Dipping objectives - water or PBS?

[Adam Glaser]

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Hi everyone,
I was wondering if anyone had experience or knew if water dipping objectives (specifically the Zeiss W Plan-Apochromat) were designed for immersion in DI water or PBS. Or if they have been designed to perform well in both refractive indices?
Thanks!
Adam

[weber]

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I'm working with this lens on in-vivo. We use both and I don't see much difference. But as I also work with a temp controlled chamber so most of the time I use ultrasound gel.

[Sylvie Le Guyader]

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Hi Adam

Do you find a difference in refraction index between PBS and water? I think they are both 1.33.
If they have the same refraction index then the objective should perform identically with both.

Med vänlig hälsning / Best regards
Sylvie
@@@@@@@@@@@@@@@@@@@@@@@@
Sylvie Le Guyader, PhD
Live Cell Imaging Facility Manager
Karolinska Institutet- Bionut Dpt
Blickagången 16,
Room 7362 (lab)/7840 (office)
14157 Huddinge, Sweden
mobile: [phone]
LCI website
LCI microscopy blog

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Adam Glaser
Sent: Friday, 7 February, 2020 06:42
To: [hidden email]
Subject: Dipping objectives - water or PBS?

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*****

Hi everyone,
I was wondering if anyone had experience or knew if water dipping objectives (specifically the Zeiss W Plan-Apochromat) were designed for immersion in DI water or PBS. Or if they have been designed to perform well in both refractive indices?
Thanks!
Adam


När du skickar e-post till Karolinska Institutet (KI) innebär detta att KI kommer att behandla dina personuppgifter. Här finns information om hur KI behandlar personuppgifter<https://ki.se/medarbetare/integritetsskyddspolicy>.


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[Craig Brideau]

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The salt doesn't make that much of a difference. One detail is to remind
users to rinse the lens off with pure water after using PBS. I made a
'house call' to a lab that was having problems using one of their patch
rigs. After checking all their filters and prisms I happened to look down
at the objective and found a perfect window of dried salt sitting over the
face of the objective. One lengthy freshwater soak later and everything was
fine...

Craig

On Fri, Feb 7, 2020 at 2:01 AM Sylvie Le Guyader <[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.
> *****
>
> Hi Adam
>
> Do you find a difference in refraction index between PBS and water? I
> think they are both 1.33.
> If they have the same refraction index then the objective should perform
> identically with both.
>
> Med vänlig hälsning / Best regards
> Sylvie
> @@@@@@@@@@@@@@@@@@@@@@@@
> Sylvie Le Guyader, PhD
> Live Cell Imaging Facility Manager
> Karolinska Institutet- Bionut Dpt
> Blickagången 16,
> Room 7362 (lab)/7840 (office)
> 14157 Huddinge, Sweden
> mobile: [phone]
> LCI website
> LCI microscopy blog
>
> -----Original Message-----
> From: Confocal Microscopy List <[hidden email]> On
> Behalf Of Adam Glaser
> Sent: Friday, 7 February, 2020 06:42
> To: [hidden email]
> Subject: Dipping objectives - water or PBS?
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
>
> https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=02%7C01%7Csylvie.le.guyader%40KI.SE%7C7633ffe7aec949b2022208d7ab908db5%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637166510111401463&amp;sdata=Us8B9MW12KnO0ZkHigDnV7dRMhaBFioHskNRbV2z6yg%3D&amp;reserved=0
> Post images on
> https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&amp;data=02%7C01%7Csylvie.le.guyader%40KI.SE%7C7633ffe7aec949b2022208d7ab908db5%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637166510111401463&amp;sdata=ow3oJfNxg%2FZ19nC57cb0RBwfQXdTgDRmEN7bxbNDgpA%3D&amp;reserved=0
> and include the link in your posting.
> *****
>
> Hi everyone,
> I was wondering if anyone had experience or knew if water dipping
> objectives (specifically the Zeiss W Plan-Apochromat) were designed for
> immersion in DI water or PBS. Or if they have been designed to perform well
> in both refractive indices?
> Thanks!
> Adam
>
>
> När du skickar e-post till Karolinska Institutet (KI) innebär detta att KI
> kommer att behandla dina personuppgifter. Här finns information om hur KI
> behandlar personuppgifter<
> https://ki.se/medarbetare/integritetsskyddspolicy>.
>
>
> Sending email to Karolinska Institutet (KI) will result in KI processing
> your personal data. You can read more about KI’s processing of personal
> data here<https://ki.se/en/staff/data-protection-policy>.
>

