Super-resolution microscopy - feedback wanted from staff at core facilities

[Jacqueline Ross]

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Hi everyone,

I'm enjoying the current conversation on super-resolution, especially because we are currently investigating options for purchasing a super-resolution microscope for our core facility. As you are all aware, there are a number of different technologies, which all have pros/cons depending on the specimen/labelling. We are currently considering STED, SML (STORM/PALM), SIM, Airyscan, etc.

I am looking for feedback from people working in core facilities like ours where a large variety of research interests are supported, e.g. cultured cells/tissue sections, plant/animal/bacteria, etc. Generally, our approach is to train users to work independently rather than carrying out work for people. I have already had a few conversations with colleagues in Australia which have been very useful.

If you work in a core facility and have one or more super-resolution microscopes, I would love to hear from you. In particular, I would like to know how often the super-resolution microscopes are used in comparison to confocal for example. It would also be helpful to know whether researchers expectations of the technology are being met and what level of support is needed both from facility staff and from service engineers. Advice on data processing requirements and/or problems would also be of interest. We are consulting with our researchers via a survey and our feeling is that many of our respondents are not realistic about what might be required in order to get useful data. Of course everyone wants the highest possible resolution, z stack capability, >2 fluorophores and live cell imaging.

If you have more than one super-resolution system, which is more popular and why? This purchase will require significant investment by our institution and will require a business case so we want to make sure that we make the best decision for our facility.

Off list replies are welcomed and will be kept confidential to the members of our working group if necessary. If you would prefer to provide your views via telephone/skype, I'm happy to speak to you at a time that suits you.

Kind regards,

Jacqui


Jacqueline Ross
Biomedical Imaging Microscopist
Biomedical Imaging Research Unit (BIRU)
School of Medical Sciences
Faculty of Medical & Health Sciences
The University of Auckland
Private Bag 92019
Auckland 1142, NEW ZEALAND

Telephone: Ext 87438; DDI:  [phone]

Website:    http://www.auckland.ac.nz/biru

[Ferhan A]

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Dear List Members,

We are in the same boat with Jacqui (although Hungary is far from sea, we
have Lake Balaton)

In the near future, we also plan to invest in a super-resolution microscope
as our next big purchase and it will be great to hear comments regarding
this topic.

Best Regards,


Ferhan



Ferhan Ayaydin, Ph.D.

Cellular Imaging Laboratory

Biological Research Centre

Hungarian Academy of Sciences,

Szeged, Hungary

On Thu, Aug 24, 2017 at 4:04 AM, Jacqui Ross <[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 everyone,
>
> I'm enjoying the current conversation on super-resolution, especially
> because we are currently investigating options for purchasing a
> super-resolution microscope for our core facility. As you are all aware,
> there are a number of different technologies, which all have pros/cons
> depending on the specimen/labelling. We are currently considering STED, SML
> (STORM/PALM), SIM, Airyscan, etc.
>
> I am looking for feedback from people working in core facilities like ours
> where a large variety of research interests are supported, e.g. cultured
> cells/tissue sections, plant/animal/bacteria, etc. Generally, our approach
> is to train users to work independently rather than carrying out work for
> people. I have already had a few conversations with colleagues in Australia
> which have been very useful.
>
> If you work in a core facility and have one or more super-resolution
> microscopes, I would love to hear from you. In particular, I would like to
> know how often the super-resolution microscopes are used in comparison to
> confocal for example. It would also be helpful to know whether researchers
> expectations of the technology are being met and what level of support is
> needed both from facility staff and from service engineers. Advice on data
> processing requirements and/or problems would also be of interest. We are
> consulting with our researchers via a survey and our feeling is that many
> of our respondents are not realistic about what might be required in order
> to get useful data. Of course everyone wants the highest possible
> resolution, z stack capability, >2 fluorophores and live cell imaging.
>
> If you have more than one super-resolution system, which is more popular
> and why? This purchase will require significant investment by our
> institution and will require a business case so we want to make sure that
> we make the best decision for our facility.
>
> Off list replies are welcomed and will be kept confidential to the members
> of our working group if necessary. If you would prefer to provide your
> views via telephone/skype, I'm happy to speak to you at a time that suits
> you.
>
> Kind regards,
>
> Jacqui
>
>
> Jacqueline Ross
> Biomedical Imaging Microscopist
> Biomedical Imaging Research Unit (BIRU)
> School of Medical Sciences
> Faculty of Medical & Health Sciences
> The University of Auckland
> Private Bag 92019
> Auckland 1142, NEW ZEALAND
>
> Telephone: Ext 87438; DDI:  [phone]
>
> Website:    http://www.auckland.ac.nz/biru
>

