim P.

--
               **********************************************
Prof. James B. Pawley,                          Ph.  608-263-3147
Room 223, Zoology Research Building,              
FAX  608-265-5315
1117 Johnson Ave., Madison, WI, 53706  
[hidden email]
3D Microscopy of Living Cells Course, June 12-24, 2010, UBC, Vancouver Canada
Info: http://www.3dcourse.ubc.ca/             Applications due by March 15, 2010
               "If it ain't diffraction, it must be statistics." Anon.
Craig Brideau Craig Brideau
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Re: LEDs

In reply to this post by Dirk
I bought a high-powered LED meant for architectural lighting (normally
put in a spotlight outside a building) and duct-taped it (literally)
onto the side of an old microscope we found in a junk heap with no
lamp.  I put a cheap condenser lens in front of the LED array to
collimate it with pretty decent results.  The LED is actually an array
of RGBA elements (red, green, blue, amber) so I have four wavelengths
to choose from.  Control is provided by a USB connection to a PC.  It
works well enough for our cell culture work, where we just want a
quick check of whether or not a given cell stained properly.  We use
epi-fluorescene using the LEDs.  The one drawback of them is the blue
LED is centered more at 460nm, so it is not so great for some dyes
like CFP, but it works well for other dyes.  In particular, the Amber
LED is 590, which works well for m-cherry (I believe it is what they
were using).

Craig

On Sun, Jan 10, 2010 at 8:20 PM, James Pawley <[hidden email]> wrote:

>> Hi,
>>
>> I was wondering if anybody has some experience with using LEDs as
>> excitation sources for fluorescence microscopy and could recommend me some
>> companies and/or specific models.
>>
>> We are interested in equipping a Nikon Ti with LEDs instead of the
>> conventional mercury lamp (e.g. Nikon's Intensilight). The main application
>> of the microscope would be quantitative high-throughput time-lapse
>> microscopy.
>>
>> Thanks a lot. Dirk
>
>
> A good place to start might be Chapter 6 of The Handbook of Biological
> Confocal Microscopy.
>
> Cheers,
>
> Jim P.
>
> --
>              **********************************************
> Prof. James B. Pawley,                                      Ph.
>  608-263-3147 Room 223, Zoology Research Building,               FAX
>  608-265-5315
> 1117 Johnson Ave., Madison, WI, 53706  [hidden email]
> 3D Microscopy of Living Cells Course, June 12-24, 2010, UBC, Vancouver
> Canada
> Info: http://www.3dcourse.ubc.ca/            Applications due by March 15,
> 2010
>               "If it ain't diffraction, it must be statistics." Anon.
>
Alessandro Esposito Alessandro Esposito
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Re: LEDs

In reply to this post by Dirk
Dear Dirk,
    it is a while I do not screen the market, but the ones I used up to 3 years
ago were Luxeon LEDs from Philips Lumileds or LEDs by Nichia. We used them
also for fluorescence lifetime imaging microscopy.

You could have a look also to this page
http://www.plantsci.cam.ac.uk/Haseloff/imaging/cheaposcope/cheaposcope.ht
m
edited by Jim Haseloff.

Kind regards,

Alessandro
---
Dr Alessandro Esposito
Senior Investigator Scientist
MRC Cancer Cell Unit
Hutchison/MRC Research Centre

http://www.quantitative-microscopy.org
http://www.wikiscope.org

---

The information in this email is confidential and may be legally privileged. It is
intended solely for the addressee. If you receive this email by mistake, please
notify the sender and delete it immediately. Opinions expressed are those of
the individual and do not necessarily represent the opinion of Hutchison/MRC
Research Centre.
Straatman, Kees (Dr.) Straatman, Kees (Dr.)
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Re: LEDs

In reply to this post by Dirk
Dear Dirk,

Here in Leicester we have a Nikon Ti with COOLLED PrecisExcite High Power LED Fluorescent Excitation System, using 400, 490 and 565 nm LEDs and EM-CCD. The LED system was bought with the microscope via Nikon. The system is mostly used for yeast imaging and gives very good results, due to reduced photo bleaching, as compared to our Leica SP5 confocal.

