objective lens laser damage

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We damaged an objective lens, a Nikon CFI Plan Apo Lambda 100X Oil 1.45 NA
lens on our homebuilt TIRF/STORM microscope. Has anybody seen this sort of
thing before? I'm interested to hear, if so. We will repair or replace it,
and if replacing I wonder what to get. More details are below.

Here is a link to a pic of part of the back focal plane. It has some dark
spots that look 'burned' and some stuff that looks like 'bubbles'.

http://i63.tinypic.com/mmemvl.jpg


Repairing this lens will cost >50% of a new lens so I want to decide whether
to repair or to just get something else. I'd prefer something with a higher
damage threshold but I was unsuccessful in getting data on lens damage
thresholds.

If replacing, what to get? I noticed three lenses online and I wonder what
is best for STORM and whether all have a similar damage threshold or not.

1. CFI Plan Apo Lambda 100X Oil 1.45 NA (current lens)
2. CFI Plan Apo Lambda DM 100X Oil 1.45 NA
3. CFI Apo SR TIRF 100X Oil 1.49 NA


I think the Lambda lenses are supposed to be lower background, though I've
never tested whether that is important or not. They are also Plan lenses,
but I'm not sure how important that is for the <40 micron regions we
typically image. The SR lens has a correction collar which might be useful
(?) when imaging a bit off the glass. The online information says the SR
lens has a PSF optimized for STORM which sounds good but I'm not sure what
it actually means. I don't have any damage threshold information about these
lenses. Oh, we are pretty sure the damage was caused when we sent ~120 mW at
405 nm was sent to the lens in EPI and later in TIRF. We plan to
substantially attenuate the laser in the future, which didn't need to be
that big anyway, but I am not sure how much to attenuate by.


Joshua C. Vaughan
Assistant Professor
Department of Chemistry
Box 351700
University of Washington
Seattle, WA 98195

[Seamus Holden-2]

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Nikon have started selling a "high power" version of their 100x 1.49 TIRF objective designed for use with their latest NSTORM system, but presumably purchasable separately. From conversation, the glass is apparently optimized for particularly high transmission, I guess in order to reduce energy transfer to the lens at the BFP.
Info is here:
https://www.nikoninstruments.com/en_GB/Products/Super-resolution/N-STORM-4.0-Super-Resolution
These HP objectives are different to the SR lenses - which I have one of - which have an optimal PSF but no high power optimization - they are designed for SIM basically.

I am interested what laser power and wavelength this damage occurred at. In the past I have routinely focussed 150 mW of either 642 nm or 561 nm illumination into the BFP of my objective for STORM/ PALM and did not run into problems. Although of course it could be possible I did not notice? How did the BFP damage present as in the image plane? Increased PSF aberration, lower transmission?

Best wishes

Dr Seamus Holden
University Research Fellow

Centre for Bacterial Cell Biology
Baddiley-Clark Building
Newcastle University
Richardson Road
Newcastle upon Tyne
NE2 4AX, United Kingdom

Phone: [phone]

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Joshua Vaughan
Sent: 27 May 2016 08:59
To: [hidden email]
Subject: objective lens laser damage

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We damaged an objective lens, a Nikon CFI Plan Apo Lambda 100X Oil 1.45 NA lens on our homebuilt TIRF/STORM microscope. Has anybody seen this sort of thing before? I'm interested to hear, if so. We will repair or replace it, and if replacing I wonder what to get. More details are below.

Here is a link to a pic of part of the back focal plane. It has some dark spots that look 'burned' and some stuff that looks like 'bubbles'.

http://i63.tinypic.com/mmemvl.jpg


Repairing this lens will cost >50% of a new lens so I want to decide whether to repair or to just get something else. I'd prefer something with a higher damage threshold but I was unsuccessful in getting data on lens damage thresholds.

If replacing, what to get? I noticed three lenses online and I wonder what is best for STORM and whether all have a similar damage threshold or not.

1. CFI Plan Apo Lambda 100X Oil 1.45 NA (current lens) 2. CFI Plan Apo Lambda DM 100X Oil 1.45 NA 3. CFI Apo SR TIRF 100X Oil 1.49 NA


I think the Lambda lenses are supposed to be lower background, though I've never tested whether that is important or not. They are also Plan lenses, but I'm not sure how important that is for the <40 micron regions we typically image. The SR lens has a correction collar which might be useful
(?) when imaging a bit off the glass. The online information says the SR lens has a PSF optimized for STORM which sounds good but I'm not sure what it actually means. I don't have any damage threshold information about these lenses. Oh, we are pretty sure the damage was caused when we sent ~120 mW at
405 nm was sent to the lens in EPI and later in TIRF. We plan to substantially attenuate the laser in the future, which didn't need to be that big anyway, but I am not sure how much to attenuate by.


Joshua C. Vaughan
Assistant Professor
Department of Chemistry
Box 351700
University of Washington
Seattle, WA 98195

[Martin Wessendorf-2]

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Dear Dr. Vaughan--

Could it be dirt and oil on the lens, rather than being burned and/or
unglued?

Martin Wessendorf



On 5/27/2016 2:58 AM, Joshua Vaughan 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 damaged an objective lens, a Nikon CFI Plan Apo Lambda 100X Oil 1.45 NA
> lens on our homebuilt TIRF/STORM microscope. Has anybody seen this sort of
> thing before? I'm interested to hear, if so. We will repair or replace it,
> and if replacing I wonder what to get. More details are below.
>
> Here is a link to a pic of part of the back focal plane. It has some dark
> spots that look 'burned' and some stuff that looks like 'bubbles'.
>
> http://i63.tinypic.com/mmemvl.jpg
>
>
> Repairing this lens will cost >50% of a new lens so I want to decide whether
> to repair or to just get something else. I'd prefer something with a higher
> damage threshold but I was unsuccessful in getting data on lens damage
> thresholds.
>
> If replacing, what to get? I noticed three lenses online and I wonder what
> is best for STORM and whether all have a similar damage threshold or not.
>
> 1. CFI Plan Apo Lambda 100X Oil 1.45 NA (current lens)
> 2. CFI Plan Apo Lambda DM 100X Oil 1.45 NA
> 3. CFI Apo SR TIRF 100X Oil 1.49 NA
>
>
> I think the Lambda lenses are supposed to be lower background, though I've
> never tested whether that is important or not. They are also Plan lenses,
> but I'm not sure how important that is for the <40 micron regions we
> typically image. The SR lens has a correction collar which might be useful
> (?) when imaging a bit off the glass. The online information says the SR
> lens has a PSF optimized for STORM which sounds good but I'm not sure what
> it actually means. I don't have any damage threshold information about these
> lenses. Oh, we are pretty sure the damage was caused when we sent ~120 mW at
> 405 nm was sent to the lens in EPI and later in TIRF. We plan to
> substantially attenuate the laser in the future, which didn't need to be
> that big anyway, but I am not sure how much to attenuate by.
>
>
> Joshua C. Vaughan
> Assistant Professor
> Department of Chemistry
> Box 351700
> University of Washington
> Seattle, WA 98195

--
Martin Wessendorf, Ph.D.                   office: [phone]
Assoc Prof, Dept Neuroscience                 lab: [phone]
University of Minnesota             Preferred FAX: [phone]
6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
Minneapolis, MN  55455                    e-mail: [hidden email]

[Craig Brideau]

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I had a glance at your picture. The black spots look like laser burns, but
there is also a clear bubble or edge that looks suspiciously like oil also
visible in the picture. It's possible you had some oil contamination and
then it burned onto the glass. Most optics will handle a surprising amount
of laser power as long as they are extremely clean. The instant there is
any dust/grease the laser will burn the surface as the contamination
increases point absorption at the interface.

Craig

On Fri, May 27, 2016 at 7:28 AM, Martin Wessendorf <[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.
> *****
>
> Dear Dr. Vaughan--
>
> Could it be dirt and oil on the lens, rather than being burned and/or
> unglued?
>
> Martin Wessendorf
>
>
>
>
> On 5/27/2016 2:58 AM, Joshua Vaughan 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 damaged an objective lens, a Nikon CFI Plan Apo Lambda 100X Oil 1.45 NA
>> lens on our homebuilt TIRF/STORM microscope. Has anybody seen this sort of
>> thing before? I'm interested to hear, if so. We will repair or replace it,
>> and if replacing I wonder what to get. More details are below.
>>
>> Here is a link to a pic of part of the back focal plane. It has some dark
>> spots that look 'burned' and some stuff that looks like 'bubbles'.
>>
>> http://i63.tinypic.com/mmemvl.jpg
>>
>>
>> Repairing this lens will cost >50% of a new lens so I want to decide
>> whether
>> to repair or to just get something else. I'd prefer something with a
>> higher
>> damage threshold but I was unsuccessful in getting data on lens damage
>> thresholds.
>>
>> If replacing, what to get? I noticed three lenses online and I wonder what
>> is best for STORM and whether all have a similar damage threshold or not.
>>
>> 1. CFI Plan Apo Lambda 100X Oil 1.45 NA (current lens)
>> 2. CFI Plan Apo Lambda DM 100X Oil 1.45 NA
>> 3. CFI Apo SR TIRF 100X Oil 1.49 NA
>>
>>
>> I think the Lambda lenses are supposed to be lower background, though I've
>> never tested whether that is important or not. They are also Plan lenses,
>> but I'm not sure how important that is for the <40 micron regions we
>> typically image. The SR lens has a correction collar which might be useful
>> (?) when imaging a bit off the glass. The online information says the SR
>> lens has a PSF optimized for STORM which sounds good but I'm not sure what
>> it actually means. I don't have any damage threshold information about
>> these
>> lenses. Oh, we are pretty sure the damage was caused when we sent ~120 mW
>> at
>> 405 nm was sent to the lens in EPI and later in TIRF. We plan to
>> substantially attenuate the laser in the future, which didn't need to be
>> that big anyway, but I am not sure how much to attenuate by.
>>
>>
>> Joshua C. Vaughan
>> Assistant Professor
>> Department of Chemistry
>> Box 351700
>> University of Washington
>> Seattle, WA 98195
>>
>
> --
> Martin Wessendorf, Ph.D.                   office: [phone]
> Assoc Prof, Dept Neuroscience                 lab: [phone]
> University of Minnesota             Preferred FAX: [phone]
> 6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
> Minneapolis, MN  55455                    e-mail: [hidden email]
>

[Nuno Moreno]

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*****

Dear Craig

I sent already a message directly to Joshua with a similar statement. The problem is however when the burning in not at the first lens but somewhere inside. In this case this means that the objective was not assembled in a proper clean environment and has dust or other particles inside. Actually here at the institute we were surprised on how often this happens from 2 different manufacturers. Maybe we were just unlucky. If that is the case I would ask for a replacement free of charge. I must confess however that in Joshua case it does not look like but the picture is not clear enough. I would advice you to bring your objective to a stereoscope, start to focus on the first back optical element and check whether this is superficial or not.

