Tag: Imaging

Live demo of the Crest spinning disk with DeepSIM

9th of September, 10-12am: Live/zoom demo of the latest microscope acquired by the LCI facility: Crest spinning disk with DeepSIM (new Orion)

  • Ti2 microscope
  • Crest spinning disk (identical to the one on Tweety but with slightly different wavelengths)
  • DeepSIM: SIM is a fast super-resolution technique which is gentle for the sample (less bleaching, less photodamage for live samples) and offers double the resolution of a confocal microscope (down to 120nm resolution), even thick in samples. It is flexible and can be used with any objectives, even low magnification air objectives.
  • No specific sample preparation requirements
  • More information here

For the best experience, join the demo in person at the LCI facility (please apply by replying to this email) but if you cannot make it, you can also listen on Zoom.

The system has been purchased by the LCI so it is here to stay! šŸ™‚

Please let us know if you want to join the demo in person and mention if you would like to try imaging your own sample with DeepSIM.

mCherry-XL: brigther, more stable and with better spectra!

mCherry is a very popular red fluorescent protein. However it has several disadvantages:

  • It is shifted towards far red (ex peak 585 nm) so it often is not imaged optimally with the illumination sources and filters commonly available.
  • It bleaches fast

mCherry has now been evolved into mCherry-XL with several improvements:

  • This variant is shifted back towards green (ex peak 560 nm) therefore being very well excited with popular 561nm lasers.
  • It is 3 times brighter than mCherry
  • There is also a clear improvement in the lifetime for FLIM
  • Together the 2 points above means that less excitation power is required so it should help with the bleaching problem

Here is the paper. Therefore you should consider mCherry-XL for your future tagging with red fluorescence proteins.

Super resolution STED event at Scilife in March

Nordic superresolution microscopy facility staff/researchers: STED

March 15th and 16th at 9-12 am. CONTACT: Hans Blom (hblom@kth.se)

Mar 15, 2022 9:00am

  • 9:00-9:45am – BNMI info / STED introĀ [Hans Blom & Daniel Smeets (Leica)]
  • 9:45-10:15am –Ā STED sample preppingĀ [Ulf Schwarz et al. (Leica)]
  • Break
  • 10:30-11:00am – Input from a Nordic STED facility [Jonathan R. Brewer (DaMBIC)]
  • 11:00-11:45am –Ā STED demosĀ [Luis Alvarez/Ulf Schwarz et al. (Leica)]
  • 11:45-12:00am – Extra discussion time

https://kth-se.zoom.us/j/69376340781

Mar 16, 2022 9:00am

  • 9:00-9:45am – Live-cell STED [Giovanna Coceano (KTH Stockholm)]
  • 9:45-10:15am – Remote STED [Marko Lampe (ALMF/EMBL)]
  • Break
  • 10:30-11:00am – STED-FCS [Erdinc Sezgin (KI Stockholm)]
  • 11:00-11:45am – Advanced STED demosĀ [Luis Alvarez/Ulf Schwarz et al. (Leica)]
  • 11:45-12:00am – Extra discussion time

https://kth-se.zoom.us/j/67902191893

The LCI microscopy course 2022 is now open for registrations!

It is now time to apply to the intensive LCI microscopy course Jan/Feb 2022: Microscopy: improve your imaging skills – from sample preparation to image analysis

The aim for this course is to improve the microscopy skills of students and researchers who have already used a microscope to acquire digital images of fluorescent samples but feel that their knowledge is limited.

All the lectures at the LCI microscopy course will also broadcasted live online, free of charge and there is no need to register.

All details about the course including course schedule, how to apply, and how to follow the lectures are found here.

Scroll down to read the kind testimonies of our dear students! šŸ˜Š

Hope you enjoy the LCI facility microscopy course 2022!

Great webinars about sample preparation at Scilife

Hans Blom from the Advanced Light Microscopy facility at Scilife organises next week (10th and 1tth of June) 3 sessions of great seminars:

I especially recommend the CUBIC seminar to anyone who wants to image samples thicker than 100 um. This will teach you how to treat your sample to make it fully transparent so you can image through many cm of tissue!

šŸ™‚

Super bright green fluorescent protein

Fluorophores are evolving fast! Here is a paper about a bunch of new fluorophores isolated from the jelly fish Aequoria. This includes the brightest fluorophore ever isolated: a new green fluorescent protein called AausFP1 that is almost 5 times brigther than GFP! Respect! šŸ™‚

A microscopy-related job at EMBL Heidelberg with EuroBioImaging

Euro-BioImaging Bio-Hub at EMBL Heidelberg is looking for the Euro-BioImaging Industry Board (EBIB) Coordinator who will work in close collaboration with EBIB Chair and EBIB members located across Europe as well as with the Euro-BioImaging Hub Office.