[Cvic Innocent]

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*****

Hello Jacqui & Ferhan (& all)

For SIM the OMX series*** (notably the V3 & the V4, products of Applied
Precision bought by GE) is wonderfully reliable and, once your user base
has been trained, very very straight forward. I do warn you that it has a
large footprint... it's big. Recently there is the OMX-SR, with a smaller
footprint, the size of an ice machine, however I personally have not used
it. The V3 and V4 are also able to do TIRF if you have a TIRF module. I am
a huge fan of the instrument since it is easy for users to understand how
to use & to quickly use independently. Moreover, once you look "under the
hood" & become familiar with the components, core facility staff can
personally do some basic repairs on the instrument (I'm not saying void
your service contract... I'm just saying what is going on underneath is not
only obvious but accessible... makes customization fun!).

The Airyscan is also great but, when it has an issue (such as uneven
illumination) it becomes a black box. I have had equal number of colleagues
praise and "very much not praise" the instrument because a service
technician needed to live on site.

As an aside (& you had alluded to this but I will mention it again), core
facilities must insist that sample preparation is done well & *with the
super-res technique kept in mind*. Images can be pretty but must firstly be
quantifiable, and to get there is a holistic process where researchers
appreciate that the best quality images come from the best possible
starting conditions -- a process where sample preparation is absolutely
crucial.

For lightsheet I recommend the Z1 from Zeiss. Also short learning curve for
researchers... but be sure you have plenty of supplies (tubes, filaments,
chamber windows and DATA STORAGE!!!) to spare because once your base gets
comfortable the instrument can be used quite heavily.

For single-molecule tracking/localization I am a fan of the Nanoimager by
ONI (Oxford NanoInstruments). Compact, specialized and straight forward.

Usage
I would often sit with a user to determine the modality needed... because
by far many researchers would do well with confocal resolution. However, a
good number of research groups genuinely needed the precision afforded by
nanoscopy and I can tell you, these instruments were used just as much as
the confocals. But the term 'super-resolution' excites people so comb
through the crowd by assessing which types of projects really will benefit
from SR.

Lastly, if possible... make your own instrument. Microscopy development is
not daunting & is well worth the effort. Not everyone has space or manpower
or money for this but if you have even a little bit to spare, please
consider this :]

Let me know if you have more questions or if you wish to chat more offlist.

Cordially,
cvic


***be sure to negotiate a good service/repair contract if possible. GE may
be phasing that out for newer products



Cassandravictoria Innocent, PhD
Assistant Director of Microscopy
Cellular Imaging Core
IDDRC, BCH & HMS

---------- Forwarded message ----------
From: Ferhan A <[hidden email]>
Date: Thu, Aug 24, 2017 at 5:43 AM
Subject: Re: Super-resolution microscopy - feedback wanted from staff at
core facilities
To: [hidden email]


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Dear List Members,

We are in the same boat with Jacqui (although Hungary is far from sea, we
have Lake Balaton)

In the near future, we also plan to invest in a super-resolution microscope
as our next big purchase and it will be great to hear comments regarding
this topic.

Best Regards,


Ferhan



Ferhan Ayaydin, Ph.D.