[Adam Glaser]

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Thanks everyone. I also now see online that PBS should have a RI of 1.335.
If this is true then I agree it is a very small difference. Measuring some
PBS in our lab on a refractometer I got a reading of 1.341 which seemed
like it could start to have an impact. I will try to contact Zeiss and see
what they designed the objective for, otherwise I'll just go with DI water
for now!

On Fri, Feb 7, 2020 at 10:08 AM 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.
> *****
>
> The salt doesn't make that much of a difference. One detail is to remind
> users to rinse the lens off with pure water after using PBS. I made a
> 'house call' to a lab that was having problems using one of their patch
> rigs. After checking all their filters and prisms I happened to look down
> at the objective and found a perfect window of dried salt sitting over the
> face of the objective. One lengthy freshwater soak later and everything was
> fine...
>
> Craig
>
> On Fri, Feb 7, 2020 at 2:01 AM Sylvie Le Guyader <[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.
> > *****
> >
> > Hi Adam
> >
> > Do you find a difference in refraction index between PBS and water? I
> > think they are both 1.33.
> > If they have the same refraction index then the objective should perform
> > identically with both.
> >
> > Med vänlig hälsning / Best regards
> > Sylvie
> > @@@@@@@@@@@@@@@@@@@@@@@@
> > Sylvie Le Guyader, PhD
> > Live Cell Imaging Facility Manager
> > Karolinska Institutet- Bionut Dpt
> > Blickagången 16,
> > Room 7362 (lab)/7840 (office)
> > 14157 Huddinge, Sweden
> > mobile: [phone]
> > LCI website
> > LCI microscopy blog
> >
> > -----Original Message-----
> > From: Confocal Microscopy List <[hidden email]> On
> > Behalf Of Adam Glaser
> > Sent: Friday, 7 February, 2020 06:42
> > To: [hidden email]
> > Subject: Dipping objectives - water or PBS?
> >
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> >
> >
> https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=02%7C01%7Csylvie.le.guyader%40KI.SE%7C7633ffe7aec949b2022208d7ab908db5%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637166510111401463&amp;sdata=Us8B9MW12KnO0ZkHigDnV7dRMhaBFioHskNRbV2z6yg%3D&amp;reserved=0
> > Post images on
> >
> https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&amp;data=02%7C01%7Csylvie.le.guyader%40KI.SE%7C7633ffe7aec949b2022208d7ab908db5%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637166510111401463&amp;sdata=ow3oJfNxg%2FZ19nC57cb0RBwfQXdTgDRmEN7bxbNDgpA%3D&amp;reserved=0
> > and include the link in your posting.
> > *****
> >
> > Hi everyone,
> > I was wondering if anyone had experience or knew if water dipping
> > objectives (specifically the Zeiss W Plan-Apochromat) were designed for
> > immersion in DI water or PBS. Or if they have been designed to perform
> well
> > in both refractive indices?
> > Thanks!
> > Adam
> >
> >
> > När du skickar e-post till Karolinska Institutet (KI) innebär detta att
> KI
> > kommer att behandla dina personuppgifter. Här finns information om hur KI
> > behandlar personuppgifter<
> > https://ki.se/medarbetare/integritetsskyddspolicy>.
> >
> >
> > Sending email to Karolinska Institutet (KI) will result in KI processing
> > your personal data. You can read more about KI’s processing of personal
> > data here<https://ki.se/en/staff/data-protection-policy>.
> >
>

[Mark Sonders]

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A hint following on Craig's spur from the original PBS refractive index question....

