LED light sources for live cell imaging

[SUBSCRIBE CONFOCALMICROSCOPY Anonymous-3]

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Dear All,

While defining specs for a live cell microscope we are looking at the LED light sources available in the market. There are Lumencor, CoolLED, X-Cite and Colibri.7. Is there any comparative table available in the list?

Best regards,
Core Imaging Facility Staff
NTU

[Sylvie Le Guyader]

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Xcite is a metal Halide lamp, not Led. To image with that you need a slow shutter. If you only plan on using it to peep in the eyepiece, it is just fine. However I would include in my thinking an eventual future upgrade of your confocal to getting a side camera for fast widefield. With LEDs you already have a fast illumination /shutter system in place. If you only have 1 confocal, you might want to keep it dedicated but if you have several, it is a good way to get a 'cheap' widefield.

We only have some experience with Lumencore Spectrax and are happy with it.

Good luck

Med vänlig hälsning / Best regards

Sylvie

@@@@@@@@@@@@@@@@@@@@@@@@

Sylvie Le Guyader, PhD
Live Cell Imaging Facility Manager
Karolinska Institutet- Bionut Dpt
Hälsovägen 7,
Novum, G lift, floor 6
14157<tel:14157> Huddinge
Sweden
mobile: [phone]<tel:+46%20(0)%2073%20733%205008>
office: [phone]<tel:+46%20(0)%2008-524%20811%2072>
LCI website

---- SUBSCRIBE CONFOCALMICROSCOPY Anonymous wrote ----

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Dear All,

While defining specs for a live cell microscope we are looking at the LED light sources available in the market. There are Lumencor, CoolLED, X-Cite and Colibri.7. Is there any comparative table available in the list?

Best regards,
Core Imaging Facility Staff
NTU

[Simon Kinder]

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I think they perhaps mean this

http://www.excelitas.com/Pages/Product/X-Cite-XLED1.aspx

Which is LED but also part of the X-Cite range

Simon


Simon Kinder
Product Manager

OLYMPUS AUSTRALIA
Ground Floor
82 Waterloo Road, North Ryde, New South Wales, 2113


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-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Sylvie Le Guyader
Sent: Monday, 24 July 2017 5:18 PM
To: [hidden email]
Subject: Re: LED light sources for live cell imaging

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Xcite is a metal Halide lamp, not Led. To image with that you need a slow shutter. If you only plan on using it to peep in the eyepiece, it is just fine. However I would include in my thinking an eventual future upgrade of your confocal to getting a side camera for fast widefield. With LEDs you already have a fast illumination /shutter system in place. If you only have 1 confocal, you might want to keep it dedicated but if you have several, it is a good way to get a 'cheap' widefield.

We only have some experience with Lumencore Spectrax and are happy with it.

Good luck

Med vänlig hälsning / Best regards

Sylvie

@@@@@@@@@@@@@@@@@@@@@@@@

Sylvie Le Guyader, PhD
Live Cell Imaging Facility Manager
Karolinska Institutet- Bionut Dpt
Hälsovägen 7,
Novum, G lift, floor 6
14157<tel:14157> Huddinge
Sweden
mobile: [phone]<tel:+46%20(0)%2073%20733%205008>
office: [phone]<tel:+46%20(0)%2008-524%20811%2072>
LCI website

---- SUBSCRIBE CONFOCALMICROSCOPY Anonymous wrote ----

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Dear All,

While defining specs for a live cell microscope we are looking at the LED light sources available in the market. There are Lumencor, CoolLED, X-Cite and Colibri.7. Is there any comparative table available in the list?

Best regards,
Core Imaging Facility Staff
NTU
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[Claire Brown]

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We have had good success with the Lumencor Sola XM, XCite 120 LED and CoolLED pE300white. We have a Colibri system but it is on a scope we don't do live cell imaging on. I don't know of any comparison list but here are a  few general comments.

1) We have moved towards direct coupling to the microscope when possible. We have had issues with the liquid light guides. They often get bubbles in them that cause black shadows in the image. Also the coupling sometimes makes it hard to get a good uniform field of view. It is just one more part to worry about and possibly have to have the expense to replace.