Cellular Imaging Laboratory

Biological Research Centre

Hungarian Academy of Sciences,

Szeged, Hungary

On Thu, Aug 24, 2017 at 4:04 AM, Jacqui Ross <[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 everyone,
>
> I'm enjoying the current conversation on super-resolution, especially
> because we are currently investigating options for purchasing a
> super-resolution microscope for our core facility. As you are all aware,
> there are a number of different technologies, which all have pros/cons
> depending on the specimen/labelling. We are currently considering STED,

SML
> (STORM/PALM), SIM, Airyscan, etc.
>
> I am looking for feedback from people working in core facilities like ours
> where a large variety of research interests are supported, e.g. cultured
> cells/tissue sections, plant/animal/bacteria, etc. Generally, our approach
> is to train users to work independently rather than carrying out work for
> people. I have already had a few conversations with colleagues in
Australia

> which have been very useful.
>
> If you work in a core facility and have one or more super-resolution
> microscopes, I would love to hear from you. In particular, I would like to
> know how often the super-resolution microscopes are used in comparison to
> confocal for example. It would also be helpful to know whether researchers
> expectations of the technology are being met and what level of support is
> needed both from facility staff and from service engineers. Advice on data
> processing requirements and/or problems would also be of interest. We are
> consulting with our researchers via a survey and our feeling is that many
> of our respondents are not realistic about what might be required in order
> to get useful data. Of course everyone wants the highest possible
> resolution, z stack capability, >2 fluorophores and live cell imaging.
>
> If you have more than one super-resolution system, which is more popular
> and why? This purchase will require significant investment by our
> institution and will require a business case so we want to make sure that
> we make the best decision for our facility.
>
> Off list replies are welcomed and will be kept confidential to the members
> of our working group if necessary. If you would prefer to provide your
> views via telephone/skype, I'm happy to speak to you at a time that suits
> you.
>
> Kind regards,
>
> Jacqui
>
>
> Jacqueline Ross
> Biomedical Imaging Microscopist
> Biomedical Imaging Research Unit (BIRU)
> School of Medical Sciences
> Faculty of Medical & Health Sciences
> The University of Auckland
> Private Bag 92019
> Auckland 1142, NEW ZEALAND
>
> Telephone: Ext 87438; DDI:  [phone]
>
> Website:    http://www.auckland.ac.nz/biru
>

[Janina Hanne]

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----------------------- Commercial Response -------------------------

Dear Ferhan,

among the super-resolution techniques available today I believe STED microscopy is a particularly powerful one. I used it a lot during my PhD and was quite happy with it. It provides extremely high spatial resolution (40 nm and better) and, unlike many other methods, does not require any data postprocessing to give you an image.

If you are interested to get a personal impression of what STED can do (and how easy it is to operate a modern instrument) you are more than welcome to join the demo of our super-compact STEDYCON microscope at the University of Szeged in September. Please let me know if you are interested so we can make an appointment for a demo slot.
 
All the best,
Janina


---
Dr. Janina Hanne
Abberior Instruments GmbH
Phone: [phone]
email: [hidden email]
www.abberior-instruments.com

[Kyle Michael Douglass]

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*****

Hi Cvic,


On 08/24/2017 04:52 PM, Cvic Innocent wrote:
>
> Lastly, if possible... make your own instrument. Microscopy development is
> not daunting & is well worth the effort. Not everyone has space or manpower
> or money for this but if you have even a little bit to spare, please
> consider this :]
>

I like your summary a lot and would like to add one small tangential
comment about microscope homebuilding. If a research lab or a core
facility chooses to build their own setup, then I would highly advise
not to forget to factor in the effort required to develop and maintain
the control software. This part of microscope homebuilding is often
overlooked when planning for a new microscope and, in my experience,
more time consuming than working with the optics. Software difficulties
become especially acute in super-resolution, which often (but not
always) requires highly customized acquisition protocols.

Micro-Manager (https://www.micro-manager.org/) is a great open source
solution that helps alleviate some of these difficulties, but it likely
will still not be completely plug-and-play for many advanced microscope
modalities.

Cheers,
Kyle

--
Kyle M. Douglass, PhD
Post-doctoral researcher
The Laboratory of Experimental Biophysics
EPFL, Lausanne, Switzerland
http://kmdouglass.github.io
http://leb.epfl.ch

[Cvic Innocent]

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*****

I must agree with Kyle's addition: instrument control is half (...more like
60-70%) of the battle :]

Micro-manager is a great option. If the core wishes to be a bit more hands
off then there is always prepackaged LabView too. Additionally, using
Simulink integrated with Matlab is an option for a more hands-on approach &
of course people are welcome to create their own software too!

But for first-time instrument developers Micro-manager is a great option to
keep the entire affair short & simple.