Kind regards and good luck
Nuno Moreno

> On 27 May 2016, at 15:07, Craig Brideau <[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.
> *****
>
> I had a glance at your picture. The black spots look like laser burns, but
> there is also a clear bubble or edge that looks suspiciously like oil also
> visible in the picture. It's possible you had some oil contamination and
> then it burned onto the glass. Most optics will handle a surprising amount
> of laser power as long as they are extremely clean. The instant there is
> any dust/grease the laser will burn the surface as the contamination
> increases point absorption at the interface.
>
> Craig
>
> On Fri, May 27, 2016 at 7:28 AM, Martin Wessendorf <[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.
>> *****
>>
>> Dear Dr. Vaughan--
>>
>> Could it be dirt and oil on the lens, rather than being burned and/or
>> unglued?
>>
>> Martin Wessendorf
>>
>>
>>
>>
>> On 5/27/2016 2:58 AM, Joshua Vaughan 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 damaged an objective lens, a Nikon CFI Plan Apo Lambda 100X Oil 1.45 NA
>>> lens on our homebuilt TIRF/STORM microscope. Has anybody seen this sort of
>>> thing before? I'm interested to hear, if so. We will repair or replace it,
>>> and if replacing I wonder what to get. More details are below.
>>>
>>> Here is a link to a pic of part of the back focal plane. It has some dark
>>> spots that look 'burned' and some stuff that looks like 'bubbles'.
>>>
>>> http://i63.tinypic.com/mmemvl.jpg
>>>
>>>
>>> Repairing this lens will cost >50% of a new lens so I want to decide
>>> whether
>>> to repair or to just get something else. I'd prefer something with a
>>> higher
>>> damage threshold but I was unsuccessful in getting data on lens damage
>>> thresholds.
>>>
>>> If replacing, what to get? I noticed three lenses online and I wonder what
>>> is best for STORM and whether all have a similar damage threshold or not.
>>>
>>> 1. CFI Plan Apo Lambda 100X Oil 1.45 NA (current lens)
>>> 2. CFI Plan Apo Lambda DM 100X Oil 1.45 NA
>>> 3. CFI Apo SR TIRF 100X Oil 1.49 NA
>>>
>>>
>>> I think the Lambda lenses are supposed to be lower background, though I've
>>> never tested whether that is important or not. They are also Plan lenses,
>>> but I'm not sure how important that is for the <40 micron regions we
>>> typically image. The SR lens has a correction collar which might be useful
>>> (?) when imaging a bit off the glass. The online information says the SR
>>> lens has a PSF optimized for STORM which sounds good but I'm not sure what
>>> it actually means. I don't have any damage threshold information about
>>> these
>>> lenses. Oh, we are pretty sure the damage was caused when we sent ~120 mW
>>> at
>>> 405 nm was sent to the lens in EPI and later in TIRF. We plan to
>>> substantially attenuate the laser in the future, which didn't need to be
>>> that big anyway, but I am not sure how much to attenuate by.
>>>
>>>
>>> Joshua C. Vaughan
>>> Assistant Professor
>>> Department of Chemistry
>>> Box 351700
>>> University of Washington
>>> Seattle, WA 98195
>>>
>>
>> --
>> Martin Wessendorf, Ph.D.                   office: [phone]
>> Assoc Prof, Dept Neuroscience                 lab: [phone]
>> University of Minnesota             Preferred FAX: [phone]
>> 6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
>> Minneapolis, MN  55455                    e-mail: [hidden email]
>>

[Zdenek Svindrych-2]

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Hi Joshua,
does not look good at all! Too bad...


We've been focusing some 200 mW at 480nm and even more at 532 nm without any
problems, but the chance of damaging lens depends on wavelength (of course
405 nm will be much worse) and on specific focusing of the beam. In TIRF you
usually have diffraction-limited spot in the BFP, if you focus on a lens
element, or even worse on a glue in a doublet or triplet, then ...

Indeed, the bubbles look like glue between lenses that have partially
separated. Sort of really, really bad luck. This can happen with any
widefield illumination with gaussian (diffraction-limited) beam, including
SIM and TIRF, but I've actually never seen that before...

In a "high power handling" lens I would avoid any glued lens multiplets, but
who knows what the vendors do.

Best of luck to your remaining eye (and objective lenses), zdenek


--
Zdenek Svindrych, Ph.D.
W.M. Keck Center for Cellular Imaging (PLSB 003)
University of Virginia, Charlottesville, VA
http://www.kcci.virginia.edu/
tel: [phone]
Annual FRET Workshop: http://kcci.virginia.edu/workshop-2017



---------- Původní zpráva ----------
Od: Joshua Vaughan <[hidden email]>
Komu: [hidden email]
Datum: 27. 5. 2016 4:11:25
Předmět: objective lens laser damage

"*****
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Post images on http://www.imgur.com and include the link in your posting.
*****

We damaged an objective lens, a Nikon CFI Plan Apo Lambda 100X Oil 1.45 NA
lens on our homebuilt TIRF/STORM microscope. Has anybody seen this sort of
thing before? I'm interested to hear, if so. We will repair or replace it,
and if replacing I wonder what to get. More details are below.

Here is a link to a pic of part of the back focal plane. It has some dark
spots that look 'burned' and some stuff that looks like 'bubbles'.

http://i63.tinypic.com/mmemvl.jpg


Repairing this lens will cost >50% of a new lens so I want to decide whether
to repair or to just get something else. I'd prefer something with a higher
damage threshold but I was unsuccessful in getting data on lens damage
thresholds.

If replacing, what to get? I noticed three lenses online and I wonder what
is best for STORM and whether all have a similar damage threshold or not.

1. CFI Plan Apo Lambda 100X Oil 1.45 NA (current lens)
2. CFI Plan Apo Lambda DM 100X Oil 1.45 NA
3. CFI Apo SR TIRF 100X Oil 1.49 NA


I think the Lambda lenses are supposed to be lower background, though I've
never tested whether that is important or not. They are also Plan lenses,
but I'm not sure how important that is for the <40 micron regions we
typically image. The SR lens has a correction collar which might be useful
(?) when imaging a bit off the glass. The online information says the SR
lens has a PSF optimized for STORM which sounds good but I'm not sure what
it actually means. I don't have any damage threshold information about these
lenses. Oh, we are pretty sure the damage was caused when we sent ~120 mW at
405 nm was sent to the lens in EPI and later in TIRF. We plan to
substantially attenuate the laser in the future, which didn't need to be
that big anyway, but I am not sure how much to attenuate by.


Joshua C. Vaughan
Assistant Professor
Department of Chemistry
Box 351700
University of Washington
Seattle, WA 98195"

[Glen MacDonald-2]

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Possibly the damage is to the optical cement between the lens elements.  I found epoxy to be very absorbtive and could be damaged during 2-photon imaging.  

Regards,
Glen MacDonald
Digital Microscopy Center
Box 357923
University of Washington
Seattle, WA 98195-7923  USA
[phone]
[hidden email]

> On May 27, 2016, at 8:03 AM, Nuno Moreno <[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.
> *****
>
> Dear Craig
>
> I sent already a message directly to Joshua with a similar statement. The problem is however when the burning in not at the first lens but somewhere inside. In this case this means that the objective was not assembled in a proper clean environment and has dust or other particles inside. Actually here at the institute we were surprised on how often this happens from 2 different manufacturers. Maybe we were just unlucky. If that is the case I would ask for a replacement free of charge. I must confess however that in Joshua case it does not look like but the picture is not clear enough. I would advice you to bring your objective to a stereoscope, start to focus on the first back optical element and check whether this is superficial or not.
>
> Kind regards and good luck
> Nuno Moreno
>
>> On 27 May 2016, at 15:07, Craig Brideau <[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.
>> *****
>>
>> I had a glance at your picture. The black spots look like laser burns, but
>> there is also a clear bubble or edge that looks suspiciously like oil also
>> visible in the picture. It's possible you had some oil contamination and
>> then it burned onto the glass. Most optics will handle a surprising amount
>> of laser power as long as they are extremely clean. The instant there is
>> any dust/grease the laser will burn the surface as the contamination
>> increases point absorption at the interface.
>>
>> Craig
>>
>> On Fri, May 27, 2016 at 7:28 AM, Martin Wessendorf <[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.
>>> *****
>>>
>>> Dear Dr. Vaughan--
>>>
>>> Could it be dirt and oil on the lens, rather than being burned and/or
>>> unglued?
>>>
>>> Martin Wessendorf
>>>
>>>
>>>
>>>
>>> On 5/27/2016 2:58 AM, Joshua Vaughan 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 damaged an objective lens, a Nikon CFI Plan Apo Lambda 100X Oil 1.45 NA
>>>> lens on our homebuilt TIRF/STORM microscope. Has anybody seen this sort of
>>>> thing before? I'm interested to hear, if so. We will repair or replace it,
>>>> and if replacing I wonder what to get. More details are below.
>>>>
>>>> Here is a link to a pic of part of the back focal plane. It has some dark
>>>> spots that look 'burned' and some stuff that looks like 'bubbles'.
>>>>
>>>> http://i63.tinypic.com/mmemvl.jpg
>>>>
>>>>
>>>> Repairing this lens will cost >50% of a new lens so I want to decide
>>>> whether
>>>> to repair or to just get something else. I'd prefer something with a
>>>> higher
>>>> damage threshold but I was unsuccessful in getting data on lens damage
>>>> thresholds.
>>>>
>>>> If replacing, what to get? I noticed three lenses online and I wonder what
>>>> is best for STORM and whether all have a similar damage threshold or not.
>>>>
>>>> 1. CFI Plan Apo Lambda 100X Oil 1.45 NA (current lens)
>>>> 2. CFI Plan Apo Lambda DM 100X Oil 1.45 NA
>>>> 3. CFI Apo SR TIRF 100X Oil 1.49 NA
>>>>
>>>>
>>>> I think the Lambda lenses are supposed to be lower background, though I've
>>>> never tested whether that is important or not. They are also Plan lenses,
>>>> but I'm not sure how important that is for the <40 micron regions we
>>>> typically image. The SR lens has a correction collar which might be useful
>>>> (?) when imaging a bit off the glass. The online information says the SR
>>>> lens has a PSF optimized for STORM which sounds good but I'm not sure what
>>>> it actually means. I don't have any damage threshold information about
>>>> these
>>>> lenses. Oh, we are pretty sure the damage was caused when we sent ~120 mW
>>>> at
>>>> 405 nm was sent to the lens in EPI and later in TIRF. We plan to
>>>> substantially attenuate the laser in the future, which didn't need to be
>>>> that big anyway, but I am not sure how much to attenuate by.
>>>>
>>>>
>>>> Joshua C. Vaughan
>>>> Assistant Professor
>>>> Department of Chemistry
>>>> Box 351700
>>>> University of Washington
>>>> Seattle, WA 98195
>>>>
>>>
>>> --
>>> Martin Wessendorf, Ph.D.                   office: [phone]
>>> Assoc Prof, Dept Neuroscience                 lab: [phone]
>>> University of Minnesota             Preferred FAX: [phone]
>>> 6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
>>> Minneapolis, MN  55455                    e-mail: [hidden email]
>>>

*****
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Followup to some replies.

Seamus:
The various damage spots happened over a period of the last ~2 years. I
have relatively large lasers, with 405/488/561/647/750 at
~200/1000/2000/1500/500 mW but we generally run at much lower powers. This
is more laser power than most, I know, but I used something like this
previously on an Olympus UPlanSApo 1.4 NA oil lens that never had trouble
so I thought it would be ok. We noticed the damage first in the
illumination since in 'straight epi' with the beam going up we got lots of
scattering and uneven illumination so we got into the habit of aligning
(with very low powers!) slightly off straight epi. Over time there was more
damage, particularly near TIRF positions on either side of the lens. At
this point, the PSF is noticeably asymmetric and starting to hurt things
for 3D by astigmatism, though for 2D near focus things were ok-ish.

Martin, Nuno, Craig:
I can't say for sure whether the lens had gotten dirt or oil inside though
I believe not. My group is generally careful, but I suppose someone could
have gotten oil down the barrel of the Nikon lens (the Olympus UIS2
mushroom-cap type design is better for keeping out oil). The image I posted
has is focused roughly on the back focal plane so I believe the issue is
interior to the objective. The back hemisphere of the lens looked fine to
me when I examined reflections carefully near a bright light.

