LCI microscopy course, vintage 2021 :)

The Live Cell imaging facility will run again its intensive light microscopy course in Jan-Feb 2021.

The schedule can be found here.

As usual, all lectures will be publicly broadcasted live. So if you think that a lecture could be useful for you, you are welcome to listen, without registration, by following this link.

Due to the current pandemics, only registered students will be allowed in the lecture room.

The course comprises lectures, workshops, imaging of your own sample, demos… It will run 26 Jan-12 Feb, 3 days/week (tues-thurs) 9:00-17:15.

There are only 16 spots. The course counts for 6 credit points.
The rest of the time (the course counts for 4 weeks) is used for home assignments. Mondays and Fridays will also be used for individual workshops where we image your own sample.

The application process is open now until the 16th of November. Please read carefully the eligibility criteria. You will need to sent me some images of your sample at the time of application. This course is only open to people who already have some experience of fluorescence microscopy.

Microscopy: improve your imaging skills – from sample preparation to image analysis

At the end of the course, the participants will be able to:
1- Describe the difference between wide field, confocal and light sheet microscopes as well as the different types of confocal microscopes and choose which system is most suited to their experiments
2- Pick the best combination of fluorophores for their experiment by matching their spectra with the microscope light source and filters, identify and eliminate bleed-through and cross-excitation problems
3- Explain objective specifications and limitations and choose the appropriate objective for their own experiments
4- Describe how to fix, mount and handle their sample in a way that is optimal for imaging
5- Find their sample and the area of interest without bleaching it
6- Adjust the condenser for proper DIC imaging (Koehlering)
7- Explain how to set the following parameters on a wide field, a confocal or a light sheet system to best match the requirements of their sample and reliably answer their scientific question: resolution, pixel size, averaging, scan speed, illumination power, detector gain and offset, camera readout rate, exposure time and camera binning
8- Explain which applications require a hardware or a software autofocus, a spectral detector, a resonance scanner, two-photon or super resolution microscopy
9- Explain the advantages in using the automation of a microscope system to collect multidimensional data
10- Explain how to deal with images before publication in scientific journals
11- Describe the imaging requirements for automated image analysis
12- Run an image analysis pipeline on freeware (ImageJ/FIJI, Cell Profiler) designed for their own images and scientific question.

Please spread the word to your colleagues.

We are looking forward to meeting you and your sample! 😃

Tweety turned into a power machine!

Our dear Tweety microscope, which was simplest and cutest of all the LCI systems, has muted into our most sophisticated power machine!

On the 24th at 10, we will run an online demo (link below) to show what our upgraded Tweety can deliver:

  • Much larger field of view (from 18 mm diagonal to 25 mm)
  • Upgraded single point confocal on the left side
  • Resonant scanner with 1024×1024 pixels (compared to 512×512 on our other resonant scanners), still the same speed (30 fps) and improved low noise
  • Spinning disk confocal on the right side with bypass to image wide field
  • 2 very sensitive cameras on the right side: one with the very large field of view and 11um pixels for best sensitivity and one with the normal field of view and 6.45 pixels for best resolution
  • Our great Primo is still on the back of the microscope to allow micropatterning of proteins at the bottom of a dish or micromanufacturing of wells in the shape/pattern of your choice
  • 2 wonderful silicon immersion objectives specialized for tissue imaging with automatic correction ring: 20x/1.05 and 40x/1.25

After the demo, the LCI users who have already been trained on our widefield systems can get access to Tweety for free after a mandatory short training.

Please add the demo in your calendar and make sure to test the link ahead of the meeting.

Link to the Zoom Meeting on the 24th at 10am: https://ki-se.zoom.us/j/7302561100

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

Our course starts tomorrow! 😀

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)

Seminar on how to directly label your primary antibodies: Skip the secondary, part 3!

The LCI will host a seminar about GlyCLICK, a new way to directly label primary antibodies and stop using secondaries: 6th of November at 13:00 in the Lipid seminar room in Neo, KI Flemingsberg. The seminar will be broadcasted live. Here is how to find us and here how to follow the seminar online.

Everyone agrees that it would be great to be able to label our samples without using secondary antibodies.

  • less animals killed
  • shorter and cheaper protocols
  • no problem with isotype cross-reaction
  • no problem with secondary species when using many antibodies at once

There are many kits to label primary antibodies directly with fluorophores. The main disadvantage compared to primary/secondary stainings is that direct labels are often weaker because the final primary/fluorophore ratio is too low. Using an amplification method like TSA (Tyramide Signal Amplification) leads to a loss of resolution.

Over the years users at the LCI have tried this kit, this kit and this one. They gave mixed results depending on the antibody but we keep looking! Come to the LCI seminar about a new direct labelling technique that uses Click chemistry.

Apply now to the LCI microscopy course 2020 :)

It is now time to register to the LCI intensive microscopy course (Jan/Feb 2020). Check out the course schedule.

Loads of fun workshops, informative lectures, intense discussions and our popular Student Imaging Challenge workshop where students get direct feedback on how to improve their own sample preparation/experimental design.

