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Automated liquid immersion

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Csúcs Gábor-3

Automated liquid immersion

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Dear All,

Following up the previous discussion about automated water/oil immersion, just some information. Besides the mentioned EMBL solution there is also a general (for all microscope brands) solution available from Marzhauser (https://www.marzhauser.com/en/products/liquid-immersion.html). I have no personal experience with it just has seen it at an exhibition. Looked quite nice.

Greetings Gabor

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Francesco Pasqualini
Sent: 08 January 2020 13:16
To: [hidden email]
Subject: Re: Spinning disk or (new) point scanning confocals for live imaging of 3D engineered tissues?

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Hi Sylvie - I am also very interested but I don't seem to be able to find the product at https://embl-em.de/ - do you have any tips on how to get the device? Thanks, Francesco

On Wed, Jan 8, 2020 at 12:57 PM VERMEREN Matthieu < [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 Sylvie,
>
> I was not aware of a dispenser for oil or silicone, thanks for the tip
> :-)
>
> How do you avoid spillage?
>
> Sincerely,
>
> Matthieu
>
> -----Original Message-----
> From: Confocal Microscopy List <[hidden email]> On
> Behalf Of Sylvie Le Guyader
> Sent: Sunday, 05 January, 2020 13:25
> To: [hidden email]
> Subject: Re: Spinning disk or (new) point scanning confocals for live
> imaging of 3D engineered tissues?
>
> *****
> 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 Mattieu
>
> I am curious to know why it would not be possible to use JOBS with the
> silicon objective. To my knowledge you can use JOBS with any objectives.
> Do you perhaps mean that you cannot image a multiwell plate with an
> oil objective? If this is the case, I can say that it is possible
> since we do it at our facility. You need an oil dispenser for your
> objective. We bought one quite a long time ago from EMBLEM, the
> commercial side of EMBL ( https://embl-em.de/). It works without problem.
>
> 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: +46 (0) 73 733 5008
> LCI website
> LCI microscopy blog
>
> -----Original Message-----
> From: Confocal Microscopy List <[hidden email]> On
> Behalf Of VERMEREN Matthieu
> Sent: Sunday, 5 January, 2020 13:55
> To: [hidden email]
> Subject: Re: Spinning disk or (new) point scanning confocals for live
> imaging of 3D engineered tissues?
>
> *****
> 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&data=02%7C01%7Cs
> ylvie.le.guyader%40KI.SE%7Caab63e63cdef4257be8808d791e1b4ff%7Cbff7eef1
> cf4b4f32be3da1dda043c05d%7C0%7C0%7C637138270916080122&sdata=QhP9C5
> P28OGIPkTfW2fQhHK%2FCIjgVN43KgQgF%2BHLgak%3D&reserved=0
> Post images on
> https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.i
> mgur.com&data=02%7C01%7Csylvie.le.guyader%40KI.SE%7Caab63e63cdef42
> 57be8808d791e1b4ff%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637138
> 270916080122&sdata=am3dgas9TvzghJ%2BaNk2zd8BpYAanylijfCPzI8z0MQ4%3
> D&reserved=0
> and include the link in your posting.
> *****
>
> Hi Francesco,
>
> We have a Nikon A1R on Ti2 on demo, resonant scanning combined with
> Nikon's new silicone lenses (long working distance, better match to
> live
> system) means we get excellent images on thick immunostained samples
> (organoids in matrix, tissue slices, young embryos). An organoid that
> would take ~30 minutes to image on a standard system is done in less
> than 6 minutes and with very little noise... meaning some experiments
> that were going to be too long and too expensive are now considered
> do-able by a fair few post docs and PIs. The advantage is that we can
> also use Nikon's JOB for multiwell system (sadly not with the silicone
> lenses, but water lenses are an option).
>
> I'm hoping to test it on Ca++ imaging or on FRET.
>
> Sincerely,
>
> Matthieu
>
> -----Original Message-----
> From: Confocal Microscopy List <[hidden email]> On
> Behalf Of Francesco Pasqualini
> Sent: 13 December 2019 17:18
> To: [hidden email]
> Subject: Spinning disk or (new) point scanning confocals for live
> imaging of 3D engineered tissues?
>
> *****
> 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&data=02%7C01%7Cs
> ylvie.le.guyader%40KI.SE%7Caab63e63cdef4257be8808d791e1b4ff%7Cbff7eef1
> cf4b4f32be3da1dda043c05d%7C0%7C0%7C637138270916080122&sdata=QhP9C5
> P28OGIPkTfW2fQhHK%2FCIjgVN43KgQgF%2BHLgak%3D&reserved=0
> Post images on
> https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.i
> mgur.com&data=02%7C01%7Csylvie.le.guyader%40KI.SE%7Caab63e63cdef42
> 57be8808d791e1b4ff%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637138
> 270916090122&sdata=9xFFxI0njDr%2B7nH3owxD%2FSlALKPchC09wMTk1PKGouM
> %3D&reserved=0
> and include the link in your posting.
> *****
>
> Hi,
> I will start my ERC-funded lab in Italy in 2020, which means I am
> fortunate to be shopping for a confocal instrument right about now.
>
> Since I do mostly live experiments (more details below) I was going to
> get a spinning disk system. But, I realized that the new scanning
> confocals are also relatively fast and gentle. The question is how
> much (if at all) slower and harsher are they with respect to spinning disks?
>
> More details:
> - I do a lot of live experiments (traction force microscopy and
> voltage/calcium-sensitive dyes) on engineered cells (300x300 um FOV)
> and tissues (600x600 um FOV) in 2D or 3D (<300 um thick) that I
> complement with immunostainings after fixation. Needed acquisition
> rates go from <1 fps to 100s depending on the application.
>
> - Originally, I was oriented towards a spinning disk confocal
> (Yokogawa/SORA, Crest) and was looking at ways to deal with the issues
> of confocality on the 3D tissue (600x600x300 um volumes) case. But, I
> realized all vendors have new point scanning confocal (980+Airyscan2,
> FV3000, A1R) with resonant scanners that acquire full frames/ROIs at
> 10s/100s of fps
>
> - Of course, structured illumination (Elyra-7, N-SIM) and light-sheet
> microscopes (QuVi) are also appealing but relatively untested in my
> applications of interests....
>
> The application specialists from all vendors in my area have been
> great to work with but since my lab is not up and running, yet, I
> can't demo these systems directly. Therefore, I could use help and
> feedback, especially from people who have had related experiences in the recent past.
>
> Thanks,
> Francesco
>
> --
> Francesco S. Pasqualini
> Visiting Professor University of Pavia Associate Harvard University
>
> tel: +39 351-521-7788 (IT)
> tel: +1 617-401-5243 (USA)
> The University of Edinburgh is a charitable body, registered in
> Scotland, with registration number SC005336.
>
>
> 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>.
>