2) In my experience all of these light sources are way too bright for live cell imaging. We are using them at less than 1% power even for the ones that are "dim" in the blue region where EGFP is excited. We use ND filters to reduce the light even further when we can. We have a paper that will come out soon showing that putting the light intensity down and the exposure time up - even to 1 s if needed - really improves cell health. So short bursts of bright light (we tested down to 17 ms exposures) can cause photo damage when the same light dose of low intensity light for a long exposure time will not cause any visible damage. I have been pushing the manufacturers to come out with a much dimmer (lower cost, lower power consumption) option for live cells but the industry trend is to get brighter and brighter to open these light sources to other applications like spinning disk confocal for example. We even tried the CoolLED pe100 transmitted light source for live cell fluorescence imaging. It works well but it can't be used for blue dye excitation so would not be ideal on a system where you might want to look at DAPI on fixed cells on occasion as well.

3) I would not recommend the metal-halide systems unless you need to go down into the UV for excitation. The bulbs are expensive, have to be changed taking up staff time and they contain mercury. So the LEDs are more sustainable.

4) My current favorite is the pE300 white. The price is good, it is very compact and it can be direct coupled to the microscope.

I have no commercial interest in any with any of these companies these are just my general impressions from working with the different light sources.

Good luck!

Claire

[SUBSCRIBE CONFOCALMICROSCOPY Anonymous-3]

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Dear Claire,

Thanks much for a detailed info on the LED light sources for microscopy.
LEDs have below 400 nm lines. Are they not efficient in exciting UV labels?
I do not understand what you meant by metal-halide for UV excitation.

Is it Colibri.2 or Colibri.7? I heard that Colibri.7 is much better than
the previous ones.

Best regards,


On 26 July 2017 at 22:06, Claire Brown <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> We have had good success with the Lumencor Sola XM, XCite 120 LED and
> CoolLED pE300white. We have a Colibri system but it is on a scope we don't
> do live cell imaging on. I don't know of any comparison list but here are
> a  few general comments.
>
> 1) We have moved towards direct coupling to the microscope when possible.
> We have had issues with the liquid light guides. They often get bubbles in
> them that cause black shadows in the image. Also the coupling sometimes
> makes it hard to get a good uniform field of view. It is just one more part
> to worry about and possibly have to have the expense to replace.
>
> 2) In my experience all of these light sources are way too bright for live
> cell imaging. We are using them at less than 1% power even for the ones
> that are "dim" in the blue region where EGFP is excited. We use ND filters
> to reduce the light even further when we can. We have a paper that will
> come out soon showing that putting the light intensity down and the
> exposure time up - even to 1 s if needed - really improves cell health. So
> short bursts of bright light (we tested down to 17 ms exposures) can cause
> photo damage when the same light dose of low intensity light for a long
> exposure time will not cause any visible damage. I have been pushing the
> manufacturers to come out with a much dimmer (lower cost, lower power
> consumption) option for live cells but the industry trend is to get
> brighter and brighter to open these light sources to other applications
> like spinning disk confocal for example. We even tried the CoolLED pe100
> transmitted light source for live cell fluorescence imaging. It works well
> but it can't be used for blue dye excitation so would not be ideal on a
> system where you might want to look at DAPI on fixed cells on occasion as
> well.
>
> 3) I would not recommend the metal-halide systems unless you need to go
> down into the UV for excitation. The bulbs are expensive, have to be
> changed taking up staff time and they contain mercury. So the LEDs are more
> sustainable.
>
> 4) My current favorite is the pE300 white. The price is good, it is very
> compact and it can be direct coupled to the microscope.
>
> I have no commercial interest in any with any of these companies these are
> just my general impressions from working with the different light sources.
>
> Good luck!
>
> Claire
>

[Claire Brown]

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Metal halides go down to about 300 nm. The LEDs below 370 or so are pretty new so we have not tested them.

We have used the earlier Colibri models. I'm not sure if ours is the 2. We have not tried the 7 yet.

Claire

[Steffen Dietzel]

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Am 26.07.2017 um 16:06 schrieb Claire Brown
> 3) I would not recommend the metal-halide systems unless you need to go down into the UV for excitation. The bulbs are expensive, have to be changed taking up staff time and they contain mercury. So the LEDs are more sustainable.
>

Plus, Mercury produces a pretty strong UV-Peak which cannot be filtered
out 100% and which tends to kill cells, at least at higher magnification
(60-100). Before LEDs were available, we therefore used a normal halogen
lamp for fluorescence excitation. Needed long exposure times, but the
cells were happy. With a T-house one could have had both on the stand at
the same time, but we went for the cheaper, screwdriver solution.

Not even starting about the hassle of having to leave a Mercury or
Mercury-halide system at least half an hour on and at least half an hour
off, when switched. Or about environmental concerns with regard to Mercury.

Steffen

--
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Ludwig-Maximilians-Universität München
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