Cordially,
cvic



On Thu, Aug 24, 2017 at 11:33 AM, Kyle Douglass <[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 Cvic,
>
>
> On 08/24/2017 04:52 PM, Cvic Innocent wrote:
>
>>
>> Lastly, if possible... make your own instrument. Microscopy development is
>> not daunting & is well worth the effort. Not everyone has space or
>> manpower
>> or money for this but if you have even a little bit to spare, please
>> consider this :]
>>
>>
> I like your summary a lot and would like to add one small tangential
> comment about microscope homebuilding. If a research lab or a core facility
> chooses to build their own setup, then I would highly advise not to forget
> to factor in the effort required to develop and maintain the control
> software. This part of microscope homebuilding is often overlooked when
> planning for a new microscope and, in my experience, more time consuming
> than working with the optics. Software difficulties become especially acute
> in super-resolution, which often (but not always) requires highly
> customized acquisition protocols.
>
> Micro-Manager (https://www.micro-manager.org/) is a great open source
> solution that helps alleviate some of these difficulties, but it likely
> will still not be completely plug-and-play for many advanced microscope
> modalities.
>
> Cheers,
> Kyle
>
> --
> Kyle M. Douglass, PhD
> Post-doctoral researcher
> The Laboratory of Experimental Biophysics
> EPFL, Lausanne, Switzerland
> http://kmdouglass.github.io
> http://leb.epfl.ch
>

[Ralph Bauer]

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***** Commercial Response ******

Hi Jacqui,

Along with the other instruments mentioned I would also suggest that you also consider the VisiTech, iSIM, which is based on Andrew York and Hari Shroff's work at NIH.

This is a real time, live cell, multi point confocal based on optical pixel reassignment. The resolution is similar to what you would see with "traditional" SIM instruments such as the OMX but only requires capturing a single image and does not require you to computationally construct your image from u multiple images.

When considering traditional SIM instruments I would recommend reading 'Strategic and practical guidelines for successful structured illumination microscopy" by Justin Demmerle et al.

I would also consider the mix of instruments a core facility has. Many cores have only laser scanning confocals but I think that there is an argument to be made for having more diversity of instrumentation available (such as a multi point confocal) if space and budget are available.

You also need to consider the time and resources the core has to train users and maintain the instruments they have. While I would love to have a Ferrari in my garage I can't afford to have Mario the mechanic live with me to keep it running.

This may make as system like the ONI Nanoimager which doesn't require alignment in the field a nice option if you require PALM/STORM capability.

Ralph Bauer
503-267-6818
www.BioVis.com

[Ferhan A]

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Dear list members,
Thank you very much for your on-list and off-list comments, help and
suggestions.They were very useful and will help us a lot during decision
making.
I also thank Jacqui for starting this discussion.
Have a nice weekend!
Ferhan

Ferhan Ayaydin, Ph.D.

Cellular Imaging Laboratory

Biological Research Centre

Hungarian Academy of Sciences,

Szeged, Hungary

On Thu, Aug 24, 2017 at 4:04 AM, Jacqui Ross <[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 everyone,
>
> I'm enjoying the current conversation on super-resolution, especially
> because we are currently investigating options for purchasing a
> super-resolution microscope for our core facility. As you are all aware,
> there are a number of different technologies, which all have pros/cons
> depending on the specimen/labelling. We are currently considering STED, SML
> (STORM/PALM), SIM, Airyscan, etc.
>
> I am looking for feedback from people working in core facilities like ours
> where a large variety of research interests are supported, e.g. cultured
> cells/tissue sections, plant/animal/bacteria, etc. Generally, our approach
> is to train users to work independently rather than carrying out work for
> people. I have already had a few conversations with colleagues in Australia
> which have been very useful.
>
> If you work in a core facility and have one or more super-resolution
> microscopes, I would love to hear from you. In particular, I would like to
> know how often the super-resolution microscopes are used in comparison to
> confocal for example. It would also be helpful to know whether researchers
> expectations of the technology are being met and what level of support is
> needed both from facility staff and from service engineers. Advice on data
> processing requirements and/or problems would also be of interest. We are
> consulting with our researchers via a survey and our feeling is that many
> of our respondents are not realistic about what might be required in order
> to get useful data. Of course everyone wants the highest possible
> resolution, z stack capability, >2 fluorophores and live cell imaging.
>
> If you have more than one super-resolution system, which is more popular
> and why? This purchase will require significant investment by our
> institution and will require a business case so we want to make sure that
> we make the best decision for our facility.
>
> Off list replies are welcomed and will be kept confidential to the members
> of our working group if necessary. If you would prefer to provide your
> views via telephone/skype, I'm happy to speak to you at a time that suits
> you.
>
> Kind regards,
>
> Jacqui
>
>
> Jacqueline Ross
> Biomedical Imaging Microscopist
> Biomedical Imaging Research Unit (BIRU)
> School of Medical Sciences
> Faculty of Medical & Health Sciences
> The University of Auckland
> Private Bag 92019
> Auckland 1142, NEW ZEALAND
>
> Telephone: Ext 87438; DDI:  [phone]
>
> Website:    http://www.auckland.ac.nz/biru
>