Best wishes

Kees

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Dirk Landgraf
Sent: 11 January 2010 00:22
To: [hidden email]
Subject: LEDs

Hi,

I was wondering if anybody has some experience with using LEDs as  
excitation sources for fluorescence microscopy and could recommend me  
some companies and/or specific models.

We are interested in equipping a Nikon Ti with LEDs instead of the  
conventional mercury lamp (e.g. Nikon's Intensilight). The main  
application of the microscope would be quantitative high-throughput  
time-lapse microscopy.

Thanks a lot. Dirk
Csúcs  Gábor Csúcs Gábor
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Re: LEDs

In reply to this post by Dirk
Dear Dirk,

We have on several of our systems a LED source from CoolLed
(http://www.coolled.com/). They work nicely.

Greetings   Gabor

No commercial interest!
Knecht, David Knecht, David
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Re: LEDs

I have been using Thorlabs LED's for a while.  They come individually in a cheap version with driver and collimation optics and mountings for different microscopes, or an integrated 4 wavelength unit with controller.  I have built a two channel unit for my teaching lab with Micro-Manager control.  Another new company with a multiwavelength unit is Prizmatix and Zeiss makes the Colibri.  These are in rough order of cost.  Dave

On Jan 11, 2010, at 5:49 AM, Gabor Csucs wrote:

Dear Dirk,

We have on several of our systems a LED source from CoolLed
(http://www.coolled.com/). They work nicely.

Greetings   Gabor

No commercial interest!

Dr. David Knecht    
Department of Molecular and Cell Biology
Co-head Flow Cytometry and Confocal Microscopy Facility
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)


lgelman lgelman
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Re: LEDs

In reply to this post by Dirk
Hi Dirk,

We have equipped 2 wide-field systems (a Zeiss Axiovert200M and an
Olympus IX70) running with Metamorph and 2 LSM510 (just for quick
visualization of the samples) with CoolLED PrecisExcite lights. They
just work great.

Best,

Laurent.

___________________________
Laurent Gelman, PhD
Friedrich Miescher Institut
Facility for Advanced Imaging and Microscopy
WRO 1066.2.16
Maulbeerstrasse 66
CH-4058 Basel
+41 (0)79 618 73 69


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of Dirk Landgraf
Sent: lundi, 11. janvier 2010 01:22
To: [hidden email]
Subject: LEDs

Hi,

I was wondering if anybody has some experience with using LEDs as  
excitation sources for fluorescence microscopy and could recommend me  
some companies and/or specific models.

We are interested in equipping a Nikon Ti with LEDs instead of the  
conventional mercury lamp (e.g. Nikon's Intensilight). The main  
application of the microscope would be quantitative high-throughput  
time-lapse microscopy.

Thanks a lot. Dirk
Mark Rand Mark Rand
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Re: LEDs

In reply to this post by Dirk
***Commercial Response***

Hi Dirk,

The range of choices for LED fluorescence illumination is growing daily. As
already mentioned, there are DIY projects as well as 3rd-party LED
illumination accessories to add to your existing microscope. These can be
integrated with software control using appropriate drivers and software
packages, for excellent benefits as previously described.

My company manufactures an all-in-one LED fluorescence microscope that
goes a step further to provide full integration of the LED illumination system
with the optics and software control. It's designed around LED "light cubes"
located near the objectives, with a very short, efficient light path. Eliminating
the traditional epifluorescence and lamphousing attachments allows it to be
very compact; it also has a built-in computer to further reduce the footprint.
This represents the opposite end of the spectrum of LED fluorescence choices,
compared to the nice DIY projects mentioned by Dave, Craig and Allessandro. If
you prefer an "out of the box" solution, this is a possibility you might want to
explore.


Cheers,


-- Mark

**********************
Mark Rand, Ph.D.
Product Manager and Senior Applications Scientist
Advanced Microscopy Group (EVOS)
www.amgmicro.com
Craig Brideau Craig Brideau
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Re: LEDs

I got a few queries about the LED I used, so here's the scoop:

The LED is an Enfis Uno Plus:

The LED 'kit' sold at Digikey:
http://search.digikey.com/scripts/DkSearch/dksus.dll?Detail&name=722-1033-ND

The LED manufacturer:
http://www.enfis.com/products/unoplusseries/default.htm

Enfis sells a kit (first link is the kit retailed by Digikey) which
runs via USB and comes with basic control software. Basically there
are four slider controls that allows you to adjust the power of each
of the four colors (or turn any or all of them on or off).