Zdenek:
Before sending the lens back, we did a couple tests in an initially
undamaged region that showed no damage in the back focal plane. Lots of 647
nm power (~1000 mW) did not lead to damage in the test, but lots of 405 nm
power (~100 mW) led to a new and prominent 'bubble' being formed around the
illuminated part of the BFP within 10-20 minutes. I figured if we were
going to replace or repair we might as well try to figure out a little more
information about the culprit so we can try to avoid this problem in the
future.

A little more info:
Folks at my old postdoc lab tell me that they also see damage in their 60x
version of the Lambda lens (I think it is CFI Plan Apo Lambda 60x Oil
1.4NA) although it is not as dramatic and that lens has been used for a
shorter period than mine (~3 years of use). I wonder whether there are more
damaged lenses out there that go unnoticed since it is a little hard to
tell in TIRF and since I suspect few people look at the back focal plane.
When I inspect, in addition to illuminating with the relatively low NA
condenser like in the posted photo, sometimes I use epi-illumination of
beads to fill the back aperture.



On Fri, May 27, 2016 at 8:03 AM, Nuno Moreno <[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.
> *****
>
> Dear Craig
>
> I sent already a message directly to Joshua with a similar statement. The
> problem is however when the burning in not at the first lens but somewhere
> inside. In this case this means that the objective was not assembled in a
> proper clean environment and has dust or other particles inside. Actually
> here at the institute we were surprised on how often this happens from 2
> different manufacturers. Maybe we were just unlucky. If that is the case I
> would ask for a replacement free of charge. I must confess however that in
> Joshua case it does not look like but the picture is not clear enough. I
> would advice you to bring your objective to a stereoscope, start to focus
> on the first back optical element and check whether this is superficial or
> not.
>
> Kind regards and good luck
> Nuno Moreno
>
> > On 27 May 2016, at 15:07, Craig Brideau <[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.
> > *****
> >
> > I had a glance at your picture. The black spots look like laser burns,
> but
> > there is also a clear bubble or edge that looks suspiciously like oil
> also
> > visible in the picture. It's possible you had some oil contamination and
> > then it burned onto the glass. Most optics will handle a surprising
> amount
> > of laser power as long as they are extremely clean. The instant there is
> > any dust/grease the laser will burn the surface as the contamination
> > increases point absorption at the interface.
> >
> > Craig
> >
> > On Fri, May 27, 2016 at 7:28 AM, Martin Wessendorf <[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.
> >> *****
> >>
> >> Dear Dr. Vaughan--
> >>
> >> Could it be dirt and oil on the lens, rather than being burned and/or
> >> unglued?
> >>
> >> Martin Wessendorf
> >>
> >>
> >>
> >>
> >> On 5/27/2016 2:58 AM, Joshua Vaughan 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 damaged an objective lens, a Nikon CFI Plan Apo Lambda 100X Oil
> 1.45 NA
> >>> lens on our homebuilt TIRF/STORM microscope. Has anybody seen this
> sort of
> >>> thing before? I'm interested to hear, if so. We will repair or replace
> it,
> >>> and if replacing I wonder what to get. More details are below.
> >>>
> >>> Here is a link to a pic of part of the back focal plane. It has some
> dark
> >>> spots that look 'burned' and some stuff that looks like 'bubbles'.
> >>>
> >>> http://i63.tinypic.com/mmemvl.jpg
> >>>
> >>>
> >>> Repairing this lens will cost >50% of a new lens so I want to decide
> >>> whether
> >>> to repair or to just get something else. I'd prefer something with a
> >>> higher
> >>> damage threshold but I was unsuccessful in getting data on lens damage
> >>> thresholds.
> >>>
> >>> If replacing, what to get? I noticed three lenses online and I wonder
> what
> >>> is best for STORM and whether all have a similar damage threshold or
> not.
> >>>
> >>> 1. CFI Plan Apo Lambda 100X Oil 1.45 NA (current lens)
> >>> 2. CFI Plan Apo Lambda DM 100X Oil 1.45 NA
> >>> 3. CFI Apo SR TIRF 100X Oil 1.49 NA
> >>>
> >>>
> >>> I think the Lambda lenses are supposed to be lower background, though
> I've
> >>> never tested whether that is important or not. They are also Plan
> lenses,
> >>> but I'm not sure how important that is for the <40 micron regions we
> >>> typically image. The SR lens has a correction collar which might be
> useful
> >>> (?) when imaging a bit off the glass. The online information says the
> SR
> >>> lens has a PSF optimized for STORM which sounds good but I'm not sure
> what
> >>> it actually means. I don't have any damage threshold information about
> >>> these
> >>> lenses. Oh, we are pretty sure the damage was caused when we sent ~120
> mW
> >>> at
> >>> 405 nm was sent to the lens in EPI and later in TIRF. We plan to
> >>> substantially attenuate the laser in the future, which didn't need to
> be
> >>> that big anyway, but I am not sure how much to attenuate by.
> >>>
> >>>
> >>> Joshua C. Vaughan
> >>> Assistant Professor
> >>> Department of Chemistry
> >>> Box 351700
> >>> University of Washington
> >>> Seattle, WA 98195
> >>>
> >>
> >> --
> >> Martin Wessendorf, Ph.D.                   office: [phone]
> >> Assoc Prof, Dept Neuroscience                 lab: [phone]
> >> University of Minnesota             Preferred FAX: [phone]
> >> 6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
> >> Minneapolis, MN  55455                    e-mail: [hidden email]
> >>
>

--
Joshua C. Vaughan
Assistant Professor
Department of Chemistry
Box 351700
University of Washington
Seattle, WA 98195
206-543-4644

[Craig Brideau]

*****
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.
*****

If 405nm caused a new bubble while 647nm didn't, that sounds like the glue
between the lenses is failing. The glues used in optics tend to be
sensitive to shorter wavelengths, and many are in fact UV-cured.

Craig

On Fri, May 27, 2016 at 10:53 AM, Joshua Vaughan <[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.
> *****
>
> Followup to some replies.
>
> Seamus:
> The various damage spots happened over a period of the last ~2 years. I
> have relatively large lasers, with 405/488/561/647/750 at
> ~200/1000/2000/1500/500 mW but we generally run at much lower powers. This
> is more laser power than most, I know, but I used something like this
> previously on an Olympus UPlanSApo 1.4 NA oil lens that never had trouble
> so I thought it would be ok. We noticed the damage first in the
> illumination since in 'straight epi' with the beam going up we got lots of
> scattering and uneven illumination so we got into the habit of aligning
> (with very low powers!) slightly off straight epi. Over time there was more
> damage, particularly near TIRF positions on either side of the lens. At
> this point, the PSF is noticeably asymmetric and starting to hurt things
> for 3D by astigmatism, though for 2D near focus things were ok-ish.
>
> Martin, Nuno, Craig:
> I can't say for sure whether the lens had gotten dirt or oil inside though
> I believe not. My group is generally careful, but I suppose someone could
> have gotten oil down the barrel of the Nikon lens (the Olympus UIS2
> mushroom-cap type design is better for keeping out oil). The image I posted
> has is focused roughly on the back focal plane so I believe the issue is
> interior to the objective. The back hemisphere of the lens looked fine to
> me when I examined reflections carefully near a bright light.
>
> Zdenek:
> Before sending the lens back, we did a couple tests in an initially
> undamaged region that showed no damage in the back focal plane. Lots of 647
> nm power (~1000 mW) did not lead to damage in the test, but lots of 405 nm
> power (~100 mW) led to a new and prominent 'bubble' being formed around the
> illuminated part of the BFP within 10-20 minutes. I figured if we were
> going to replace or repair we might as well try to figure out a little more
> information about the culprit so we can try to avoid this problem in the
> future.
>
> A little more info:
> Folks at my old postdoc lab tell me that they also see damage in their 60x
> version of the Lambda lens (I think it is CFI Plan Apo Lambda 60x Oil
> 1.4NA) although it is not as dramatic and that lens has been used for a
> shorter period than mine (~3 years of use). I wonder whether there are more
> damaged lenses out there that go unnoticed since it is a little hard to
> tell in TIRF and since I suspect few people look at the back focal plane.
> When I inspect, in addition to illuminating with the relatively low NA
> condenser like in the posted photo, sometimes I use epi-illumination of
> beads to fill the back aperture.
>
>
>
> On Fri, May 27, 2016 at 8:03 AM, Nuno Moreno <[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.
> > *****
> >
> > Dear Craig
> >
> > I sent already a message directly to Joshua with a similar statement. The
> > problem is however when the burning in not at the first lens but
> somewhere
> > inside. In this case this means that the objective was not assembled in a
> > proper clean environment and has dust or other particles inside. Actually
> > here at the institute we were surprised on how often this happens from 2
> > different manufacturers. Maybe we were just unlucky. If that is the case
> I
> > would ask for a replacement free of charge. I must confess however that
> in
> > Joshua case it does not look like but the picture is not clear enough. I
> > would advice you to bring your objective to a stereoscope, start to focus
> > on the first back optical element and check whether this is superficial
> or
> > not.
> >
> > Kind regards and good luck
> > Nuno Moreno
> >
> > > On 27 May 2016, at 15:07, Craig Brideau <[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.
> > > *****
> > >
> > > I had a glance at your picture. The black spots look like laser burns,
> > but
> > > there is also a clear bubble or edge that looks suspiciously like oil
> > also
> > > visible in the picture. It's possible you had some oil contamination
> and
> > > then it burned onto the glass. Most optics will handle a surprising
> > amount
> > > of laser power as long as they are extremely clean. The instant there
> is
> > > any dust/grease the laser will burn the surface as the contamination
> > > increases point absorption at the interface.
> > >
> > > Craig
> > >
> > > On Fri, May 27, 2016 at 7:28 AM, Martin Wessendorf <[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.
> > >> *****
> > >>
> > >> Dear Dr. Vaughan--
> > >>
> > >> Could it be dirt and oil on the lens, rather than being burned and/or
> > >> unglued?
> > >>
> > >> Martin Wessendorf
> > >>
> > >>
> > >>
> > >>
> > >> On 5/27/2016 2:58 AM, Joshua Vaughan 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 damaged an objective lens, a Nikon CFI Plan Apo Lambda 100X Oil
> > 1.45 NA
> > >>> lens on our homebuilt TIRF/STORM microscope. Has anybody seen this
> > sort of
> > >>> thing before? I'm interested to hear, if so. We will repair or
> replace
> > it,
> > >>> and if replacing I wonder what to get. More details are below.
> > >>>
> > >>> Here is a link to a pic of part of the back focal plane. It has some
> > dark
> > >>> spots that look 'burned' and some stuff that looks like 'bubbles'.
> > >>>
> > >>> http://i63.tinypic.com/mmemvl.jpg
> > >>>
> > >>>
> > >>> Repairing this lens will cost >50% of a new lens so I want to decide
> > >>> whether
> > >>> to repair or to just get something else. I'd prefer something with a
> > >>> higher
> > >>> damage threshold but I was unsuccessful in getting data on lens
> damage
> > >>> thresholds.
> > >>>
> > >>> If replacing, what to get? I noticed three lenses online and I wonder
> > what
> > >>> is best for STORM and whether all have a similar damage threshold or
> > not.
> > >>>
> > >>> 1. CFI Plan Apo Lambda 100X Oil 1.45 NA (current lens)
> > >>> 2. CFI Plan Apo Lambda DM 100X Oil 1.45 NA
> > >>> 3. CFI Apo SR TIRF 100X Oil 1.49 NA
> > >>>
> > >>>
> > >>> I think the Lambda lenses are supposed to be lower background, though
> > I've
> > >>> never tested whether that is important or not. They are also Plan
> > lenses,
> > >>> but I'm not sure how important that is for the <40 micron regions we
> > >>> typically image. The SR lens has a correction collar which might be
> > useful
> > >>> (?) when imaging a bit off the glass. The online information says the
> > SR
> > >>> lens has a PSF optimized for STORM which sounds good but I'm not sure
> > what
> > >>> it actually means. I don't have any damage threshold information
> about
> > >>> these
> > >>> lenses. Oh, we are pretty sure the damage was caused when we sent
> ~120
> > mW
> > >>> at
> > >>> 405 nm was sent to the lens in EPI and later in TIRF. We plan to
> > >>> substantially attenuate the laser in the future, which didn't need to
> > be
> > >>> that big anyway, but I am not sure how much to attenuate by.
> > >>>
> > >>>
> > >>> Joshua C. Vaughan
> > >>> Assistant Professor
> > >>> Department of Chemistry
> > >>> Box 351700
> > >>> University of Washington
> > >>> Seattle, WA 98195
> > >>>
> > >>
> > >> --
> > >> Martin Wessendorf, Ph.D.                   office: [phone]
> > >> Assoc Prof, Dept Neuroscience                 lab: [phone]
> > >> University of Minnesota             Preferred FAX: [phone]
> > >> 6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
> > >> Minneapolis, MN  55455                    e-mail: [hidden email]
> > >>
> >
>
>
>
> --
> Joshua C. Vaughan
> Assistant Professor
> Department of Chemistry
> Box 351700
> University of Washington
> Seattle, WA 98195
> 206-543-4644
>

[John Oreopoulos]

*****
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.
*****

Do you really need these high laser powers for your application?