[Chloe van Oostende]

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Hi,
Being a plant biologist who is working with numerous sample during my
microscope facility experience (worms, yeast, human cells, tissue slices),
I can state with a strong confidence, that a super-resolution (SR)
microscope is under used.
We currently have a STORM, SIM, and STED. All under used because of
different reasons: time consuming to have the perfect slide to be imaged,
optimisation, time to get familiar with a software. Slides for STORM has to
have a low staining, STED requires high staining, SIM works well on a
restricted z-depth (multilayer of cells won't work).
As others said, there is no perfect SR systems, variety is the key.

A facility platform should have
-several, all motorized, widefield microscopes with deconvolution software
(most of our user coming for a "pretty image" are advised to try first a
good  acquisition  on a widefield (meeting Nyquist sampling requirements)
combined with deconvolution;
-at least one fast-live imaging widefield microscope (imaging less that
30ms),
-a TIRF (where you can change the angle to allow "dirty TIRF"/HALO/VAEM,
very useful when you need to image , plant cells a day, embryos and other
day and finally yeast
-a couple of confocals;
-1 or 2 SR scopes. From our experience, people likes to go on the AiryScan,
because, the increase in resolution is good enough for most of our users,
without preparing a new slide.


Have a great day
Chloë


*Chloë van Oostende, PhD*
Chair,* Plant Science* session
Canadian Microscopy and Cytometry Symposium 2017

Facility manager- Senior Microscopy specialist
*Cell Biology and Image Acquisition Core Facility*
Faculty of Medicine
University of Ottawa
RGN 3171
451 Smyth Road
Ottawa, ON K1H 8M5
Office: 613-562-5800 ext: 8376

http://www.chloevanoostende.net
CBIA core facility
<http://www.med.uottawa.ca/research/corelabs/CoreLabs_CBIA/eng/>