--
Francesco S. Pasqualini
Visiting Professor University of Pavia
Associate Harvard University

tel: +39 351-521-7788 (IT)
tel: +1 617-401-5243 (USA)

Sylvie Le Guyader

Re: Automated liquid immersion

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http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
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*****

Very interesting Gabor! Thanks for sharing!
For the EMBLEM dispenser, we bought it about 9 years ago. I think the best is to contact them and ask if they still manufacture it. One had to send the objective to them for a while as they make the cap to measure. It looks very similar to the one the Gabor pointed to. The cap sits on top of the objective tightly. The very top fills with oil which is constantly added/removed. In our hands it doesn’t spill.
Here is where to contact EMBLEM: https://embl-em.de/

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: +46 (0) 73 733 5008
LCI website
Follow our microscopy blog!

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Csucs Gabor
Sent: 08 January 2020 13:26
To: [hidden email]
Subject: Automated liquid immersion

*****
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Post images on https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&data=02%7C01%7Csylvie.le.guyader%40KI.SE%7Cd4ab70fb9c02429764c408d7943822b8%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637140841235785069&sdata=qwgEX2e5jxyeizkTlvb65GlwdVwxpORGx4WDylanO%2F0%3D&reserved=0 and include the link in your posting.
*****

Dear All,

Following up the previous discussion about automated water/oil immersion, just some information. Besides the mentioned EMBL solution there is also a general (for all microscope brands) solution available from Marzhauser (https://eur01.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.marzhauser.com%2Fen%2Fproducts%2Fliquid-immersion.html&data=02%7C01%7Csylvie.le.guyader%40KI.SE%7Cd4ab70fb9c02429764c408d7943822b8%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637140841235795061&sdata=s0U7emYX10unDuOsvIN8s2bv93ZIam%2FYAkiRXUsX%2BLw%3D&reserved=0). I have no personal experience with it just has seen it at an exhibition. Looked quite nice.

Greetings Gabor

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Francesco Pasqualini
Sent: 08 January 2020 13:16
To: [hidden email]
Subject: Re: Spinning disk or (new) point scanning confocals for live imaging of 3D engineered tissues?