Caked-on salt films on water immersion objectives that have formed from repeated exposures to PBS buffers appear be removed by dipping them into 0.1 N acetic acid solution for ~10 min and rinsing with no obvious deleterious effect.

-=-Mark

Department of Psychiatry
Columbia University
Division of Molecular Therapeutics
New York State Psychiatric Institute
New York, NY  10032

[Craig Brideau]

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Thanks Mark! I'll give that a try when I next encounter salt films. It
would probably be quicker than just a DI water soak.

Craig

On Sat, Feb 8, 2020 at 7:43 AM Mark Sonders <[hidden email]> wrote:

> A hint following on Craig's spur from the original PBS refractive index
> question....
>
> Caked-on salt films on water immersion objectives that have formed from
> repeated exposures to PBS buffers appear be removed by dipping them into
> 0.1 N acetic acid solution for ~10 min and rinsing with no obvious
> deleterious effect.
>
> -=-Mark
>
> Department of Psychiatry
> Columbia University
> Division of Molecular Therapeutics
> New York State Psychiatric Institute
> New York, NY  10032
>
>
>

[Zdenek Svindrych-2]

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Hi Adam, btw, what's your RI reading for DI water?
Just FYI, our routine PBS (137 mM NaCl, 2.7 mM KCl, 12 mM phosphate buffer
pH 7.4) has a RI of 1.3355, and pure water gives me 1.3335 on my
refractometer.
zdenek

On Sat, Feb 8, 2020 at 12:50 AM Adam Glaser <[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.
> *****
>
> Thanks everyone. I also now see online that PBS should have a RI of 1.335.
> If this is true then I agree it is a very small difference. Measuring some
> PBS in our lab on a refractometer I got a reading of 1.341 which seemed
> like it could start to have an impact. I will try to contact Zeiss and see
> what they designed the objective for, otherwise I'll just go with DI water
> for now!
>
> On Fri, Feb 7, 2020 at 10:08 AM 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.
> > *****
> >
> > The salt doesn't make that much of a difference. One detail is to remind
> > users to rinse the lens off with pure water after using PBS. I made a
> > 'house call' to a lab that was having problems using one of their patch
> > rigs. After checking all their filters and prisms I happened to look down
> > at the objective and found a perfect window of dried salt sitting over
> the
> > face of the objective. One lengthy freshwater soak later and everything
> was
> > fine...
> >
> > Craig
> >
> > On Fri, Feb 7, 2020 at 2:01 AM Sylvie Le Guyader <
> [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.
> > > *****
> > >
> > > Hi Adam
> > >
> > > Do you find a difference in refraction index between PBS and water? I
> > > think they are both 1.33.
> > > If they have the same refraction index then the objective should
> perform
> > > identically with both.
> > >
> > > Med vänlig hälsning / Best regards
> > > Sylvie
> > > @@@@@@@@@@@@@@@@@@@@@@@@
> > > Sylvie Le Guyader, PhD
> > > Live Cell Imaging Facility Manager
> > > Karolinska Institutet- Bionut Dpt
> > > Blickagången 16,
> > > Room 7362 (lab)/7840 (office)
> > > 14157 Huddinge, Sweden
> > > mobile: [phone]
> > > LCI website
> > > LCI microscopy blog
> > >
> > > -----Original Message-----
> > > From: Confocal Microscopy List <[hidden email]> On
> > > Behalf Of Adam Glaser
> > > Sent: Friday, 7 February, 2020 06:42
> > > To: [hidden email]
> > > Subject: Dipping objectives - water or PBS?
> > >
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > >
> > >
> >
> https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=02%7C01%7Csylvie.le.guyader%40KI.SE%7C7633ffe7aec949b2022208d7ab908db5%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637166510111401463&amp;sdata=Us8B9MW12KnO0ZkHigDnV7dRMhaBFioHskNRbV2z6yg%3D&amp;reserved=0
> > > Post images on
> > >
> >
> https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&amp;data=02%7C01%7Csylvie.le.guyader%40KI.SE%7C7633ffe7aec949b2022208d7ab908db5%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637166510111401463&amp;sdata=ow3oJfNxg%2FZ19nC57cb0RBwfQXdTgDRmEN7bxbNDgpA%3D&amp;reserved=0
> > > and include the link in your posting.
> > > *****
> > >
> > > Hi everyone,
> > > I was wondering if anyone had experience or knew if water dipping
> > > objectives (specifically the Zeiss W Plan-Apochromat) were designed for
> > > immersion in DI water or PBS. Or if they have been designed to perform
> > well
> > > in both refractive indices?
> > > Thanks!
> > > Adam
> > >
> > >
> > > När du skickar e-post till Karolinska Institutet (KI) innebär detta att
> > KI
> > > kommer att behandla dina personuppgifter. Här finns information om hur
> KI
> > > behandlar personuppgifter<
> > > https://ki.se/medarbetare/integritetsskyddspolicy>.
> > >
> > >
> > > Sending email to Karolinska Institutet (KI) will result in KI
> processing
> > > your personal data. You can read more about KI’s processing of personal
> > > data here<https://ki.se/en/staff/data-protection-policy>.
> > >
> >
>