John Oreopoulos

> On May 27, 2016, at 11:42 AM, Glen MacDonald <[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.
> *****
>
> Possibly the damage is to the optical cement between the lens elements.  I found epoxy to be very absorbtive and could be damaged during 2-photon imaging.  
>
> Regards,
> Glen MacDonald
> Digital Microscopy Center
> Box 357923
> University of Washington
> Seattle, WA 98195-7923  USA
> [phone]
> [hidden email]
>
>
>
>
>
>
>
>
>> On May 27, 2016, at 8:03 AM, Nuno Moreno <[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.
>> *****
>>
>> Dear Craig
>>
>> I sent already a message directly to Joshua with a similar statement. The problem is however when the burning in not at the first lens but somewhere inside. In this case this means that the objective was not assembled in a proper clean environment and has dust or other particles inside. Actually here at the institute we were surprised on how often this happens from 2 different manufacturers. Maybe we were just unlucky. If that is the case I would ask for a replacement free of charge. I must confess however that in Joshua case it does not look like but the picture is not clear enough. I would advice you to bring your objective to a stereoscope, start to focus on the first back optical element and check whether this is superficial or not.
>>
>> Kind regards and good luck
>> Nuno Moreno
>>
>>> On 27 May 2016, at 15:07, Craig Brideau <[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.
>>> *****
>>>
>>> I had a glance at your picture. The black spots look like laser burns, but
>>> there is also a clear bubble or edge that looks suspiciously like oil also
>>> visible in the picture. It's possible you had some oil contamination and
>>> then it burned onto the glass. Most optics will handle a surprising amount
>>> of laser power as long as they are extremely clean. The instant there is
>>> any dust/grease the laser will burn the surface as the contamination
>>> increases point absorption at the interface.
>>>
>>> Craig
>>>
>>>> On Fri, May 27, 2016 at 7:28 AM, Martin Wessendorf <[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.
>>>> *****
>>>>
>>>> Dear Dr. Vaughan--
>>>>
>>>> Could it be dirt and oil on the lens, rather than being burned and/or
>>>> unglued?
>>>>
>>>> Martin Wessendorf
>>>>
>>>>
>>>>
>>>>
>>>>> On 5/27/2016 2:58 AM, Joshua Vaughan 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 damaged an objective lens, a Nikon CFI Plan Apo Lambda 100X Oil 1.45 NA
>>>>> lens on our homebuilt TIRF/STORM microscope. Has anybody seen this sort of
>>>>> thing before? I'm interested to hear, if so. We will repair or replace it,
>>>>> and if replacing I wonder what to get. More details are below.
>>>>>
>>>>> Here is a link to a pic of part of the back focal plane. It has some dark
>>>>> spots that look 'burned' and some stuff that looks like 'bubbles'.
>>>>>
>>>>> http://i63.tinypic.com/mmemvl.jpg
>>>>>
>>>>>
>>>>> Repairing this lens will cost >50% of a new lens so I want to decide
>>>>> whether
>>>>> to repair or to just get something else. I'd prefer something with a
>>>>> higher
>>>>> damage threshold but I was unsuccessful in getting data on lens damage
>>>>> thresholds.
>>>>>
>>>>> If replacing, what to get? I noticed three lenses online and I wonder what
>>>>> is best for STORM and whether all have a similar damage threshold or not.
>>>>>
>>>>> 1. CFI Plan Apo Lambda 100X Oil 1.45 NA (current lens)
>>>>> 2. CFI Plan Apo Lambda DM 100X Oil 1.45 NA
>>>>> 3. CFI Apo SR TIRF 100X Oil 1.49 NA
>>>>>
>>>>>
>>>>> I think the Lambda lenses are supposed to be lower background, though I've
>>>>> never tested whether that is important or not. They are also Plan lenses,
>>>>> but I'm not sure how important that is for the <40 micron regions we
>>>>> typically image. The SR lens has a correction collar which might be useful
>>>>> (?) when imaging a bit off the glass. The online information says the SR
>>>>> lens has a PSF optimized for STORM which sounds good but I'm not sure what
>>>>> it actually means. I don't have any damage threshold information about
>>>>> these
>>>>> lenses. Oh, we are pretty sure the damage was caused when we sent ~120 mW
>>>>> at
>>>>> 405 nm was sent to the lens in EPI and later in TIRF. We plan to
>>>>> substantially attenuate the laser in the future, which didn't need to be
>>>>> that big anyway, but I am not sure how much to attenuate by.
>>>>>
>>>>>
>>>>> Joshua C. Vaughan
>>>>> Assistant Professor
>>>>> Department of Chemistry
>>>>> Box 351700
>>>>> University of Washington
>>>>> Seattle, WA 98195
>>>>
>>>> --
>>>> Martin Wessendorf, Ph.D.                   office: [phone]
>>>> Assoc Prof, Dept Neuroscience                 lab: [phone]
>>>> University of Minnesota             Preferred FAX: [phone]
>>>> 6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
>>>> Minneapolis, MN  55455                    e-mail: [hidden email]
>>>>

[Sripad Ram-2]

*****
To join, leave or search the confocal microscopy listserv, go to:
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Post images on http://www.imgur.com and include the link in your posting.
*****

The bubbles is something that I observed years ago (postdoc days) even with
dry lenses (20x PApo; Zeiss) that has never been illuminated with a laser.
At that time, we suspected that someone accidentally used oil on the dry
lens and forgot to wipe it off and was never noticed for a very long time.
It is possible that it could well be that something internal was damaged.

Sripad


On Fri, May 27, 2016 at 10:41 AM, John Oreopoulos <
[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.
> *****
>
> Do you really need these high laser powers for your application?
>
> John Oreopoulos
>
> > On May 27, 2016, at 11:42 AM, Glen MacDonald <[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.
> > *****
> >
> > Possibly the damage is to the optical cement between the lens elements.
> I found epoxy to be very absorbtive and could be damaged during 2-photon
> imaging.
> >
> > Regards,
> > Glen MacDonald
> > Digital Microscopy Center
> > Box 357923
> > University of Washington
> > Seattle, WA 98195-7923  USA
> > [phone]
> > [hidden email]
> >
> >
> >
> >
> >
> >
> >
> >
> >> On May 27, 2016, at 8:03 AM, Nuno Moreno <[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.
> >> *****
> >>
> >> Dear Craig
> >>
> >> I sent already a message directly to Joshua with a similar statement.
> The problem is however when the burning in not at the first lens but
> somewhere inside. In this case this means that the objective was not
> assembled in a proper clean environment and has dust or other particles
> inside. Actually here at the institute we were surprised on how often this
> happens from 2 different manufacturers. Maybe we were just unlucky. If that
> is the case I would ask for a replacement free of charge. I must confess
> however that in Joshua case it does not look like but the picture is not
> clear enough. I would advice you to bring your objective to a stereoscope,
> start to focus on the first back optical element and check whether this is
> superficial or not.
> >>
> >> Kind regards and good luck
> >> Nuno Moreno
> >>
> >>> On 27 May 2016, at 15:07, Craig Brideau <[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.
> >>> *****
> >>>
> >>> I had a glance at your picture. The black spots look like laser burns,
> but
> >>> there is also a clear bubble or edge that looks suspiciously like oil
> also
> >>> visible in the picture. It's possible you had some oil contamination
> and
> >>> then it burned onto the glass. Most optics will handle a surprising
> amount
> >>> of laser power as long as they are extremely clean. The instant there
> is
> >>> any dust/grease the laser will burn the surface as the contamination
> >>> increases point absorption at the interface.
> >>>
> >>> Craig
> >>>
> >>>> On Fri, May 27, 2016 at 7:28 AM, Martin Wessendorf <[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.
> >>>> *****
> >>>>
> >>>> Dear Dr. Vaughan--
> >>>>
> >>>> Could it be dirt and oil on the lens, rather than being burned and/or
> >>>> unglued?
> >>>>
> >>>> Martin Wessendorf
> >>>>
> >>>>
> >>>>
> >>>>
> >>>>> On 5/27/2016 2:58 AM, Joshua Vaughan 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 damaged an objective lens, a Nikon CFI Plan Apo Lambda 100X Oil
> 1.45 NA
> >>>>> lens on our homebuilt TIRF/STORM microscope. Has anybody seen this
> sort of
> >>>>> thing before? I'm interested to hear, if so. We will repair or
> replace it,
> >>>>> and if replacing I wonder what to get. More details are below.
> >>>>>
> >>>>> Here is a link to a pic of part of the back focal plane. It has some
> dark
> >>>>> spots that look 'burned' and some stuff that looks like 'bubbles'.
> >>>>>
> >>>>> http://i63.tinypic.com/mmemvl.jpg
> >>>>>
> >>>>>
> >>>>> Repairing this lens will cost >50% of a new lens so I want to decide
> >>>>> whether
> >>>>> to repair or to just get something else. I'd prefer something with a
> >>>>> higher
> >>>>> damage threshold but I was unsuccessful in getting data on lens
> damage
> >>>>> thresholds.
> >>>>>
> >>>>> If replacing, what to get? I noticed three lenses online and I
> wonder what
> >>>>> is best for STORM and whether all have a similar damage threshold or
> not.
> >>>>>
> >>>>> 1. CFI Plan Apo Lambda 100X Oil 1.45 NA (current lens)
> >>>>> 2. CFI Plan Apo Lambda DM 100X Oil 1.45 NA
> >>>>> 3. CFI Apo SR TIRF 100X Oil 1.49 NA
> >>>>>
> >>>>>
> >>>>> I think the Lambda lenses are supposed to be lower background,
> though I've
> >>>>> never tested whether that is important or not. They are also Plan
> lenses,
> >>>>> but I'm not sure how important that is for the <40 micron regions we
> >>>>> typically image. The SR lens has a correction collar which might be
> useful
> >>>>> (?) when imaging a bit off the glass. The online information says
> the SR
> >>>>> lens has a PSF optimized for STORM which sounds good but I'm not
> sure what
> >>>>> it actually means. I don't have any damage threshold information
> about
> >>>>> these
> >>>>> lenses. Oh, we are pretty sure the damage was caused when we sent
> ~120 mW
> >>>>> at
> >>>>> 405 nm was sent to the lens in EPI and later in TIRF. We plan to
> >>>>> substantially attenuate the laser in the future, which didn't need
> to be
> >>>>> that big anyway, but I am not sure how much to attenuate by.
> >>>>>
> >>>>>
> >>>>> Joshua C. Vaughan
> >>>>> Assistant Professor
> >>>>> Department of Chemistry
> >>>>> Box 351700
> >>>>> University of Washington
> >>>>> Seattle, WA 98195
> >>>>
> >>>> --
> >>>> Martin Wessendorf, Ph.D.                   office: [phone]
> >>>> Assoc Prof, Dept Neuroscience                 lab: [phone]
> >>>> University of Minnesota             Preferred FAX: [phone]
> >>>> 6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
> >>>> Minneapolis, MN  55455                    e-mail: [hidden email]
> >>>>
>

*****
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.
*****

Thanks for several thoughtful replies. I have a couple of follow up points.