*****
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&data=02%7C01%7Csylvie.le.guyader%40KI.SE%7Cd4ab70fb9c02429764c408d7943822b8%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637140841235795061&sdata=eMV8UT5jAPrJZPbSpjAMDcdOq3sv2zjNQmDZ5epnpGk%3D&reserved=0
Post images on https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&data=02%7C01%7Csylvie.le.guyader%40KI.SE%7Cd4ab70fb9c02429764c408d7943822b8%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637140841235795061&sdata=LKfjnT0j82H9Q4owHN0hOQWyNRF9WzR8Kfeyxrjzhx4%3D&reserved=0 and include the link in your posting.
*****

Hi Sylvie - I am also very interested but I don't seem to be able to find the product at https://eur01.safelinks.protection.outlook.com/?url=https%3A%2F%2Fembl-em.de%2F&data=02%7C01%7Csylvie.le.guyader%40KI.SE%7Cd4ab70fb9c02429764c408d7943822b8%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637140841235795061&sdata=RoCuz1FNm65Ya0vvEtac20oMhngOe5fKJCilxJAr4ZU%3D&reserved=0 - do you have any tips on how to get the device? Thanks, Francesco

On Wed, Jan 8, 2020 at 12:57 PM VERMEREN Matthieu < [hidden email]> wrote:

> *****
> 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&data=02%7C01%7Cs
> ylvie.le.guyader%40KI.SE%7Cd4ab70fb9c02429764c408d7943822b8%7Cbff7eef1
> cf4b4f32be3da1dda043c05d%7C0%7C0%7C637140841235795061&sdata=eMV8UT
> 5jAPrJZPbSpjAMDcdOq3sv2zjNQmDZ5epnpGk%3D&reserved=0
> Post images on https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&data=02%7C01%7Csylvie.le.guyader%40KI.SE%7Cd4ab70fb9c02429764c408d7943822b8%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637140841235795061&sdata=LKfjnT0j82H9Q4owHN0hOQWyNRF9WzR8Kfeyxrjzhx4%3D&reserved=0 and include the link in your posting.
> *****
>
> Hi Sylvie,
>
> I was not aware of a dispenser for oil or silicone, thanks for the tip
> :-)
>
> How do you avoid spillage?
>
> Sincerely,
>
> Matthieu
>
> -----Original Message-----
> From: Confocal Microscopy List <[hidden email]> On
> Behalf Of Sylvie Le Guyader
> Sent: Sunday, 05 January, 2020 13:25
> To: [hidden email]
> Subject: Re: Spinning disk or (new) point scanning confocals for live
> imaging of 3D engineered tissues?
>
> *****
> 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&data=02%7C01%7Cs
> ylvie.le.guyader%40KI.SE%7Cd4ab70fb9c02429764c408d7943822b8%7Cbff7eef1
> cf4b4f32be3da1dda043c05d%7C0%7C0%7C637140841235795061&sdata=eMV8UT
> 5jAPrJZPbSpjAMDcdOq3sv2zjNQmDZ5epnpGk%3D&reserved=0
> Post images on https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&data=02%7C01%7Csylvie.le.guyader%40KI.SE%7Cd4ab70fb9c02429764c408d7943822b8%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637140841235795061&sdata=LKfjnT0j82H9Q4owHN0hOQWyNRF9WzR8Kfeyxrjzhx4%3D&reserved=0 and include the link in your posting.
> *****
>
> Hi Mattieu
>
> I am curious to know why it would not be possible to use JOBS with the
> silicon objective. To my knowledge you can use JOBS with any objectives.
> Do you perhaps mean that you cannot image a multiwell plate with an
> oil objective? If this is the case, I can say that it is possible
> since we do it at our facility. You need an oil dispenser for your
> objective. We bought one quite a long time ago from EMBLEM, the
> commercial side of EMBL ( https://eur01.safelinks.protection.outlook.com/?url=https%3A%2F%2Fembl-em.de%2F&data=02%7C01%7Csylvie.le.guyader%40KI.SE%7Cd4ab70fb9c02429764c408d7943822b8%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637140841235795061&sdata=RoCuz1FNm65Ya0vvEtac20oMhngOe5fKJCilxJAr4ZU%3D&reserved=0). It works without problem.
>
> 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: +46 (0) 73 733 5008
> LCI website
> LCI microscopy blog
>
> -----Original Message-----
> From: Confocal Microscopy List <[hidden email]> On
> Behalf Of VERMEREN Matthieu
> Sent: Sunday, 5 January, 2020 13:55
> To: [hidden email]
> Subject: Re: Spinning disk or (new) point scanning confocals for live
> imaging of 3D engineered tissues?
>
> *****
> 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&data=02%7C01%7Cs
> ylvie.le.guyader%40KI.SE%7Caab63e63cdef4257be8808d791e1b4ff%7Cbff7eef1
> cf4b4f32be3da1dda043c05d%7C0%7C0%7C637138270916080122&sdata=QhP9C5
> P28OGIPkTfW2fQhHK%2FCIjgVN43KgQgF%2BHLgak%3D&reserved=0
> Post images on
> https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.i
> mgur.com&data=02%7C01%7Csylvie.le.guyader%40KI.SE%7Caab63e63cdef42
> 57be8808d791e1b4ff%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637138
> 270916080122&sdata=am3dgas9TvzghJ%2BaNk2zd8BpYAanylijfCPzI8z0MQ4%3
> D&reserved=0
> and include the link in your posting.
> *****
>
> Hi Francesco,
>
> We have a Nikon A1R on Ti2 on demo, resonant scanning combined with
> Nikon's new silicone lenses (long working distance, better match to
> live
> system) means we get excellent images on thick immunostained samples
> (organoids in matrix, tissue slices, young embryos). An organoid that
> would take ~30 minutes to image on a standard system is done in less
> than 6 minutes and with very little noise... meaning some experiments
> that were going to be too long and too expensive are now considered
> do-able by a fair few post docs and PIs. The advantage is that we can
> also use Nikon's JOB for multiwell system (sadly not with the silicone
> lenses, but water lenses are an option).
>
> I'm hoping to test it on Ca++ imaging or on FRET.
>
> Sincerely,
>
> Matthieu
>
> -----Original Message-----
> From: Confocal Microscopy List <[hidden email]> On
> Behalf Of Francesco Pasqualini
> Sent: 13 December 2019 17:18
> To: [hidden email]
> Subject: Spinning disk or (new) point scanning confocals for live
> imaging of 3D engineered tissues?
>
> *****
> 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&data=02%7C01%7Cs
> ylvie.le.guyader%40KI.SE%7Caab63e63cdef4257be8808d791e1b4ff%7Cbff7eef1
> cf4b4f32be3da1dda043c05d%7C0%7C0%7C637138270916080122&sdata=QhP9C5
> P28OGIPkTfW2fQhHK%2FCIjgVN43KgQgF%2BHLgak%3D&reserved=0
> Post images on
> https://eur01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.i
> mgur.com&data=02%7C01%7Csylvie.le.guyader%40KI.SE%7Caab63e63cdef42
> 57be8808d791e1b4ff%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637138
> 270916090122&sdata=9xFFxI0njDr%2B7nH3owxD%2FSlALKPchC09wMTk1PKGouM
> %3D&reserved=0
> and include the link in your posting.
> *****
>
> Hi,
> I will start my ERC-funded lab in Italy in 2020, which means I am
> fortunate to be shopping for a confocal instrument right about now.
>
> Since I do mostly live experiments (more details below) I was going to
> get a spinning disk system. But, I realized that the new scanning
> confocals are also relatively fast and gentle. The question is how
> much (if at all) slower and harsher are they with respect to spinning disks?
>
> More details:
> - I do a lot of live experiments (traction force microscopy and
> voltage/calcium-sensitive dyes) on engineered cells (300x300 um FOV)
> and tissues (600x600 um FOV) in 2D or 3D (<300 um thick) that I
> complement with immunostainings after fixation. Needed acquisition
> rates go from <1 fps to 100s depending on the application.
>
> - Originally, I was oriented towards a spinning disk confocal
> (Yokogawa/SORA, Crest) and was looking at ways to deal with the issues
> of confocality on the 3D tissue (600x600x300 um volumes) case. But, I
> realized all vendors have new point scanning confocal (980+Airyscan2,
> FV3000, A1R) with resonant scanners that acquire full frames/ROIs at
> 10s/100s of fps
>
> - Of course, structured illumination (Elyra-7, N-SIM) and light-sheet
> microscopes (QuVi) are also appealing but relatively untested in my
> applications of interests....
>
> The application specialists from all vendors in my area have been
> great to work with but since my lab is not up and running, yet, I
> can't demo these systems directly. Therefore, I could use help and
> feedback, especially from people who have had related experiences in the recent past.
>
> Thanks,
> Francesco
>
> --
> Francesco S. Pasqualini
> Visiting Professor University of Pavia Associate Harvard University
>
> tel: +39 351-521-7788 (IT)
> tel: +1 617-401-5243 (USA)
> The University of Edinburgh is a charitable body, registered in
> Scotland, with registration number SC005336.
>
>
> 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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>
> 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>.
>

--
Francesco S. Pasqualini
Visiting Professor University of Pavia
Associate Harvard University

tel: +39 351-521-7788 (IT)
tel: +1 617-401-5243 (USA)