--
--
Zdenek Svindrych, Ph.D.
Research Associate - Imaging Specialist
Department of Biochemistry and Cell Biology
Geisel School of Medicine at Dartmouth

[mmodel]

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The error of measurement is usually greater than 0.001, and it depends on the wavelength

http://www.philiplaven.com/p20.html
Refractive index<http://www.philiplaven.com/p20.html>
Refractive index of water. Most people would assume that the refractive index of water is known to a high degree of accuracy. However, as shown in Fig. 1, the published literature reveals significant differences in the values of refractive index of water for a given wavelength.
www.philiplaven.com


________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Zdenek Svindrych <[hidden email]>
Sent: Monday, February 10, 2020 9:34 AM
To: [hidden email] <[hidden email]>
Subject: Re: Dipping objectives - water or PBS?

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*****

Hi Adam, btw, what's your RI reading for DI water?
Just FYI, our routine PBS (137 mM NaCl, 2.7 mM KCl, 12 mM phosphate buffer
pH 7.4) has a RI of 1.3355, and pure water gives me 1.3335 on my
refractometer.
zdenek

On Sat, Feb 8, 2020 at 12:50 AM Adam Glaser <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> https://nam03.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=02%7C01%7Cmmodel%40KENT.EDU%7C86daf42d16d04892344508d7ae36e26f%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637169423064861033&amp;sdata=5KA7aQsukPtd0ZYhtBR4ggTkNgkaDA6fKva740XgqCE%3D&amp;reserved=0
> Post images on https://nam03.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&amp;data=02%7C01%7Cmmodel%40KENT.EDU%7C86daf42d16d04892344508d7ae36e26f%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637169423064861033&amp;sdata=cTom0QQw3sOAuwvINDDv6pAwuPhPenNnux7NVqpAPFY%3D&amp;reserved=0 and include the link in your posting.
> *****
>
> Thanks everyone. I also now see online that PBS should have a RI of 1.335.
> If this is true then I agree it is a very small difference. Measuring some
> PBS in our lab on a refractometer I got a reading of 1.341 which seemed
> like it could start to have an impact. I will try to contact Zeiss and see
> what they designed the objective for, otherwise I'll just go with DI water
> for now!
>
> On Fri, Feb 7, 2020 at 10:08 AM Craig Brideau <[hidden email]>
> wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > https://nam03.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=02%7C01%7Cmmodel%40KENT.EDU%7C86daf42d16d04892344508d7ae36e26f%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637169423064861033&amp;sdata=5KA7aQsukPtd0ZYhtBR4ggTkNgkaDA6fKva740XgqCE%3D&amp;reserved=0
> > Post images on https://nam03.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&amp;data=02%7C01%7Cmmodel%40KENT.EDU%7C86daf42d16d04892344508d7ae36e26f%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637169423064861033&amp;sdata=cTom0QQw3sOAuwvINDDv6pAwuPhPenNnux7NVqpAPFY%3D&amp;reserved=0 and include the link in your
> posting.
> > *****
> >
> > The salt doesn't make that much of a difference. One detail is to remind
> > users to rinse the lens off with pure water after using PBS. I made a
> > 'house call' to a lab that was having problems using one of their patch
> > rigs. After checking all their filters and prisms I happened to look down
> > at the objective and found a perfect window of dried salt sitting over