Is anyone aware of damage threshold data for objective lenses, from Nikon
or other manufacturers? It seems many people suspect glue damage for my
lens. I asked my local Nikon sales rep but he was unable to get data for
me.

Does anybody have a favorite objective lens for STORM/PALM among the
following Nikon lenses? I'm intrigued by #4, below, which is rated for high
power (Seamus mentioned).

1. CFI Plan Apo Lambda 100X Oil 1.45 NA (current lens)
2. CFI Plan Apo Lambda DM 100X Oil 1.45 NA
3. CFI Apo SR TIRF 100X Oil 1.49 NA
4. CFI HP Apochromat TIRF 100x oil 1.49 NA


To John Oreopoulos... No, we don't need 1W lasers for imaging. We generally
work with up to 50-100 mW for the 488/561/647/750 nm lines and use the rest
of the laser power for other stuff including spectroscopy experiments. For
the 405 nm line, it is typically below 1 mW, but considering our test which
showed 405 nm is likely the problem, I wonder if a user may have blasted
with 405 nm at some point leading to damage that got worse with repeated
use even at lower power.

On Fri, May 27, 2016 at 2:31 PM, S Ram <[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.
> *****
>
> The bubbles is something that I observed years ago (postdoc days) even with
> dry lenses (20x PApo; Zeiss) that has never been illuminated with a laser.
> At that time, we suspected that someone accidentally used oil on the dry
> lens and forgot to wipe it off and was never noticed for a very long time.
> It is possible that it could well be that something internal was damaged.
>
> Sripad
>
>
> On Fri, May 27, 2016 at 10:41 AM, John Oreopoulos <
> [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.
> > *****
> >
> > Do you really need these high laser powers for your application?
> >
> > John Oreopoulos
> >
> > > On May 27, 2016, at 11:42 AM, Glen MacDonald <[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.
> > > *****
> > >
> > > Possibly the damage is to the optical cement between the lens elements.
> > I found epoxy to be very absorbtive and could be damaged during 2-photon
> > imaging.
> > >
> > > Regards,
> > > Glen MacDonald
> > > Digital Microscopy Center
> > > Box 357923
> > > University of Washington
> > > Seattle, WA 98195-7923  USA
> > > [phone]
> > > [hidden email]
> > >
> > >
> > >
> > >
> > >
> > >
> > >
> > >
> > >> On May 27, 2016, at 8:03 AM, Nuno Moreno <[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.
> > >> *****
> > >>
> > >> Dear Craig
> > >>
> > >> I sent already a message directly to Joshua with a similar statement.
> > The problem is however when the burning in not at the first lens but
> > somewhere inside. In this case this means that the objective was not
> > assembled in a proper clean environment and has dust or other particles
> > inside. Actually here at the institute we were surprised on how often
> this
> > happens from 2 different manufacturers. Maybe we were just unlucky. If
> that
> > is the case I would ask for a replacement free of charge. I must confess
> > however that in Joshua case it does not look like but the picture is not
> > clear enough. I would advice you to bring your objective to a
> stereoscope,
> > start to focus on the first back optical element and check whether this
> is
> > superficial or not.
> > >>
> > >> Kind regards and good luck
> > >> Nuno Moreno
> > >>
> > >>> On 27 May 2016, at 15:07, Craig Brideau <[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.
> > >>> *****
> > >>>
> > >>> I had a glance at your picture. The black spots look like laser
> burns,
> > but
> > >>> there is also a clear bubble or edge that looks suspiciously like oil
> > also
> > >>> visible in the picture. It's possible you had some oil contamination
> > and
> > >>> then it burned onto the glass. Most optics will handle a surprising
> > amount
> > >>> of laser power as long as they are extremely clean. The instant there
> > is
> > >>> any dust/grease the laser will burn the surface as the contamination
> > >>> increases point absorption at the interface.
> > >>>
> > >>> Craig
> > >>>
> > >>>> On Fri, May 27, 2016 at 7:28 AM, Martin Wessendorf <[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.
> > >>>> *****
> > >>>>
> > >>>> Dear Dr. Vaughan--
> > >>>>
> > >>>> Could it be dirt and oil on the lens, rather than being burned
> and/or
> > >>>> unglued?
> > >>>>
> > >>>> Martin Wessendorf
> > >>>>
> > >>>>
> > >>>>
> > >>>>
> > >>>>> On 5/27/2016 2:58 AM, Joshua Vaughan 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 damaged an objective lens, a Nikon CFI Plan Apo Lambda 100X Oil
> > 1.45 NA
> > >>>>> lens on our homebuilt TIRF/STORM microscope. Has anybody seen this
> > sort of
> > >>>>> thing before? I'm interested to hear, if so. We will repair or
> > replace it,
> > >>>>> and if replacing I wonder what to get. More details are below.
> > >>>>>
> > >>>>> Here is a link to a pic of part of the back focal plane. It has
> some
> > dark
> > >>>>> spots that look 'burned' and some stuff that looks like 'bubbles'.
> > >>>>>
> > >>>>> http://i63.tinypic.com/mmemvl.jpg
> > >>>>>
> > >>>>>
> > >>>>> Repairing this lens will cost >50% of a new lens so I want to
> decide
> > >>>>> whether
> > >>>>> to repair or to just get something else. I'd prefer something with
> a
> > >>>>> higher
> > >>>>> damage threshold but I was unsuccessful in getting data on lens
> > damage
> > >>>>> thresholds.
> > >>>>>
> > >>>>> If replacing, what to get? I noticed three lenses online and I
> > wonder what
> > >>>>> is best for STORM and whether all have a similar damage threshold
> or
> > not.
> > >>>>>
> > >>>>> 1. CFI Plan Apo Lambda 100X Oil 1.45 NA (current lens)
> > >>>>> 2. CFI Plan Apo Lambda DM 100X Oil 1.45 NA
> > >>>>> 3. CFI Apo SR TIRF 100X Oil 1.49 NA
> > >>>>>
> > >>>>>
> > >>>>> I think the Lambda lenses are supposed to be lower background,
> > though I've
> > >>>>> never tested whether that is important or not. They are also Plan
> > lenses,
> > >>>>> but I'm not sure how important that is for the <40 micron regions
> we
> > >>>>> typically image. The SR lens has a correction collar which might be
> > useful
> > >>>>> (?) when imaging a bit off the glass. The online information says
> > the SR
> > >>>>> lens has a PSF optimized for STORM which sounds good but I'm not
> > sure what
> > >>>>> it actually means. I don't have any damage threshold information
> > about
> > >>>>> these
> > >>>>> lenses. Oh, we are pretty sure the damage was caused when we sent
> > ~120 mW
> > >>>>> at
> > >>>>> 405 nm was sent to the lens in EPI and later in TIRF. We plan to
> > >>>>> substantially attenuate the laser in the future, which didn't need
> > to be
> > >>>>> that big anyway, but I am not sure how much to attenuate by.
> > >>>>>
> > >>>>>
> > >>>>> Joshua C. Vaughan
> > >>>>> Assistant Professor
> > >>>>> Department of Chemistry
> > >>>>> Box 351700
> > >>>>> University of Washington
> > >>>>> Seattle, WA 98195
> > >>>>
> > >>>> --
> > >>>> Martin Wessendorf, Ph.D.                   office: [phone]
> > >>>> Assoc Prof, Dept Neuroscience                 lab: [phone]
> > >>>> University of Minnesota             Preferred FAX: [phone]
> > >>>> 6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
> > >>>> Minneapolis, MN  55455                    e-mail: [hidden email]
> > >>>>
> >
>

--
Joshua C. Vaughan
Assistant Professor
Department of Chemistry
Box 351700
University of Washington
Seattle, WA 98195
206-543-4644

[Andreas Bruckbauer]

*****
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 Joshua,


several years ago, i discussed the maximum laser power one could focus at the back focal plane of an objective with an optical engineer from Olympus. At that time they just had increased the laser power of their TIRF lasers from 100 mW to 200 mW for 488 nm and that was regarded as save for their TIRF objectives (this is usually the power before coupling into the optical fibre and you expect 70% of that . We discussed the case of the OMX microscope, which uses Olympus objectives and higher laser powers (around 300 - 500mW) and he was concerned about possible damage, mainly of the coating. However in this case I think the laser is not focused on the BF plane. The Bruker Vutara microscope uses Olympus lenses and powers of 1000 mW for 488, 561, 642 nm but "only" 100 mW at 405 nm. But as i understand this is usually set up as a WF microscope for 3D imaging, so they also do not need to focus to a point. It is also interesting to look at the objective transmission data when available. The Olympus 100x TIRF objective e.g. has > 80% transmission from 400 - 700nm, while the Zeiss 100x TIRF lens starts with only 50% at 400 nm and stays below 70% for the visible spectrum. Unfortunately i could not find the data for Nikon or Leica objectives.


best wishes


Andreas










-----Original Message-----
From: Joshua Vaughan <[hidden email]>
To: CONFOCALMICROSCOPY <[hidden email]>
Sent: Tue, 31 May 2016 1:30
Subject: Re: objective lens laser damage

*****
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.
*****

Thanks for several thoughtful replies. I have a couple of follow up points.

Is anyone aware of damage threshold data for objective lenses, from Nikon
or other manufacturers? It seems many people suspect glue damage for my
lens. I asked my local Nikon sales rep but he was unable to get data for
me.

Does anybody have a favorite objective lens for STORM/PALM among the
following Nikon lenses? I'm intrigued by #4, below, which is rated for high
power (Seamus mentioned).

1. CFI Plan Apo Lambda 100X Oil 1.45 NA (current lens)
2. CFI Plan Apo Lambda DM 100X Oil 1.45 NA
3. CFI Apo SR TIRF 100X Oil 1.49 NA
4. CFI HP Apochromat TIRF 100x oil 1.49 NA


To John Oreopoulos... No, we don't need 1W lasers for imaging. We generally
work with up to 50-100 mW for the 488/561/647/750 nm lines and use the rest
of the laser power for other stuff including spectroscopy experiments. For
the 405 nm line, it is typically below 1 mW, but considering our test which
showed 405 nm is likely the problem, I wonder if a user may have blasted
with 405 nm at some point leading to damage that got worse with repeated
use even at lower power.