The Euro-BioImaging Industry Board (EBIB) is currently comprised of thirteen different companies across the imaging field. In the EBIB, all companies work together as a single, professional entity, setting their own goals in order to proactively drive the interaction between the imaging industry on the one hand and European researchers, funders and the imaging facilities linked with the European research infrastructure, Euro-BioImagingĀ on the other hand. In October 2019, the European Commission has officially established Euro-BioImaging ā€“ which provides life scientists with open access to a broad range of technologies and resources in biological and biomedical imaging ā€“ as a European Research Infrastructure Consortium (ERIC). The post holder will report to the Director Euro-BioImaging Bio-Hub at EMBL and the EBIB Chair. The responsibilities of the position include among others:

  • Proactively seeking opportunities for EBIB members to engage and collaborate with Euro-BioImaging Nodes, users and national networks;
  • Representing the EBIB at international events in a professional and politically astute manner;
  • Strategically prepare and operationally manage EBIB-driven meetings and workshops to understand the needs of the imaging community;
  • Coordinating “One Voice” activities to the political community at national and European level (European Commission, Brussels) in order to increase the imaging market;
  • Developing communication tools and activities to boost EBIBā€™s visibility in order to increase the number of industry members and strengthen EBIBā€™s presence in the scientific and political landscape;
  • Keep up-to-date with imaging-associated initiatives and provide EBIB with tailored insight into European research trends and culture changes;
  • Coordinating industry participation in Euro-BioImaging EU-wide training programs;

If you are interested please find hereĀ the complete job description and link for online submission of your application.

Please distribute also to your friends and colleagues, who might be interested in this job posting. Thank you!

Kind regards,

Antje Keppler, PhD

Interim Section Director

Euro-BioImaging Bio-Hub

Global BioImaging Coordinator

EMBL Meyerhofstr. 1, DE-69117 Heidelberg Tel. +49 6221 387 8847

One-step non-toxic clearing protocol

Do you know that clearing is not just about light sheet microscopy? Even if you have done your job well and your sample is directly on the coverslip (not on the slide), as soon as your sample is thicker than 10 um (1 cell diameter), you will see the effect of the refraction index mismatch.

What is that? Your sample and the mounting medium around it have a certain refraction index (or likely several). The objective you are using is designed for a certain refraction index (e.g. air, water or oil). If these refraction indices do not match what happens? as soon as you image a tiny bit away from the coverslip, the sample will look elongated, the intensity and contrast will drop very fast.

Sounds familiar? If yes, changing your mounting medium to match the objective will solve the problem. It works for light sheet but it also works for wide field or confocal imaging! Just change your mounting medium and you will see an enormous difference!

Here is an article describing a one-step clearing protocol. This basically is about using a different mounting medium. Easy, cheap and non-toxic! Give it a try!

Have a look at this post for more info.

Broadcasted lectures from the LCI microscopy course and private demos of light sheet and cameras

Our course starts tomorrow! šŸ˜€

Target audience:

The aim for this course is to improve the microscopy skills of students and researchers who have already used a microscope to acquire digital images of fluorescent samples and want to improve their skills.

Registrations are closed but all lectures are open to everyone without registration.

  • The schedule and details of the venue are here.
  • All lectures are also available online live. The link and instructions to watch are here.
  • Make sure you check the schedule in case of last minutes changes.

If you are in Sweden, you are welcome to try some of the equipment on demo with your own sample.

To book at timeslot, please contact the responsible person directly.

  • Light sheet microscope from M2Lasers: Valentina Loschiavo Valentina.Loschiavo(at)m2lasers.com
    • Fast imaging of large sample
    • Overview function to navigate in the sample and find the region of interest
    • 800x800um field of view with 1um min resolution
    • Any immersion media
    • Sample size up to centimetres
  • Wide Field microscope from Nikon with 3 different Andor cameras: Oliver Garner (oliver.garner(at)bergmanlabora.se)
    • Nikon Ti2 microscope with 4 times larger field of view
    • A front illuminated sCMOS camera: Good sensitivity and resolution, great speed, but a greyish background (Andor Zyla 4.2)
    • A back-illuminated EM-CCD camera: highly sensitive camera with very dark background, but lower resolution and lower speed (Andor 897U)
    • A back-illuminated sCMOS camera: same sensitivity and low background as an EM-CCD but better resolution and speed (Andor Sona)

Fluorophores that blink for STORM without any special buffer

Sounds good! No more buffer that stops working after one hour of imaging!

This company sells SaraFluor650 secondary antibodies. This fluorophore seems to be a natural blinker which does not need to reducing environment. Apparently it doesn’t need very bright lasers either!

Apparently they have also developed a green variant. So you can run 2-colour direct STORM on your favorite TIRF system! šŸ˜€

They also sell pH sensors and more so check their website and if you try, write a comment to let us know how things worked!

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