http://confocal-microscopy-list.275.s1.nabble.com/Safe-to-run-multiphoton-laser-at-100-power-for-overnight-continuous-imaging-tp7588726p7588741.html
In one of our imaging conditions with 10x dey objectives on live mouse we observed vapour condensation.. Thus making the image hazy ..
> On 26-Sep-2018, at 10:39 PM, Moulding, Dale <
[hidden email]> wrote:
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> Dear list,
>
> I've run the expt, decided to use a 10x0.3 as it's a lot cheaper than the dippers. We found we could get sufficient energy to the sample with a total of 3h30m continuous scanning. 2h at 700nm (1500mW), 1h30m at 800nm (3000mW).
> Good news, the objective didn't even get warm, no hint of damage.
> I destroyed my last power meter trying to measure the 2P laser (slightly exceeded it's limit!), but from memory with the Zeiss engineer meter only about 20% makes it to the sample. So 600mW perhaps. The objective has 90% transmission at 800nm.
>
> Cheers
>
> Dale
>
>
>
>
>
> Dale Moulding PhD FRMS
> ICH Light Microscopy Facility
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> -------- Original message --------
> From: "Armstrong, Brian" <
[hidden email]>
> Date: 26/09/2018 16:26 (GMT+00:00)
> To:
[hidden email]
> Subject: Re: Safe to run multiphoton laser at 100% power for overnight continuous imaging?
>
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> Dale, thank you for the answers. Can you please update this post when you complete the experiment? Especially if you damage the lens.
> Thanks in advance,
>
> Brian Armstrong PhD
> Associate Research Professor
> Developmental and Stem Cell Biology
> Diabetes and Metabolic Diseases
> Director, Light Microscopy Core
> Beckman Research Institute, City of Hope
>
>
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:
[hidden email]] On Behalf Of Moulding, Dale
> Sent: Wednesday, September 26, 2018 2:00 AM
> To:
[hidden email]
> Subject: Re: Safe to run multiphoton laser at 100% power for overnight continuous imaging?
>
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> Dear all,
>
> Thanks for the replies. It will be at 800nM, so full power output, and will be continuously firing though the objective for at least 4 hours as it is not for imaging but for catalysis.
> Zeiss aren't able to guarantee their objectives for this (drilling to the centre of the earth!), so if I do get a cracked lens / glue then I'm not sure our service contract will cover it.
>
> @Lorenzo, for your 16 hour experiment were you sending the laser through the objective non-stop for the entire time, or just imaging periodically?
> @Craig, agreed, it’s a lot of power, but not for imaging, for catalysis.
> @Brian, I love the 10x0.5 dipper for 2P work. Massive improvement over the 10x0.3.
> @Claire, I'm not sure Zeiss will cover the damage this time, and haven't really had a definitive answer, unless I've misunderstood them.
>
> Cheers
>
> Dale
>
> -----Original Message-----
> From: Confocal Microscopy List <
[hidden email]> On Behalf Of Craig Brideau
> Sent: 25 September 2018 17:13
> To:
[hidden email]
> Subject: Re: Safe to run multiphoton laser at 100% power for overnight continuous imaging?
>
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> You do raise a valid point Brian, the power of most Ti:Saphs varies fairly dramatically by wavelength. The transmission through the optical system will also be different, and what the system considers '100%' may not actually be that much power under certain situations. In most cases, however, a Ti:Saph can produce much more power than is necessary for imaging.
> Try measuring the power out of the objective when set to 100% to see what you are actually getting; maybe it isn't that much compared to the output of the laser once all the system losses are considered.
>
> Craig
>
>> On Tue, Sep 25, 2018 at 9:41 AM Armstrong, Brian <
[hidden email]> wrote:
>>
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http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy>> Post images on
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>>
>> Hi Dale, it depends in part on the wavelength. As I imagine you
>> already know the output power at say 1050nm is far lower than say
>> 800nm. I do not think the LASER "cares" what your output power is. As
>> for the optics I cannot say. I think that they fall off in
>> transmittance above a given wavelength in the IR range but I do not
>> know if this is because of a coating or properties of the lens??? I
>> have heard stories of people burning objectives. Pockels cells will also burn out after around 10 years or so.
>>
>> P.S. How do you like the 10x/0.5 W for 2P work?
>>
>> Brian Armstrong PhD
>> Associate Research Professor
>> Developmental and Stem Cell Biology
>> Diabetes and Metabolic Diseases
>> Director, Light Microscopy Core
>> Beckman Research Institute, City of Hope
>>
>>
>>
>> -----Original Message-----
>> From: Confocal Microscopy List
>> [mailto:
[hidden email]]
>> On Behalf Of Moulding, Dale
>> Sent: Tuesday, September 25, 2018 5:56 AM
>> To:
[hidden email]
>> Subject: Safe to run multiphoton laser at 100% power for overnight
>> continuous imaging?
>>
>> [Attention: This email came from an external source. Do not open
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>>
>>
>>
>>
>> *****
>> 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.
>> *****
>>
>> Dear list,
>>
>> I've been asked to run our 2 photon laser (Mai Tai eHP DeepSee) at
>> full power (3W) non-stop over night to scan a large sample.
>> When doing ablations we run at 100% and I hadn't even given it a
>> second thought, as the ablations are over in a flash.
>> However, continuously (4 to 16 hours) running 100% 800nM at 3W I'm
>> concerned we may damage the optics of the system. It will be on a
>> Zeiss Examiner, going through Zeiss's 10x0.5 or 20x1.0 water dippers.
>> Has anyone ever imaged / ablated with a 2P laser continuously at full
>> power?
>> Should I be concerned that the objectives may be damaged?
>>
>> Cheers
>>
>> Dale
>>
>>
>> Dale Moulding PhD FRMS
>> ICH Light Microscopy Facility
>> Room W2.06, BDRC office
>> UCL Great Ormond Street Institute of Child Health
>> 30 Guilford St
>> London WC1N 1EH
>> Mob: 07787 699609
>> Tel: 020 7905 2753
>>
>>
http://www.ucl.ac.uk/ich/core-scientific-facilities-centres/confocal-m>> icroscopy
>>
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