2017-08-25 6:49 GMT-04:00 Ferhan A <[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.
> *****
>
> Dear list members,
> Thank you very much for your on-list and off-list comments, help and
> suggestions.They were very useful and will help us a lot during decision
> making.
> I also thank Jacqui for starting this discussion.
> Have a nice weekend!
> Ferhan
>
> Ferhan Ayaydin, Ph.D.
>
> Cellular Imaging Laboratory
>
> Biological Research Centre
>
> Hungarian Academy of Sciences,
>
> Szeged, Hungary
>
> On Thu, Aug 24, 2017 at 4:04 AM, Jacqui Ross <[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 everyone,
> >
> > I'm enjoying the current conversation on super-resolution, especially
> > because we are currently investigating options for purchasing a
> > super-resolution microscope for our core facility. As you are all aware,
> > there are a number of different technologies, which all have pros/cons
> > depending on the specimen/labelling. We are currently considering STED,
> SML
> > (STORM/PALM), SIM, Airyscan, etc.
> >
> > I am looking for feedback from people working in core facilities like
> ours
> > where a large variety of research interests are supported, e.g. cultured
> > cells/tissue sections, plant/animal/bacteria, etc. Generally, our
> approach
> > is to train users to work independently rather than carrying out work for
> > people. I have already had a few conversations with colleagues in
> Australia
> > which have been very useful.
> >
> > If you work in a core facility and have one or more super-resolution
> > microscopes, I would love to hear from you. In particular, I would like
> to
> > know how often the super-resolution microscopes are used in comparison to
> > confocal for example. It would also be helpful to know whether
> researchers
> > expectations of the technology are being met and what level of support is
> > needed both from facility staff and from service engineers. Advice on
> data
> > processing requirements and/or problems would also be of interest. We are
> > consulting with our researchers via a survey and our feeling is that many
> > of our respondents are not realistic about what might be required in
> order
> > to get useful data. Of course everyone wants the highest possible
> > resolution, z stack capability, >2 fluorophores and live cell imaging.
> >
> > If you have more than one super-resolution system, which is more popular
> > and why? This purchase will require significant investment by our
> > institution and will require a business case so we want to make sure that
> > we make the best decision for our facility.
> >
> > Off list replies are welcomed and will be kept confidential to the
> members
> > of our working group if necessary. If you would prefer to provide your
> > views via telephone/skype, I'm happy to speak to you at a time that suits
> > you.
> >
> > Kind regards,
> >
> > Jacqui
> >
> >
> > Jacqueline Ross
> > Biomedical Imaging Microscopist
> > Biomedical Imaging Research Unit (BIRU)
> > School of Medical Sciences
> > Faculty of Medical & Health Sciences
> > The University of Auckland
> > Private Bag 92019
> > Auckland 1142, NEW ZEALAND
> >
> > Telephone: Ext 87438; DDI:  [phone]
> >
> > Website:    http://www.auckland.ac.nz/biru
> >
>

[Jacqueline Ross]

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*****

Dear listserv people,

Thanks very much to everyone who has replied to my enquiry so far both on the listserv and off-list. It was wonderful to see all of these replies coming into my Inbox over the weekend.

These comments are really helpful and I really appreciate the time that you have taken to reply. You have given us plenty to think about.

If anyone else still has thoughts to share (on/off list), then I am very happy to hear more.

Kind regards,

Jacqui

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Chloe van Oostende
Sent: Saturday, 26 August 2017 5:49 a.m.
To: [hidden email]
Subject: Re: Super-resolution microscopy - feedback wanted from staff at core facilities

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Hi,
Being a plant biologist who is working with numerous sample during my microscope facility experience (worms, yeast, human cells, tissue slices), I can state with a strong confidence, that a super-resolution (SR) microscope is under used.
We currently have a STORM, SIM, and STED. All under used because of different reasons: time consuming to have the perfect slide to be imaged, optimisation, time to get familiar with a software. Slides for STORM has to have a low staining, STED requires high staining, SIM works well on a restricted z-depth (multilayer of cells won't work).
As others said, there is no perfect SR systems, variety is the key.

A facility platform should have
-several, all motorized, widefield microscopes with deconvolution software (most of our user coming for a "pretty image" are advised to try first a good  acquisition  on a widefield (meeting Nyquist sampling requirements) combined with deconvolution; -at least one fast-live imaging widefield microscope (imaging less that 30ms), -a TIRF (where you can change the angle to allow "dirty TIRF"/HALO/VAEM, very useful when you need to image , plant cells a day, embryos and other day and finally yeast -a couple of confocals;
-1 or 2 SR scopes. From our experience, people likes to go on the AiryScan, because, the increase in resolution is good enough for most of our users, without preparing a new slide.


Have a great day
Chloë


*Chloë van Oostende, PhD*
Chair,* Plant Science* session
Canadian Microscopy and Cytometry Symposium 2017

Facility manager- Senior Microscopy specialist *Cell Biology and Image Acquisition Core Facility* Faculty of Medicine University of Ottawa RGN 3171
451 Smyth Road
Ottawa, ON K1H 8M5
Office: 613-562-5800 ext: 8376

http://www.chloevanoostende.net
CBIA core facility
<http://www.med.uottawa.ca/research/corelabs/CoreLabs_CBIA/eng/>