As for mounting, take the LED, put a condenser lens in front of it
about an inch or two away (play with this to get a good beam) and
attach it all to the lamp port on any microscope.  I used Thorlabs
60mm cage parts to hold the lens (a cheap condenser lens, also from
thor labs cost about $10) and the LED array, and they also sell an
adapter to allow the cage to couple to the lamp ports on most major
brands of microscopes.  I also put a fine grit diffuser in the beam to
even out the light distribution somewhat.  The mounting holes on the
LED array will mate with two of the four holes on a standard Thor 60mm
cage plate.

Craig
jacques jacques
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Re: LEDs

In reply to this post by Dirk
Hi all,

two years later, and a quite similar question.

I use a Zeiss epifluorescence microscope, and I'm thinking to replace my HBO housing by a LED multicolor source.
Of course, Zeiss sells the colibri setup, but the choice on the market is growing day after day, and for example, Thorlabs also sells something with 4 colors and a control unit, with the Zeiss mounting system.
I'm sure many other manufacturers do the same.

Do anybody compared the HBO to a LED system for classical epifluorescence ? What should be the LED power to have something similar to a 100W at common wavelenghts (dapi, gfp, rhodamine, etc) ? Even after checking this page :
http://www.olympusfluoview.com/theory/noncoherentsources.html
It is still difficult to know which valeus and units should be considered...

Thanks for the help

J.

Jacques FATTACCIOLI

Département de Chimie
Ecole Normale Supérieure
24 rue Lhomond 75231 Paris Cedex 05
Email : jacques.fattaccioli@ens.fr
Web : http://jacquesfattaccioli.wordpress.com
Gregg Jarvis Gregg Jarvis
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Re: LEDs

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Jacques,
Be careful what you buy.
Some LED lights have out of band leaks that cause other colors to be seen
by your detector and the watts provided are a bit different in color
correction.
This might not be true for your application, but food for thought
Gregg Jarvis
Senior Spectroscopist
Omega Optical Inc.
Brattleboro, VT. 05301
[hidden email]



On Wed, Feb 22, 2012 at 5:07 AM, jacques <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi all,
>
> two years later, and a quite similar question.
>
> I use a Zeiss epifluorescence microscope, and I'm thinking to replace my
> HBO
> housing by a LED multicolor source.
> Of course, Zeiss sells the colibri setup, but the choice on the market is
> growing day after day, and for example, Thorlabs also sells something with
> 4
> colors and a control unit, with the Zeiss mounting system.
> I'm sure many other manufacturers do the same.
>
> Do anybody compared the HBO to a LED system for classical epifluorescence ?
> What should be the LED power to have something similar to a 100W at common
> wavelenghts (dapi, gfp, rhodamine, etc) ? Even after checking this page :
> http://www.olympusfluoview.com/theory/noncoherentsources.html
> It is still difficult to know which valeus and units should be
> considered...
>
> Thanks for the help
>
> J.
>
>
> -----
>
> Jacques FATTACCIOLI
>
> Département de Chimie
> Ecole Normale Supérieure
> 24 rue Lhomond 75231 Paris Cedex 05
> Email : [hidden email]
> Web : http://jacquesfattaccioli.wordpress.com
> --
> View this message in context:
> http://confocal-microscopy-list.588098.n2.nabble.com/LEDs-tp4283200p7307846.html
> Sent from the Confocal Microscopy List mailing list archive at Nabble.com.
>
Hugh Newman Hugh Newman
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Re: LEDs

In reply to this post by jacques
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****


The Sola from Lumencor is a great epifluor source from what I hear. Spectrum output is similar to that of typical metal halide systems like the x-cite, and you don't have to worry about modules or any of that stuff. It puts out continuous white light from 380-680nm, and lasts 15k hours.