> the
> > face of the objective. One lengthy freshwater soak later and everything
> was
> > fine...
> >
> > Craig
> >
> > On Fri, Feb 7, 2020 at 2:01 AM Sylvie Le Guyader <
> [hidden email]>
> > wrote:
> >
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > > https://nam03.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=02%7C01%7Cmmodel%40KENT.EDU%7C86daf42d16d04892344508d7ae36e26f%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637169423064861033&amp;sdata=5KA7aQsukPtd0ZYhtBR4ggTkNgkaDA6fKva740XgqCE%3D&amp;reserved=0
> > > Post images on https://nam03.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&amp;data=02%7C01%7Cmmodel%40KENT.EDU%7C86daf42d16d04892344508d7ae36e26f%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637169423064861033&amp;sdata=cTom0QQw3sOAuwvINDDv6pAwuPhPenNnux7NVqpAPFY%3D&amp;reserved=0 and include the link in your
> > posting.
> > > *****
> > >
> > > Hi Adam
> > >
> > > Do you find a difference in refraction index between PBS and water? I
> > > think they are both 1.33.
> > > If they have the same refraction index then the objective should
> perform
> > > identically with both.
> > >
> > > Med vänlig hälsning / Best regards
> > > Sylvie
> > > @@@@@@@@@@@@@@@@@@@@@@@@
> > > Sylvie Le Guyader, PhD
> > > Live Cell Imaging Facility Manager
> > > Karolinska Institutet- Bionut Dpt
> > > Blickagången 16,
> > > Room 7362 (lab)/7840 (office)
> > > 14157 Huddinge, Sweden
> > > mobile: [phone]
> > > LCI website
> > > LCI microscopy blog
> > >
> > > -----Original Message-----
> > > From: Confocal Microscopy List <[hidden email]> On
> > > Behalf Of Adam Glaser
> > > Sent: Friday, 7 February, 2020 06:42
> > > To: [hidden email]
> > > Subject: Dipping objectives - water or PBS?
> > >
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > >
> > >
> >
> https://nam03.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=02%7C01%7Cmmodel%40KENT.EDU%7C86daf42d16d04892344508d7ae36e26f%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637169423064861033&amp;sdata=5KA7aQsukPtd0ZYhtBR4ggTkNgkaDA6fKva740XgqCE%3D&amp;reserved=0
> > > Post images on
> > >
> >
> https://nam03.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&amp;data=02%7C01%7Cmmodel%40KENT.EDU%7C86daf42d16d04892344508d7ae36e26f%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637169423064861033&amp;sdata=cTom0QQw3sOAuwvINDDv6pAwuPhPenNnux7NVqpAPFY%3D&amp;reserved=0
> > > and include the link in your posting.
> > > *****
> > >
> > > Hi everyone,
> > > I was wondering if anyone had experience or knew if water dipping
> > > objectives (specifically the Zeiss W Plan-Apochromat) were designed for
> > > immersion in DI water or PBS. Or if they have been designed to perform
> > well
> > > in both refractive indices?
> > > Thanks!
> > > Adam
> > >
> > >
> > > När du skickar e-post till Karolinska Institutet (KI) innebär detta att
> > KI
> > > kommer att behandla dina personuppgifter. Här finns information om hur
> KI
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> > >
> > >
> > > Sending email to Karolinska Institutet (KI) will result in KI
> processing
> > > your personal data. You can read more about KI’s processing of personal
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> > >
> >
>

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Zdenek Svindrych, Ph.D.
Research Associate - Imaging Specialist
Department of Biochemistry and Cell Biology
Geisel School of Medicine at Dartmouth