2017-08-25 6:49 GMT-04:00 Ferhan A <[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.
> *****
>
> Dear list members,
> Thank you very much for your on-list and off-list comments, help and
> suggestions.They were very useful and will help us a lot during
> decision making.
> I also thank Jacqui for starting this discussion.
> Have a nice weekend!
> Ferhan
>
> Ferhan Ayaydin, Ph.D.
>
> Cellular Imaging Laboratory
>
> Biological Research Centre
>
> Hungarian Academy of Sciences,
>
> Szeged, Hungary
>
> On Thu, Aug 24, 2017 at 4:04 AM, Jacqui Ross
> <[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 everyone,
> >
> > I'm enjoying the current conversation on super-resolution,
> > especially because we are currently investigating options for
> > purchasing a super-resolution microscope for our core facility. As
> > you are all aware, there are a number of different technologies,
> > which all have pros/cons depending on the specimen/labelling. We are
> > currently considering STED,
> SML
> > (STORM/PALM), SIM, Airyscan, etc.
> >
> > I am looking for feedback from people working in core facilities
> > like
> ours
> > where a large variety of research interests are supported, e.g.
> > cultured cells/tissue sections, plant/animal/bacteria, etc.
> > Generally, our
> approach
> > is to train users to work independently rather than carrying out
> > work for people. I have already had a few conversations with
> > colleagues in
> Australia
> > which have been very useful.
> >
> > If you work in a core facility and have one or more super-resolution
> > microscopes, I would love to hear from you. In particular, I would
> > like
> to
> > know how often the super-resolution microscopes are used in
> > comparison to confocal for example. It would also be helpful to know
> > whether
> researchers
> > expectations of the technology are being met and what level of
> > support is needed both from facility staff and from service
> > engineers. Advice on
> data
> > processing requirements and/or problems would also be of interest.
> > We are consulting with our researchers via a survey and our feeling
> > is that many of our respondents are not realistic about what might
> > be required in
> order
> > to get useful data. Of course everyone wants the highest possible
> > resolution, z stack capability, >2 fluorophores and live cell imaging.
> >
> > If you have more than one super-resolution system, which is more
> > popular and why? This purchase will require significant investment
> > by our institution and will require a business case so we want to
> > make sure that we make the best decision for our facility.
> >
> > Off list replies are welcomed and will be kept confidential to the
> members
> > of our working group if necessary. If you would prefer to provide
> > your views via telephone/skype, I'm happy to speak to you at a time
> > that suits you.
> >
> > Kind regards,
> >
> > Jacqui
> >
> >
> > Jacqueline Ross
> > Biomedical Imaging Microscopist
> > Biomedical Imaging Research Unit (BIRU) School of Medical Sciences
> > Faculty of Medical & Health Sciences The University of Auckland
> > Private Bag 92019 Auckland 1142, NEW ZEALAND
> >
> > Telephone: Ext 87438; DDI:  [phone]
> >
> > Website:    http://www.auckland.ac.nz/biru
> >
>

[Nbourg]

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----------------------- Commercial Response -------------------------

 

Dear all,

 

This discussion reminds me a lot my PhD work in the group of Sandrine Lévêque-Fort (ISMO, Orsay, France) during which I developed a new 3D SMLM technique (see http://www.nature.com/nphoton/journal/v9/n9/full/nphoton.2015.132.html).

 

We quickly realized that optics was only 1/3 of the problem as you also need to master chemistry (i.e. buffer) and software.

 

This is why we founded Abbelight (www.abbelight.com) : make homemade accessible to all. Therefore we have developed:

 

dSTORM super resolution buffer, easy to use and compatible with many fluorophores

Software for data acquisition and analysis (2D and 3D real-time reconstruction, drift compensation, 3D viewer and many more...) compatible with many systems (both commercial and home built)

Add-on module to upgrade an inverted microscope into a 3D nanoscope (DONALD technology)

Full super resolution packages (microscope, video camera, lasers, TIRF module, 3D super resolution module)

 

Either you want to acquire a full system, build it or add some features to your existing one, we have a solution.

 

We have now gained trust of several renowned research institutes especially thanks to our dSTORM buffer, including Imperial College of London, Cambridge University, Pierre and Marie Curie University, Curie Institute, University of Pennsylvania and many more.

 

Please let me know if you are interested so we can talk about it.

 

Dr. Nicolas Bourg

0033 6 38 82 82 05

[hidden email]

www.abbelight.com