Check it out here http://www.lumencor.com/products/sola_product_sheet.html

Hugh Newman
 
Graduate Researcher

Dept. Physics and Physical Oceanography

Memorial University

St. Johns, Newfoundland, Canada



> Date: Wed, 22 Feb 2012 02:07:47 -0800
> From: [hidden email]
> Subject: Re: LEDs
> To: [hidden email]
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi all,
>
> two years later, and a quite similar question.
>
> I use a Zeiss epifluorescence microscope, and I'm thinking to replace my HBO
> housing by a LED multicolor source.
> Of course, Zeiss sells the colibri setup, but the choice on the market is
> growing day after day, and for example, Thorlabs also sells something with 4
> colors and a control unit, with the Zeiss mounting system.
> I'm sure many other manufacturers do the same.
>
> Do anybody compared the HBO to a LED system for classical epifluorescence ?
> What should be the LED power to have something similar to a 100W at common
> wavelenghts (dapi, gfp, rhodamine, etc) ? Even after checking this page :
> http://www.olympusfluoview.com/theory/noncoherentsources.html
> It is still difficult to know which valeus and units should be considered...
>
> Thanks for the help
>
> J.
>
>
> -----
>
> Jacques FATTACCIOLI
>
> Département de Chimie
> Ecole Normale Supérieure
> 24 rue Lhomond 75231 Paris Cedex 05
> Email : [hidden email]
> Web : http://jacquesfattaccioli.wordpress.com
> --
> View this message in context: http://confocal-microscopy-list.588098.n2.nabble.com/LEDs-tp4283200p7307846.html
> Sent from the Confocal Microscopy List mailing list archive at Nabble.com.
     
Periasamy, Ammasi (ap3t) Periasamy, Ammasi (ap3t)
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Re: LEDs

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hello
X-Cite also has a new LED system that is close to the price of the Sola but brighter for DAPI and GFP.  You are able to control individual LEDs which I don't know whether you can do with the Sola.
X-Cite also has some useful ways to program very fast sequential pulsing of the LEDs and they guarantee 20k hours.  
Check it out here http://www.ldgi-xcite.com/products-xcite-xled1.php

Dr. Ammasi Periasamy
Professor & Center Director
Keck Center for Cellular Imaging (KCCI)
Biology, Gilmer Hall (064), 485 McCormick Rd
University of Virginia
Charlottesville, VA 22904
(Campus Mail - P.O. Box 400328)
Voice: 434-243-7602 (Office); 982-4869 (lab)
Fax:434-982-5210; Email:[hidden email]
http://www.kcci.virginia.edu/contact/peri.php
************************
11th Annual Workshop on FRET Microscopy, March 5-10, 2012
http://www.kcci.virginia.edu/workshop/workshop2012/
*************************


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Hugh Newman
Sent: Wednesday, February 22, 2012 9:35 AM
To: [hidden email]
Subject: Re: LEDs

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****


The Sola from Lumencor is a great epifluor source from what I hear. Spectrum output is similar to that of typical metal halide systems like the x-cite, and you don't have to worry about modules or any of that stuff. It puts out continuous white light from 380-680nm, and lasts 15k hours.

Check it out here http://www.lumencor.com/products/sola_product_sheet.html

Hugh Newman
 
Graduate Researcher

Dept. Physics and Physical Oceanography

Memorial University

St. Johns, Newfoundland, Canada



> Date: Wed, 22 Feb 2012 02:07:47 -0800
> From: [hidden email]
> Subject: Re: LEDs
> To: [hidden email]
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi all,
>
> two years later, and a quite similar question.
>
> I use a Zeiss epifluorescence microscope, and I'm thinking to replace
> my HBO housing by a LED multicolor source.
> Of course, Zeiss sells the colibri setup, but the choice on the market
> is growing day after day, and for example, Thorlabs also sells
> something with 4 colors and a control unit, with the Zeiss mounting system.
> I'm sure many other manufacturers do the same.
>
> Do anybody compared the HBO to a LED system for classical epifluorescence ?
> What should be the LED power to have something similar to a 100W at
> common wavelenghts (dapi, gfp, rhodamine, etc) ? Even after checking this page :
> http://www.olympusfluoview.com/theory/noncoherentsources.html
> It is still difficult to know which valeus and units should be considered...
>
> Thanks for the help
>
> J.
>
>
> -----
>
> Jacques FATTACCIOLI
>
> Département de Chimie
> Ecole Normale Supérieure
> 24 rue Lhomond 75231 Paris Cedex 05
> Email : [hidden email]
> Web : http://jacquesfattaccioli.wordpress.com
> --
> View this message in context:
> http://confocal-microscopy-list.588098.n2.nabble.com/LEDs-tp4283200p73
> 07846.html Sent from the Confocal Microscopy List mailing list archive
> at Nabble.com.
     
jacques jacques
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Re: LEDs

In reply to this post by jacques
Thanks for your help and comments !