We always run two courses in parallel:

  • the full course (#2870, 6 points, apply here) where students attend all activities
  • the theory only course (#2871, 4.5 points, apply here) for students who only attend the lectures

As usual, all lectures are public and broadcasted live so you are welcome to just show up (how to find us) or watch remotely (how to connect) without registration.  Check the program as it may be updated in case of (unlikely) last minute changes.

We welcome your feedback about the quality of the webinar and the content of the lectures.

Party time at the LCI! :D

On the 10th of October, the Live Cell Imaging facility will have an open house and a little party to celebrate loads of great stuff:

  • The LCI turned 10 years old this year! 😊
  • We got a wonderful light sheet system earlier this year. It is high time to give it a name and splash it with a bit of champagne!
  • Gisele Miranda from Scilife/BII has joined the LCI team to help our users with image analysis
  • We got a great server/analysis capacity set up by the KI IT department

Wow! What a year! 😃

Please come and celebrate with us! If you do not know what our microscopy facility has to offer, it is time to be curious and pay us a visit.

  • Open house: drop in between 9:00-11:00
  • Baptising of the light sheet system and celebration of Gisele and our shiny new server: at 11:00

All this will happen on the 10th of October (next Thursday) at the LCI facility, on the 7th floor of Neo at KI Flemingsberg. Here you can see how to find us.

Please help us spread the news! 😊

Call4Help: fast-track help with your image analysis project!

BII (BioImage Informatics, the great image analysis at SciLife Uppsala) and the LCI facility will run a new Call4Help on the 4th of September.

Anyone who is stuck with image analysis and wishes for quick help to build a pipeline should apply. You don’t have to acquire the images at the LCI. Anyone can apply.

How does it work? You first upload your images and a little explanation text. A few days later, we all meet virtually in a Zoom chatroom for a quick (30 min-1 h) online session. You get comments, suggestions and help with building a Fiji or CellProfiler analysis pipeline tailor-made for your images.

If you are interested, click on the link below to apply:

BioImage Informatics

Super-Resolution spinning disk demo at the LCI!

Dear microscope freaks

How would you like to run some gentle live sample imaging with a 60x objective with:

  • an xy resolution of 120 nm without software tricks (or even better after deconvolution),
  • the great contrast of a true confocal,
  • 82 frames per second,
  • or decide to bypass everything, go widefield and image at 100 frames per second with a super large field of view (220×220 um)?

Sounds good to me! 🙂

For the next 2 weeks you can do that with the new toy on demo at the LCI facility!

The beast is a new sort of spinning disk confocal and is called SoRa (Super-Resolution Optical Reassignment). It is a collaboration between Nikon and Yokogawa.

We even have 2 cameras to compare (Prime95B and BSI from Photometrics).

Oliver Garner from Bergman Labora will give a short online presentation of how SoRa works on Monday (29th) at 13:00. The presentation is done remotely and broadcasted live. You can join the audience from the comfort of your office chair by following the instructions here (please try beforehand to make sure all works).

Interested in trying it? Please contact us.

A dream!!

Call4Help: The image analysis help you have always dreamt of, totally for free!!! ???

After the success of the previous Call4Help session in February, BII (BioImage Informatics, the great image analysis at SciLife Uppsala) and your favorite microscopy facility (we hope) will run a new Call4Help next Tuesday 2nd of April.

Anyone who is stuck with image analysis and wishes for quick help can apply.

These are 100% online sessions (we ‘meet’ in a Zoom chat room) where you submit your images and a little explanation text in advance and you get suggestions for 30 min and an analysis pipeline all done for you (Fiji, CellProfiler, Ilastik, QuPath, KNIME)!

If you are interested, please apply as soon as possible (sorry for the late announcement).

Here is how to apply: https://www.scilifelab.se/facilities/bioimage-informatics/

Image Analysis Call4help on 14th of feb!

The image analysis help you have always dreamt of, totally for free!!! ???

Deadline 6th of feb!

First SciLifeLab BioImage Informatics call4help zoom session

On the 14th February 2019, 10:00 – 12:00 the SciLifeLab BioImage Informatics Facility is organizing an online support session on BioImage Analysis (“call4help” session).

How does it work?

Each selected participant gets a 15 min slot. You have 5 min to present your problem. It is mainly about showing the scientific context, representative input images, describing desired output measurements, and giving a brief statement about what was done before. The remaining 10 min we will brainstorm together on the possible solution(s), discuss tools, techniques, etc.

The Bioimage Informatics Facility can provide further support, if needed. We are happy to announce that Sylvie Le Guyader, Live Cell Imaging Facility, KI, Huddinge, is joining the session as microscopy expert.

How can I participate?

Step1:

Prepare a presentation – use this template for your presentation:

Step2:

a) Upload your presentation to this google drive:

b) AND send an e-mail to anna.klemm@it.uu.se until 6th February 2019.

Step 3:

Join this zoom-session.

If your problem is selected for presentation, you will get a time-slot of 15 minutes within the session (14th February 2019, 10:00 – 12:00). If not, you are welcome to join the discussion anyway!

What are the Deadlines

Submission of a problem: until 6th February 2019

Notification about participation: 8th February 2019

Session: 14th February 2019, 10:00 – 12:00

Participants

Petter Ranefall

Kevin Smith

Anna Klemm

Sylvie Le Guyader

Call4Help format

The call4help format was initially developed by ScopeM, ETH Zürich.