On Fri, May 27, 2016 at 2:31 PM, S Ram <[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.
> *****
>
> The bubbles is something that I observed years ago (postdoc days) even with
> dry lenses (20x PApo; Zeiss) that has never been illuminated with a laser.
> At that time, we suspected that someone accidentally used oil on the dry
> lens and forgot to wipe it off and was never noticed for a very long time.
> It is possible that it could well be that something internal was damaged.
>
> Sripad
>
>
> On Fri, May 27, 2016 at 10:41 AM, John Oreopoulos <
> [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.
> > *****
> >
> > Do you really need these high laser powers for your application?
> >
> > John Oreopoulos
> >
> > > On May 27, 2016, at 11:42 AM, Glen MacDonald <[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.
> > > *****
> > >
> > > Possibly the damage is to the optical cement between the lens elements.
> > I found epoxy to be very absorbtive and could be damaged during 2-photon
> > imaging.
> > >
> > > Regards,
> > > Glen MacDonald
> > > Digital Microscopy Center
> > > Box 357923
> > > University of Washington
> > > Seattle, WA 98195-7923  USA
> > > [phone]
> > > [hidden email]
> > >
> > >
> > >
> > >
> > >
> > >
> > >
> > >
> > >> On May 27, 2016, at 8:03 AM, Nuno Moreno <[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.
> > >> *****
> > >>
> > >> Dear Craig
> > >>
> > >> I sent already a message directly to Joshua with a similar statement.
> > The problem is however when the burning in not at the first lens but
> > somewhere inside. In this case this means that the objective was not
> > assembled in a proper clean environment and has dust or other particles
> > inside. Actually here at the institute we were surprised on how often
> this
> > happens from 2 different manufacturers. Maybe we were just unlucky. If
> that
> > is the case I would ask for a replacement free of charge. I must confess
> > however that in Joshua case it does not look like but the picture is not
> > clear enough. I would advice you to bring your objective to a
> stereoscope,
> > start to focus on the first back optical element and check whether this
> is
> > superficial or not.
> > >>
> > >> Kind regards and good luck
> > >> Nuno Moreno
> > >>
> > >>> On 27 May 2016, at 15:07, Craig Brideau <[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.
> > >>> *****
> > >>>
> > >>> I had a glance at your picture. The black spots look like laser
> burns,
> > but
> > >>> there is also a clear bubble or edge that looks suspiciously like oil
> > also
> > >>> visible in the picture. It's possible you had some oil contamination
> > and
> > >>> then it burned onto the glass. Most optics will handle a surprising
> > amount
> > >>> of laser power as long as they are extremely clean. The instant there
> > is
> > >>> any dust/grease the laser will burn the surface as the contamination
> > >>> increases point absorption at the interface.
> > >>>
> > >>> Craig
> > >>>
> > >>>> On Fri, May 27, 2016 at 7:28 AM, Martin Wessendorf <[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.
> > >>>> *****
> > >>>>
> > >>>> Dear Dr. Vaughan--
> > >>>>
> > >>>> Could it be dirt and oil on the lens, rather than being burned
> and/or
> > >>>> unglued?
> > >>>>
> > >>>> Martin Wessendorf
> > >>>>
> > >>>>
> > >>>>
> > >>>>
> > >>>>> On 5/27/2016 2:58 AM, Joshua Vaughan 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 damaged an objective lens, a Nikon CFI Plan Apo Lambda 100X Oil
> > 1.45 NA
> > >>>>> lens on our homebuilt TIRF/STORM microscope. Has anybody seen this
> > sort of
> > >>>>> thing before? I'm interested to hear, if so. We will repair or
> > replace it,
> > >>>>> and if replacing I wonder what to get. More details are below.
> > >>>>>
> > >>>>> Here is a link to a pic of part of the back focal plane. It has
> some
> > dark
> > >>>>> spots that look 'burned' and some stuff that looks like 'bubbles'.
> > >>>>>
> > >>>>> http://i63.tinypic.com/mmemvl.jpg
> > >>>>>
> > >>>>>
> > >>>>> Repairing this lens will cost >50% of a new lens so I want to
> decide
> > >>>>> whether
> > >>>>> to repair or to just get something else. I'd prefer something with
> a
> > >>>>> higher
> > >>>>> damage threshold but I was unsuccessful in getting data on lens
> > damage
> > >>>>> thresholds.
> > >>>>>
> > >>>>> If replacing, what to get? I noticed three lenses online and I
> > wonder what
> > >>>>> is best for STORM and whether all have a similar damage threshold
> or
> > not.
> > >>>>>
> > >>>>> 1. CFI Plan Apo Lambda 100X Oil 1.45 NA (current lens)
> > >>>>> 2. CFI Plan Apo Lambda DM 100X Oil 1.45 NA
> > >>>>> 3. CFI Apo SR TIRF 100X Oil 1.49 NA
> > >>>>>
> > >>>>>
> > >>>>> I think the Lambda lenses are supposed to be lower background,
> > though I've
> > >>>>> never tested whether that is important or not. They are also Plan
> > lenses,
> > >>>>> but I'm not sure how important that is for the <40 micron regions
> we
> > >>>>> typically image. The SR lens has a correction collar which might be
> > useful
> > >>>>> (?) when imaging a bit off the glass. The online information says
> > the SR
> > >>>>> lens has a PSF optimized for STORM which sounds good but I'm not
> > sure what
> > >>>>> it actually means. I don't have any damage threshold information
> > about
> > >>>>> these
> > >>>>> lenses. Oh, we are pretty sure the damage was caused when we sent
> > ~120 mW
> > >>>>> at
> > >>>>> 405 nm was sent to the lens in EPI and later in TIRF. We plan to
> > >>>>> substantially attenuate the laser in the future, which didn't need
> > to be
> > >>>>> that big anyway, but I am not sure how much to attenuate by.
> > >>>>>
> > >>>>>
> > >>>>> Joshua C. Vaughan
> > >>>>> Assistant Professor
> > >>>>> Department of Chemistry
> > >>>>> Box 351700
> > >>>>> University of Washington
> > >>>>> Seattle, WA 98195
> > >>>>
> > >>>> --
> > >>>> Martin Wessendorf, Ph.D.                   office: [phone]
> > >>>> Assoc Prof, Dept Neuroscience                 lab: [phone]
> > >>>> University of Minnesota             Preferred FAX: [phone]
> > >>>> 6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
> > >>>> Minneapolis, MN  55455                    e-mail: [hidden email]
> > >>>>
> >
>

--
Joshua C. Vaughan
Assistant Professor
Department of Chemistry
Box 351700
University of Washington
Seattle, WA 98195
206-543-4644

[Bram van den Broek]

Dear Joshua and others,

Does it have to be a Nikon lens? Leica has a special GSD/dSTORM high power objective (160X Oil/1.43 NA) that supposedly doesn't have cement in between the optical elements.
We have been using it practically every day for several years now in their SR-GSD system, equipped with 300-500 mW lasers (488, 532 and 642 nm) and ~50 mW (405 nm) in TIRF and EPI, without any visible damage.

Best regards,
Kees Jalink & Bram van den Broek




-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Joshua Vaughan
Sent: dinsdag 31 mei 2016 2:26
To: [hidden email]
Subject: Re: objective lens laser damage

*****
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.
*****

Thanks for several thoughtful replies. I have a couple of follow up points.

Is anyone aware of damage threshold data for objective lenses, from Nikon or other manufacturers? It seems many people suspect glue damage for my lens. I asked my local Nikon sales rep but he was unable to get data for me.

Does anybody have a favorite objective lens for STORM/PALM among the following Nikon lenses? I'm intrigued by #4, below, which is rated for high power (Seamus mentioned).

1. CFI Plan Apo Lambda 100X Oil 1.45 NA (current lens) 2. CFI Plan Apo Lambda DM 100X Oil 1.45 NA 3. CFI Apo SR TIRF 100X Oil 1.49 NA 4. CFI HP Apochromat TIRF 100x oil 1.49 NA


To John Oreopoulos... No, we don't need 1W lasers for imaging. We generally work with up to 50-100 mW for the 488/561/647/750 nm lines and use the rest of the laser power for other stuff including spectroscopy experiments. For the 405 nm line, it is typically below 1 mW, but considering our test which showed 405 nm is likely the problem, I wonder if a user may have blasted with 405 nm at some point leading to damage that got worse with repeated use even at lower power.

On Fri, May 27, 2016 at 2:31 PM, S Ram <[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.
> *****
>
> The bubbles is something that I observed years ago (postdoc days) even
> with dry lenses (20x PApo; Zeiss) that has never been illuminated with a laser.
> At that time, we suspected that someone accidentally used oil on the
> dry lens and forgot to wipe it off and was never noticed for a very long time.
> It is possible that it could well be that something internal was damaged.
>
> Sripad
>
>
> On Fri, May 27, 2016 at 10:41 AM, John Oreopoulos <
> [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.
> > *****
> >
> > Do you really need these high laser powers for your application?
> >
> > John Oreopoulos
> >
> > > On May 27, 2016, at 11:42 AM, Glen MacDonald
> > > <[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.
> > > *****
> > >
> > > Possibly the damage is to the optical cement between the lens elements.
> > I found epoxy to be very absorbtive and could be damaged during
> > 2-photon imaging.
> > >
> > > Regards,
> > > Glen MacDonald
> > > Digital Microscopy Center
> > > Box 357923
> > > University of Washington
> > > Seattle, WA 98195-7923  USA
> > > [phone]
> > > [hidden email]
> > >
> > >
> > >
> > >
> > >
> > >
> > >
> > >
> > >> On May 27, 2016, at 8:03 AM, Nuno Moreno
> > >> <[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.
> > >> *****
> > >>
> > >> Dear Craig
> > >>
> > >> I sent already a message directly to Joshua with a similar statement.
> > The problem is however when the burning in not at the first lens but
> > somewhere inside. In this case this means that the objective was not
> > assembled in a proper clean environment and has dust or other
> > particles inside. Actually here at the institute we were surprised
> > on how often
> this
> > happens from 2 different manufacturers. Maybe we were just unlucky.
> > If
> that
> > is the case I would ask for a replacement free of charge. I must
> > confess however that in Joshua case it does not look like but the
> > picture is not clear enough. I would advice you to bring your
> > objective to a
> stereoscope,
> > start to focus on the first back optical element and check whether
> > this
> is
> > superficial or not.
> > >>
> > >> Kind regards and good luck
> > >> Nuno Moreno
> > >>
> > >>> On 27 May 2016, at 15:07, Craig Brideau
> > >>> <[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.
> > >>> *****
> > >>>
> > >>> I had a glance at your picture. The black spots look like laser
> burns,
> > but
> > >>> there is also a clear bubble or edge that looks suspiciously
> > >>> like oil
> > also
> > >>> visible in the picture. It's possible you had some oil
> > >>> contamination
> > and
> > >>> then it burned onto the glass. Most optics will handle a
> > >>> surprising
> > amount
> > >>> of laser power as long as they are extremely clean. The instant
> > >>> there
> > is
> > >>> any dust/grease the laser will burn the surface as the
> > >>> contamination increases point absorption at the interface.
> > >>>
> > >>> Craig
> > >>>
> > >>>> On Fri, May 27, 2016 at 7:28 AM, Martin Wessendorf
> > >>>> <[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.
> > >>>> *****
> > >>>>
> > >>>> Dear Dr. Vaughan--
> > >>>>
> > >>>> Could it be dirt and oil on the lens, rather than being burned
> and/or
> > >>>> unglued?
> > >>>>
> > >>>> Martin Wessendorf
> > >>>>
> > >>>>
> > >>>>
> > >>>>
> > >>>>> On 5/27/2016 2:58 AM, Joshua Vaughan 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 damaged an objective lens, a Nikon CFI Plan Apo Lambda 100X
> > >>>>> Oil
> > 1.45 NA
> > >>>>> lens on our homebuilt TIRF/STORM microscope. Has anybody seen
> > >>>>> this
> > sort of
> > >>>>> thing before? I'm interested to hear, if so. We will repair or
> > replace it,
> > >>>>> and if replacing I wonder what to get. More details are below.
> > >>>>>
> > >>>>> Here is a link to a pic of part of the back focal plane. It
> > >>>>> has
> some
> > dark
> > >>>>> spots that look 'burned' and some stuff that looks like 'bubbles'.
> > >>>>>
> > >>>>> http://i63.tinypic.com/mmemvl.jpg
> > >>>>>
> > >>>>>
> > >>>>> Repairing this lens will cost >50% of a new lens so I want to
> decide
> > >>>>> whether
> > >>>>> to repair or to just get something else. I'd prefer something
> > >>>>> with
> a
> > >>>>> higher
> > >>>>> damage threshold but I was unsuccessful in getting data on
> > >>>>> lens
> > damage
> > >>>>> thresholds.
> > >>>>>
> > >>>>> If replacing, what to get? I noticed three lenses online and I
> > wonder what
> > >>>>> is best for STORM and whether all have a similar damage
> > >>>>> threshold
> or
> > not.
> > >>>>>
> > >>>>> 1. CFI Plan Apo Lambda 100X Oil 1.45 NA (current lens) 2. CFI
> > >>>>> Plan Apo Lambda DM 100X Oil 1.45 NA 3. CFI Apo SR TIRF 100X
> > >>>>> Oil 1.49 NA
> > >>>>>
> > >>>>>
> > >>>>> I think the Lambda lenses are supposed to be lower background,
> > though I've
> > >>>>> never tested whether that is important or not. They are also
> > >>>>> Plan
> > lenses,
> > >>>>> but I'm not sure how important that is for the <40 micron
> > >>>>> regions
> we
> > >>>>> typically image. The SR lens has a correction collar which
> > >>>>> might be
> > useful
> > >>>>> (?) when imaging a bit off the glass. The online information
> > >>>>> says
> > the SR
> > >>>>> lens has a PSF optimized for STORM which sounds good but I'm
> > >>>>> not
> > sure what
> > >>>>> it actually means. I don't have any damage threshold
> > >>>>> information
> > about
> > >>>>> these
> > >>>>> lenses. Oh, we are pretty sure the damage was caused when we
> > >>>>> sent
> > ~120 mW
> > >>>>> at
> > >>>>> 405 nm was sent to the lens in EPI and later in TIRF. We plan
> > >>>>> to substantially attenuate the laser in the future, which
> > >>>>> didn't need
> > to be
> > >>>>> that big anyway, but I am not sure how much to attenuate by.
> > >>>>>
> > >>>>>
> > >>>>> Joshua C. Vaughan
> > >>>>> Assistant Professor
> > >>>>> Department of Chemistry
> > >>>>> Box 351700
> > >>>>> University of Washington
> > >>>>> Seattle, WA 98195
> > >>>>
> > >>>> --
> > >>>> Martin Wessendorf, Ph.D.                   office: [phone]
> > >>>> Assoc Prof, Dept Neuroscience                 lab: [phone]
> > >>>> University of Minnesota             Preferred FAX: [phone]
> > >>>> 6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
> > >>>> Minneapolis, MN  55455                    e-mail: [hidden email]
> > >>>>
> >
>