Jacques

Jacques FATTACCIOLI

Département de Chimie
Ecole Normale Supérieure
24 rue Lhomond 75231 Paris Cedex 05
Email : jacques.fattaccioli@ens.fr
Web : http://jacquesfattaccioli.wordpress.com
Julio Vazquez Julio Vazquez
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Re: LEDs

In reply to this post by jacques
*****
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http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hello Jacques,

How much power you get at the sample will depend to some degree on the light path and light delivery system of your specific microscope, in addition to the light source itself.

The only LED system we have is the Insight SSI (solid state illuminator) on our DeltaVision system. It has LEDs rated from 22 mW (505-515 nm) to 89 mW (563-588 nm). I am assuming these are nominal LED output power levels, although the brochure does not specify. For our lines, we measured 20 to 50 mW at the sample, or about 35-50% of the rated power for any given line, which would be really good delivery efficiency (our laser scanning confocal is about 10%). This is several times the power levels we were getting with a conventional mercury bulb and excitation filters on a comparable (but not identical) system (about 6 to 20 mW depending on excitation filter used, if I remember correctly). For most wavelengths, this is as bright or brighter than our Exfo or Xenon illuminators, which are typically brighter (at the sample) than our older Hg bulb illuminators. However, these illumination systems are also installed on newer microscopes with possibly improved light delivery. We haven't had the system for long enough to encounter any potential issues, but we like the brightness and the speed of wavelength switching. Also, we do not have to worry about burned out excitation filters or UV leaking, and we do not have to worry about replacing bulbs, heat, or keeping track of when  lamps are turned on and off. Also, no worries about anyone forgetting to turn off the lamp on Friday night. I can't think of any downside at this time. My second choice in terms of convenience would be the Exfo or similar systems (long life and easy to replace bulbs, combined with good brightness).

For reference, the InsightSSI specs can be found here:

http://www.api.com/downloads/pdfs/lifescience/InsightSSI.pdf


Julio Vazquez
Fred Hutchinson Cancer Research Center
Seattle, WA

http://www.fhcrc.org

==


On Feb 22, 2012, at 2:07 AM, jacques wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi all,
>
> two years later, and a quite similar question.
>
> I use a Zeiss epifluorescence microscope, and I'm thinking to replace my HBO
> housing by a LED multicolor source.
> Of course, Zeiss sells the colibri setup, but the choice on the market is
> growing day after day, and for example, Thorlabs also sells something with 4
> colors and a control unit, with the Zeiss mounting system.
> I'm sure many other manufacturers do the same.
>
> Do anybody compared the HBO to a LED system for classical epifluorescence ?
> What should be the LED power to have something similar to a 100W at common
> wavelenghts (dapi, gfp, rhodamine, etc) ? Even after checking this page :
> http://www.olympusfluoview.com/theory/noncoherentsources.html
> It is still difficult to know which valeus and units should be considered...
>
> Thanks for the help
>
> J.
>
>
> -----
>
> Jacques FATTACCIOLI
>
> Département de Chimie
> Ecole Normale Supérieure
> 24 rue Lhomond 75231 Paris Cedex 05
> Email : [hidden email]
> Web : http://jacquesfattaccioli.wordpress.com
> --
> View this message in context: http://confocal-microscopy-list.588098.n2.nabble.com/LEDs-tp4283200p7307846.html
> Sent from the Confocal Microscopy List mailing list archive at Nabble.com.
Julio Vazquez Julio Vazquez
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Re: LEDs

In reply to this post by jacques
*****
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http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hello all,

Applied Precision just told me that the InsightSSI illuminator is actually not LED based, so please ignore my previous posting, as far as LEDs are concerned.

Julio.