--
Joshua C. Vaughan
Assistant Professor
Department of Chemistry
Box 351700
University of Washington
Seattle, WA 98195
206-543-4644

[Reece, Jeff (NIH/NIDDK) [E]]

I thought Zeiss might say something, but I believe the latest 100x TIRF lens will have similar %T to the Olympus.
You can compare a few Zeiss TIRF lenses here:
https://www.micro-shop.zeiss.com/?s=306493725d39c2&l=en&p=us&f=o&a=c&m=s&pid[]=420780-9970-000&pid[]=420792-9800-000&pid[]=420792-9800-720&o=&st=1

Cheers,
Jeff

-----Original Message-----
From: Andreas Bruckbauer [mailto:[hidden email]]
Sent: Tuesday, May 31, 2016 3:56 AM
To: [hidden email]
Subject: Re: objective lens laser damage

*****
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 Joshua,


several years ago, i discussed the maximum laser power one could focus at the back focal plane of an objective with an optical engineer from Olympus. At that time they just had increased the laser power of their TIRF lasers from 100 mW to 200 mW for 488 nm and that was regarded as save for their TIRF objectives (this is usually the power before coupling into the optical fibre and you expect 70% of that . We discussed the case of the OMX microscope, which uses Olympus objectives and higher laser powers (around 300 - 500mW) and he was concerned about possible damage, mainly of the coating. However in this case I think the laser is not focused on the BF plane. The Bruker Vutara microscope uses Olympus lenses and powers of 1000 mW for 488, 561, 642 nm but "only" 100 mW at 405 nm. But as i understand this is usually set up as a WF microscope for 3D imaging, so they also do not need to focus to a point. It is also interesting to look at the objective transmission data when available. The Olympus 100x TIRF objective e.g. has > 80% transmission from 400 - 700nm, while the Zeiss 100x TIRF lens starts with only 50% at 400 nm and stays below 70% for the visible spectrum. Unfortunately i could not find the data for Nikon or Leica objectives.


best wishes


Andreas










-----Original Message-----
From: Joshua Vaughan <[hidden email]>
To: CONFOCALMICROSCOPY <[hidden email]>
Sent: Tue, 31 May 2016 1:30
Subject: Re: objective lens laser damage

*****
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.
*****

Thanks for several thoughtful replies. I have a couple of follow up points.

Is anyone aware of damage threshold data for objective lenses, from Nikon or other manufacturers? It seems many people suspect glue damage for my lens. I asked my local Nikon sales rep but he was unable to get data for me.

Does anybody have a favorite objective lens for STORM/PALM among the following Nikon lenses? I'm intrigued by #4, below, which is rated for high power (Seamus mentioned).

1. CFI Plan Apo Lambda 100X Oil 1.45 NA (current lens) 2. CFI Plan Apo Lambda DM 100X Oil 1.45 NA 3. CFI Apo SR TIRF 100X Oil 1.49 NA 4. CFI HP Apochromat TIRF 100x oil 1.49 NA


To John Oreopoulos... No, we don't need 1W lasers for imaging. We generally work with up to 50-100 mW for the 488/561/647/750 nm lines and use the rest of the laser power for other stuff including spectroscopy experiments. For the 405 nm line, it is typically below 1 mW, but considering our test which showed 405 nm is likely the problem, I wonder if a user may have blasted with 405 nm at some point leading to damage that got worse with repeated use even at lower power.

On Fri, May 27, 2016 at 2:31 PM, S Ram <[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.
> *****
>
> The bubbles is something that I observed years ago (postdoc days) even
> with dry lenses (20x PApo; Zeiss) that has never been illuminated with a laser.
> At that time, we suspected that someone accidentally used oil on the
> dry lens and forgot to wipe it off and was never noticed for a very long time.
> It is possible that it could well be that something internal was damaged.
>
> Sripad
>
>
> On Fri, May 27, 2016 at 10:41 AM, John Oreopoulos <
> [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.
> > *****
> >
> > Do you really need these high laser powers for your application?
> >
> > John Oreopoulos
> >
> > > On May 27, 2016, at 11:42 AM, Glen MacDonald
> > > <[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.
> > > *****
> > >
> > > Possibly the damage is to the optical cement between the lens elements.
> > I found epoxy to be very absorbtive and could be damaged during
> > 2-photon imaging.
> > >
> > > Regards,
> > > Glen MacDonald
> > > Digital Microscopy Center
> > > Box 357923
> > > University of Washington
> > > Seattle, WA 98195-7923  USA
> > > [phone]
> > > [hidden email]
> > >
> > >
> > >
> > >
> > >
> > >
> > >
> > >
> > >> On May 27, 2016, at 8:03 AM, Nuno Moreno
> > >> <[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.
> > >> *****
> > >>
> > >> Dear Craig
> > >>
> > >> I sent already a message directly to Joshua with a similar statement.
> > The problem is however when the burning in not at the first lens but
> > somewhere inside. In this case this means that the objective was not
> > assembled in a proper clean environment and has dust or other
> > particles inside. Actually here at the institute we were surprised
> > on how often
> this
> > happens from 2 different manufacturers. Maybe we were just unlucky.
> > If
> that
> > is the case I would ask for a replacement free of charge. I must
> > confess however that in Joshua case it does not look like but the
> > picture is not clear enough. I would advice you to bring your
> > objective to a
> stereoscope,
> > start to focus on the first back optical element and check whether
> > this
> is
> > superficial or not.
> > >>
> > >> Kind regards and good luck
> > >> Nuno Moreno
> > >>
> > >>> On 27 May 2016, at 15:07, Craig Brideau
> > >>> <[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.
> > >>> *****
> > >>>
> > >>> I had a glance at your picture. The black spots look like laser
> burns,
> > but
> > >>> there is also a clear bubble or edge that looks suspiciously
> > >>> like oil
> > also
> > >>> visible in the picture. It's possible you had some oil
> > >>> contamination
> > and
> > >>> then it burned onto the glass. Most optics will handle a
> > >>> surprising
> > amount
> > >>> of laser power as long as they are extremely clean. The instant
> > >>> there
> > is
> > >>> any dust/grease the laser will burn the surface as the
> > >>> contamination increases point absorption at the interface.
> > >>>
> > >>> Craig
> > >>>
> > >>>> On Fri, May 27, 2016 at 7:28 AM, Martin Wessendorf
> > >>>> <[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.
> > >>>> *****
> > >>>>
> > >>>> Dear Dr. Vaughan--
> > >>>>
> > >>>> Could it be dirt and oil on the lens, rather than being burned
> and/or
> > >>>> unglued?
> > >>>>
> > >>>> Martin Wessendorf
> > >>>>
> > >>>>
> > >>>>
> > >>>>
> > >>>>> On 5/27/2016 2:58 AM, Joshua Vaughan 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 damaged an objective lens, a Nikon CFI Plan Apo Lambda 100X
> > >>>>> Oil
> > 1.45 NA
> > >>>>> lens on our homebuilt TIRF/STORM microscope. Has anybody seen
> > >>>>> this
> > sort of
> > >>>>> thing before? I'm interested to hear, if so. We will repair or
> > replace it,
> > >>>>> and if replacing I wonder what to get. More details are below.
> > >>>>>
> > >>>>> Here is a link to a pic of part of the back focal plane. It
> > >>>>> has
> some
> > dark
> > >>>>> spots that look 'burned' and some stuff that looks like 'bubbles'.
> > >>>>>
> > >>>>> http://i63.tinypic.com/mmemvl.jpg
> > >>>>>
> > >>>>>
> > >>>>> Repairing this lens will cost >50% of a new lens so I want to
> decide
> > >>>>> whether
> > >>>>> to repair or to just get something else. I'd prefer something
> > >>>>> with
> a
> > >>>>> higher
> > >>>>> damage threshold but I was unsuccessful in getting data on
> > >>>>> lens
> > damage
> > >>>>> thresholds.
> > >>>>>
> > >>>>> If replacing, what to get? I noticed three lenses online and I
> > wonder what
> > >>>>> is best for STORM and whether all have a similar damage
> > >>>>> threshold
> or
> > not.
> > >>>>>
> > >>>>> 1. CFI Plan Apo Lambda 100X Oil 1.45 NA (current lens) 2. CFI
> > >>>>> Plan Apo Lambda DM 100X Oil 1.45 NA 3. CFI Apo SR TIRF 100X
> > >>>>> Oil 1.49 NA
> > >>>>>
> > >>>>>
> > >>>>> I think the Lambda lenses are supposed to be lower background,
> > though I've
> > >>>>> never tested whether that is important or not. They are also
> > >>>>> Plan
> > lenses,
> > >>>>> but I'm not sure how important that is for the <40 micron
> > >>>>> regions
> we
> > >>>>> typically image. The SR lens has a correction collar which
> > >>>>> might be
> > useful
> > >>>>> (?) when imaging a bit off the glass. The online information
> > >>>>> says
> > the SR
> > >>>>> lens has a PSF optimized for STORM which sounds good but I'm
> > >>>>> not
> > sure what
> > >>>>> it actually means. I don't have any damage threshold
> > >>>>> information
> > about
> > >>>>> these
> > >>>>> lenses. Oh, we are pretty sure the damage was caused when we
> > >>>>> sent
> > ~120 mW
> > >>>>> at
> > >>>>> 405 nm was sent to the lens in EPI and later in TIRF. We plan
> > >>>>> to substantially attenuate the laser in the future, which
> > >>>>> didn't need
> > to be
> > >>>>> that big anyway, but I am not sure how much to attenuate by.
> > >>>>>
> > >>>>>
> > >>>>> Joshua C. Vaughan
> > >>>>> Assistant Professor
> > >>>>> Department of Chemistry
> > >>>>> Box 351700
> > >>>>> University of Washington
> > >>>>> Seattle, WA 98195
> > >>>>
> > >>>> --
> > >>>> Martin Wessendorf, Ph.D.                   office: [phone]
> > >>>> Assoc Prof, Dept Neuroscience                 lab: [phone]
> > >>>> University of Minnesota             Preferred FAX: [phone]
> > >>>> 6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
> > >>>> Minneapolis, MN  55455                    e-mail: [hidden email]
> > >>>>
> >
>

--
Joshua C. Vaughan
Assistant Professor
Department of Chemistry
Box 351700
University of Washington
Seattle, WA 98195
206-543-4644

[Andreas Bruckbauer]

*****
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.
*****

Thanks for pointing this out, I must have looked at the transmission curve of an old Zeiss lens, so it might be worth  updating to a newer TIRF lens!