==


On Feb 23, 2012, at 2:06 AM, jacques wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Thanks for your help and comments !
>
> Jacques
James Pawley James Pawley
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Re: LEDs

In reply to this post by Julio Vazquez
*****
To join, leave or search the confocal microscopy listserv, go to:
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*****

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Hello Jacques,
>
>How much power you get at the sample will depend
>to some degree on the light path and light
>delivery system of your specific microscope, in
>addition to the light source itself.
>
>The only LED system we have is the Insight SSI
>(solid state illuminator) on our DeltaVision
>system. It has LEDs rated from 22 mW (505-515
>nm) to 89 mW (563-588 nm). I am assuming these
>are nominal LED output power levels, although
>the brochure does not specify. For our lines, we
>measured 20 to 50 mW at the sample, or about
>35-50% of the rated power for any given line,
>which would be really good delivery efficiency
>(our laser scanning confocal is about 10%). This
>is several times the power levels we were
>getting with a conventional mercury bulb and
>excitation filters on a comparable (but not
>identical) system (about 6 to 20 mW depending on
>excitation filter used, if I remember
>correctly). For most wavelengths, this is as
>bright or brighter than our Exfo or Xenon
>illuminators, which are typically brighter (at
>the sample) than our older Hg bulb illuminators.
>However, these illumination systems are also
>installed on newer microscopes with possibly
>improved light delivery. We haven't had the
>system for long enough to encounter any
>potential issues, but we like the brightness and
>the speed of wavelength switching. Also, we do
>not have to worry about burned out excitation
>filters or UV leaking, and we do not have to
>worry about replacing bulbs, heat, or keeping
>track of when  lamps are turned on and off.
>Also, no worries about anyone forgetting to turn
>off the lamp on Friday night. I can't think of
>any downside at this time. My second choice in
>terms of convenience would be the Exfo or
>similar systems (long life and easy to replace
>bulbs, combined with good brightness).
>
>For reference, the InsightSSI specs can be found here:
>
>http://www.api.com/downloads/pdfs/lifescience/InsightSSI.pdf
>
>
>Julio Vazquez
>Fred Hutchinson Cancer Research Center
>Seattle, WA
>
>http://www.fhcrc.org
>
>==



Dear Julio,

Thank you so much for putting some actual
measurements into this important discussion.

Could I ask you for a few more details?

I assume that you did these tests with the same
objective, or at least with an objective with the
same NA and mag (and hence the same size and
location of the BFP. Light that hits the metal
will not pass through the objective). Assuming
that you measured light coming out of the
objective with some sort of photometer, the other
big variable is how you set the size of the field
diaphragm (referred to the image plane: For
instance, you might set it to be the same as the
field of view of a particular objective, or maybe
to some known and repeatable fraction thereof.)

Also, with respect to your comment on confocal
performance, I am assuming that the 10%
efficiency refers to the fraction of the light
leaving the laser that arrives at the focus
plane. Ten percent seems low and seems to suggest
to me that either the optics used to launch the
laser light into the fiber are not well aligned
or that the optics are set up to over-fill the
BFP of the particular objective used for the
test. Otherwise it is hard to see where so much
power would be lost: objective transmissions are
now usually quoted in the 80-90% range, and a
properly chosen dichroic should be about the same
while beam-steering mirrors and galvo mirrors
should be about 99%.

In any case, thank you for the numbers.

Regards,

Jim Pawley

***************************************************************************
Prof. James B. Pawley,                
Ph.  608-238-3953              
21. N. Prospect Ave. Madison, WI 53726 USA
[hidden email]
3D Microscopy of Living Cells Course, June 9-21, 2012, UBC, Vancouver Canada
Info: http://www.3dcourse.ubc.ca/ 
Application deadline 3/16/2012
               "If it ain't diffraction, it must be statistics." Anon. 11/16/12