Best wishes

Andreas

-----Original Message-----
From: "Reece, Jeff (NIH/NIDDK) [E]" <[hidden email]>
Sent: ‎08/‎06/‎2016 03:35
To: "[hidden email]" <[hidden email]>
Subject: Re: objective lens laser damage

I thought Zeiss might say something, but I believe the latest 100x TIRF lens will have similar %T to the Olympus.
You can compare a few Zeiss TIRF lenses here:
https://www.micro-shop.zeiss.com/?s=306493725d39c2&l=en&p=us&f=o&a=c&m=s&pid[]=420780-9970-000&pid[]=420792-9800-000&pid[]=420792-9800-720&o=&st=1

Cheers,
Jeff

-----Original Message-----
From: Andreas Bruckbauer [mailto:[hidden email]]
Sent: Tuesday, May 31, 2016 3:56 AM
To: [hidden email]
Subject: Re: objective lens laser damage

*****
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 Joshua,


several years ago, i discussed the maximum laser power one could focus at the back focal plane of an objective with an optical engineer from Olympus. At that time they just had increased the laser power of their TIRF lasers from 100 mW to 200 mW for 488 nm and that was regarded as save for their TIRF objectives (this is usually the power before coupling into the optical fibre and you expect 70% of that . We discussed the case of the OMX microscope, which uses Olympus objectives and higher laser powers (around 300 - 500mW) and he was concerned about possible damage, mainly of the coating. However in this case I think the laser is not focused on the BF plane. The Bruker Vutara microscope uses Olympus lenses and powers of 1000 mW for 488, 561, 642 nm but "only" 100 mW at 405 nm. But as i understand this is usually set up as a WF microscope for 3D imaging, so they also do not need to focus to a point. It is also interesting to look at the objective transmission data when available. The Olympus 100x TIRF objective e.g. has > 80% transmission from 400 - 700nm, while the Zeiss 100x TIRF lens starts with only 50% at 400 nm and stays below 70% for the visible spectrum. Unfortunately i could not find the data for Nikon or Leica objectives.


best wishes


Andreas










-----Original Message-----
From: Joshua Vaughan <[hidden email]>
To: CONFOCALMICROSCOPY <[hidden email]>
Sent: Tue, 31 May 2016 1:30
Subject: Re: objective lens laser damage

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*****

Thanks for several thoughtful replies. I have a couple of follow up points.

Is anyone aware of damage threshold data for objective lenses, from Nikon or other manufacturers? It seems many people suspect glue damage for my lens. I asked my local Nikon sales rep but he was unable to get data for me.

Does anybody have a favorite objective lens for STORM/PALM among the following Nikon lenses? I'm intrigued by #4, below, which is rated for high power (Seamus mentioned).

1. CFI Plan Apo Lambda 100X Oil 1.45 NA (current lens) 2. CFI Plan Apo Lambda DM 100X Oil 1.45 NA 3. CFI Apo SR TIRF 100X Oil 1.49 NA 4. CFI HP Apochromat TIRF 100x oil 1.49 NA


To John Oreopoulos... No, we don't need 1W lasers for imaging. We generally work with up to 50-100 mW for the 488/561/647/750 nm lines and use the rest of the laser power for other stuff including spectroscopy experiments. For the 405 nm line, it is typically below 1 mW, but considering our test which showed 405 nm is likely the problem, I wonder if a user may have blasted with 405 nm at some point leading to damage that got worse with repeated use even at lower power.

On Fri, May 27, 2016 at 2:31 PM, S Ram <[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.
> *****
>
> The bubbles is something that I observed years ago (postdoc days) even
> with dry lenses (20x PApo; Zeiss) that has never been illuminated with a laser.
> At that time, we suspected that someone accidentally used oil on the
> dry lens and forgot to wipe it off and was never noticed for a very long time.
> It is possible that it could well be that something internal was damaged.
>
> Sripad
>
>
> On Fri, May 27, 2016 at 10:41 AM, John Oreopoulos <
> [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.
> > *****
> >
> > Do you really need these high laser powers for your application?
> >
> > John Oreopoulos
> >
> > > On May 27, 2016, at 11:42 AM, Glen MacDonald
> > > <[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.
> > > *****
> > >
> > > Possibly the damage is to the optical cement between the lens elements.
> > I found epoxy to be very absorbtive and could be damaged during
> > 2-photon imaging.
> > >
> > > Regards,
> > > Glen MacDonald
> > > Digital Microscopy Center
> > > Box 357923
> > > University of Washington
> > > Seattle, WA 98195-7923  USA
> > > [phone]
> > > [hidden email]
> > >
> > >
> > >
> > >
> > >
> > >
> > >
> > >
> > >> On May 27, 2016, at 8:03 AM, Nuno Moreno
> > >> <[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.
> > >> *****
> > >>
> > >> Dear Craig
> > >>
> > >> I sent already a message directly to Joshua with a similar statement.
> > The problem is however when the burning in not at the first lens but
> > somewhere inside. In this case this means that the objective was not
> > assembled in a proper clean environment and has dust or other
> > particles inside. Actually here at the institute we were surprised
> > on how often
> this
> > happens from 2 different manufacturers. Maybe we were just unlucky.
> > If
> that
> > is the case I would ask for a replacement free of charge. I must
> > confess however that in Joshua case it does not look like but the
> > picture is not clear enough. I would advice you to bring your
> > objective to a
> stereoscope,
> > start to focus on the first back optical element and check whether
> > this
> is
> > superficial or not.
> > >>
> > >> Kind regards and good luck
> > >> Nuno Moreno
> > >>
> > >>> On 27 May 2016, at 15:07, Craig Brideau
> > >>> <[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.
> > >>> *****
> > >>>
> > >>> I had a glance at your picture. The black spots look like laser
> burns,
> > but
> > >>> there is also a clear bubble or edge that looks suspiciously
> > >>> like oil
> > also
> > >>> visible in the picture. It's possible you had some oil
> > >>> contamination
> > and
> > >>> then it burned onto the glass. Most optics will handle a
> > >>> surprising
> > amount
> > >>> of laser power as long as they are extremely clean. The instant
> > >>> there
> > is
> > >>> any dust/grease the laser will burn the surface as the
> > >>> contamination increases point absorption at the interface.
> > >>>
> > >>> Craig
> > >>>
> > >>>> On Fri, May 27, 2016 at 7:28 AM, Martin Wessendorf
> > >>>> <[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.
> > >>>> *****
> > >>>>
> > >>>> Dear Dr. Vaughan--
> > >>>>
> > >>>> Could it be dirt and oil on the lens, rather than being burned
> and/or
> > >>>> unglued?
> > >>>>
> > >>>> Martin Wessendorf
> > >>>>
> > >>>>
> > >>>>
> > >>>>
> > >>>>> On 5/27/2016 2:58 AM, Joshua Vaughan 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 damaged an objective lens, a Nikon CFI Plan Apo Lambda 100X
> > >>>>> Oil
> > 1.45 NA
> > >>>>> lens on our homebuilt TIRF/STORM microscope. Has anybody seen
> > >>>>> this
> > sort of
> > >>>>> thing before? I'm interested to hear, if so. We will repair or
> > replace it,
> > >>>>> and if replacing I wonder what to get. More details are below.
> > >>>>>
> > >>>>> Here is a link to a pic of part of the back focal plane. It
> > >>>>> has
> some
> > dark
> > >>>>> spots that look 'burned' and some stuff that looks like 'bubbles'.
> > >>>>>
> > >>>>> http://i63.tinypic.com/mmemvl.jpg
> > >>>>>
> > >>>>>
> > >>>>> Repairing this lens will cost >50% of a new lens so I want to
> decide
> > >>>>> whether
> > >>>>> to repair or to just get something else. I'd prefer something
> > >>>>> with
> a
> > >>>>> higher
> > >>>>> damage threshold but I was unsuccessful in getting data on
> > >>>>> lens
> > damage
> > >>>>> thresholds.
> > >>>>>
> > >>>>> If replacing, what to get? I noticed three lenses online and I
> > wonder what
> > >>>>> is best for STORM and whether all have a similar damage
> > >>>>> threshold
> or
> > not.
> > >>>>>
> > >>>>> 1. CFI Plan Apo Lambda 100X Oil 1.45 NA (current lens) 2. CFI
> > >>>>> Plan Apo Lambda DM 100X Oil 1.45 NA 3. CFI Apo SR TIRF 100X
> > >>>>> Oil 1.49 NA
> > >>>>>
> > >>>>>
> > >>>>> I think the Lambda lenses are supposed to be lower background,
> > though I've
> > >>>>> never tested whether that is important or not. They are also
> > >>>>> Plan
> > lenses,
> > >>>>> but I'm not sure how important that is for the <40 micron
> > >>>>> regions
> we
> > >>>>> typically image. The SR lens has a correction collar which
> > >>>>> might be
> > useful
> > >>>>> (?) when imaging a bit off the glass. The online information
> > >>>>> says
> > the SR
> > >>>>> lens has a PSF optimized for STORM which sounds good but I'm
> > >>>>> not
> > sure what
> > >>>>> it actually means. I don't have any damage threshold
> > >>>>> information
> > about
> > >>>>> these
> > >>>>> lenses. Oh, we are pretty sure the damage was caused when we
> > >>>>> sent
> > ~120 mW
> > >>>>> at
> > >>>>> 405 nm was sent to the lens in EPI and later in TIRF. We plan
> > >>>>> to substantially attenuate the laser in the future, which
> > >>>>> didn't need
> > to be
> > >>>>> that big anyway, but I am not sure how much to attenuate by.
> > >>>>>
> > >>>>>
> > >>>>> Joshua C. Vaughan
> > >>>>> Assistant Professor
> > >>>>> Department of Chemistry
> > >>>>> Box 351700
> > >>>>> University of Washington
> > >>>>> Seattle, WA 98195
> > >>>>
> > >>>> --
> > >>>> Martin Wessendorf, Ph.D.                   office: [phone]
> > >>>> Assoc Prof, Dept Neuroscience                 lab: [phone]
> > >>>> University of Minnesota             Preferred FAX: [phone]
> > >>>> 6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
> > >>>> Minneapolis, MN  55455                    e-mail: [hidden email]
> > >>>>
> >
>

--
Joshua C. Vaughan
Assistant Professor
Department of Chemistry
Box 351700
University of Washington
Seattle, WA 98195
206-543-4644