>
>
>On Feb 22, 2012, at 2:07 AM, jacques wrote:
>
>>  *****
>>  To join, leave or search the confocal microscopy listserv, go to:
>>  http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>  *****
>>
>>  Hi all,
>>
>>  two years later, and a quite similar question.
>>
>>  I use a Zeiss epifluorescence microscope, and I'm thinking to replace my HBO
>>  housing by a LED multicolor source.
>>  Of course, Zeiss sells the colibri setup, but the choice on the market is
>>  growing day after day, and for example, Thorlabs also sells something with 4
>>  colors and a control unit, with the Zeiss mounting system.
>>  I'm sure many other manufacturers do the same.
>>
>>  Do anybody compared the HBO to a LED system for classical epifluorescence ?
>>  What should be the LED power to have something similar to a 100W at common
>>  wavelenghts (dapi, gfp, rhodamine, etc) ? Even after checking this page :
>>  http://www.olympusfluoview.com/theory/noncoherentsources.html
>>  It is still difficult to know which valeus and units should be considered...
>>
>>  Thanks for the help
>>
>>  J.
>>
>>
>>  -----
>>
>>  Jacques FATTACCIOLI
>>
>>  Département de Chimie
>>  Ecole Normale Supérieure
>>  24 rue Lhomond 75231 Paris Cedex 05
>>  Email : [hidden email]
>>  Web : http://jacquesfattaccioli.wordpress.com
>>  --
>>  View this message in context:
>>http://confocal-microscopy-list.588098.n2.nabble.com/LEDs-tp4283200p7307846.html
>>  Sent from the Confocal Microscopy List mailing list archive at Nabble.com.


--
***************************************************************************
Prof. James B. Pawley,                
Ph.  608-238-3953              
21. N. Prospect Ave. Madison, WI 53726 USA
[hidden email]
3D Microscopy of Living Cells Course, June 9-21, 2012, UBC, Vancouver Canada
Info: http://www.3dcourse.ubc.ca/ 
Application deadline 3/16/2012
               "If it ain't diffraction, it must be statistics." Anon. 11/16/12
Justin Ross Justin Ross
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Re: LEDs

In reply to this post by Julio Vazquez
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi,
We previously had the Xe lamps and changed to the SSI illumination on
our Deltavision Core. The exposure times were reduced by a factor of 5
to 10. The SSI is actually made by Lumencor (no commercial interest).
From my experience it works really well without the downsides of a lamp
of heat generation or having to align and change bulbs.
If you do decide to use LEDs, I strongly suggest still using bandpass
excitation filter (as suggested previously) because with some LEDs there
upto 5% light emission at wavelengths upto 50 nm higher than the peak
which will generally add a lot of background to your images. If you have
a spectrometer available I suggest scanning the emission to get the real
emission spectrum of the LED.


Regards,
Justin.

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of Julio Vazquez
Sent: Friday, 24 February 2012 4:14 AM
To: [hidden email]
Subject: Re: LEDs

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hello all,

Applied Precision just told me that the InsightSSI illuminator is
actually not LED based, so please ignore my previous posting, as far as
LEDs are concerned.

Julio.

==


On Feb 23, 2012, at 2:06 AM, jacques wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Thanks for your help and comments !
>
> Jacques
Boswell, Carl A - (cboswell) Boswell, Carl A - (cboswell)
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Re: LEDs

"If you have a spectrometer available I suggest scanning the emission to get the real emission spectrum of the LED."

It would be of more general use if the manufacturer supplied a standardized, trustworthy spectral output of these devices.  Then everyone could determine for themselves what would work best for their systems and samples without the need for additional instrumentation.
C


Carl A. Boswell
520-742-6131

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Justin Ross
Sent: Thursday, February 23, 2012 6:14 PM
To: [hidden email]
Subject: Re: LEDs

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi,
We previously had the Xe lamps and changed to the SSI illumination on our Deltavision Core. The exposure times were reduced by a factor of 5 to 10. The SSI is actually made by Lumencor (no commercial interest).
From my experience it works really well without the downsides of a lamp of heat generation or having to align and change bulbs.
If you do decide to use LEDs, I strongly suggest still using bandpass excitation filter (as suggested previously) because with some LEDs there upto 5% light emission at wavelengths upto 50 nm higher than the peak which will generally add a lot of background to your images. If you have a spectrometer available I suggest scanning the emission to get the real emission spectrum of the LED.


Regards,
Justin.

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of Julio Vazquez
Sent: Friday, 24 February 2012 4:14 AM
To: [hidden email]
Subject: Re: LEDs

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hello all,

Applied Precision just told me that the InsightSSI illuminator is actually not LED based, so please ignore my previous posting, as far as LEDs are concerned.

Julio.

==


On Feb 23, 2012, at 2:06 AM, jacques wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Thanks for your help and comments !
>
> Jacques

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