Best practice to prepare figures with microscopy images

Here you can find information about a very interesting talk on the 5th of March (7pm, Swedish time) about the recommended practice to prepare figures containing microscopy images.

This talk is based on a recent Nature communications article Community-developed checklists for publishing images and image analysis.

The LCI open microscopy course starts on Monday! :)

Dear all

It is my pleasure to invite you to follow the LCI facility microscopy course Microscopy: improve your imaging skills – from sample preparation to image analysis.

The course starts next Monday (29 Jan) and runs until the 16th of February.

As usual, all the course lectures are broadcasted live on Zoom. It is free of charge and you do not need to register.

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 more knowledge could help them.

Here is a selection of what we will talk about:

  • Optics and image formation,
  • Anatomy of a microscope
  • Objectives and refraction index
  • Cameras and detectors, Noise and background, Bit depth and saturation
  • Sample preparation, Immunostaining, Clearing and expansion
  • Resolution and contrast, Nyquist sampling, Microscope settings
  • Data handling, OMERO.figure, Requirements for image analysis, Colocalization
  • Image processing and analysis

Check our course webpage to see the course schedule (Broadcasted activities are in blue) and the Zoom link. Scroll down to read the kind testimonies of our dear students! 🙂

Here is the course syllabus.

For those who are on a far away time zome, we record all the live lectures and post them every evening on the LCI facility YouTube channel! 🙂

We hope that you will enjoy the LCI facility microscopy course!

Kindly forward to anyone who might be interested.

The LCI team

A few spots left at the LCI microscopy course 29/01-16/02 2024

There are a few spots left for the LCI core facility light microscopy course (29 Jan – 17 Feb 2024): Microscopy: improve your imaging skills – from sample preparation to image analysis’ (6 credits).

This course is completely unique in that it is a highly hands-on, but your hands will be on your own microscope and own sample. The course runs completely remotely! 😊

  • To see how the course can help your microscopy project, check the course webpage. Look at the course schedule 2024 and the alumni testimonies!
  • Also check the course syllabus to see the eligibility criteria. Also read what you will learn in the Course content and Intended Learning Outcomes sections.
  • If you cannot apply to the course, you can anyway follow any of the lectures (in blue on the schedule) as they will be publicly broadcasted live on Zoom and accessible to anyone without registration. The schedule and zoom link are available on the course page.

The purpose of the LCI facility microscopy course is to provide PhD students, researchers and core facility staff who have some prior experience of microscopy with enough theoretical and practical knowledge about their OWN sample and their OWN microscope, to enable them to:

1) assess and improve their sample so that it becomes suitable for data extraction from fluorescence images,

2) make best use of the hardware available in their lab/facility,

3) fully understand the acquisition parameters they need to set in their own microscope software,

4) design their experiment from scientific question to image analysis using a strong knowledge base.

The aim is to provide you with tools to acquire on ANY wide field, confocal or light sheet microscope, images of your samples that reliably answer your scientific question.

The course is free of charge. Contact us (LiveCellImaging@ki.se) for enquiries.

Spatial transcriptomics in Flemingsberg!

Spatial transcriptomics techniques are booming! It is now possible to identify where RNAs are located within a tissue! At the LCI facility, we routinely image samples labelled with RNAscope. If you use RNAscope, you might be interested in this workshop where you will learn how to label thick samples with RNAscope, and this symposium about automated RNAscope.

Additionally, we can now offer the 10x Genomics Visium technology on the South Campus, as a collaboration between 4 core facilities in Flemingsberg: LCI, FENO, SICOF and BEA core facilities!

Here is the workflow:

  1. We discuss your project and advise you for the preparation of your sample.
  2. You cut your sample and optimize the antibody or H/E staining yourself, under the guidance if the LCI staff to best preserve the RNAs.
  3. You image your tissue sections at the LCI facility.
  4. You transfer your section to SICOF who tags the RNAs and amplifies the library, using the 10x Genomics Visium technology.
  5. The library goes to BEA for sequencing.
  • Section size: max 6.5×6.5 mm or max 11×11 mm
  • Human or mouse
  • Paraffin-embedded or fresh-frozen
  • Labelled with H/E or fluorescent antibodies

Come and talk to us about your Spatial Transcriptomics project!

Nordic Microscopy Symposium – 3rd-5th October

We are delighted to invite you to the Nordic microscopy symposium at the Live Cell Imaging core facility, Nikon Centre of Excellence at Karolinska Institutet, organised by BergmanLabora, Ramcon Denmark and Inter Instrument AS.

  • The symposium will highlight the great research done in the Nordics where microscopy is a key tool.
  • New exciting equipment from Nikon will be showcased including the AX/NSPARC super-resolution point confocal and the CrEST DeepSIM super-resolution spinning disk confocal.
  • We will also be launching the new Nikon Eclipse Ji benchtop assay instrument and demo the simpler CrEST Cicero spinning disk confocal.
  • The capability of these systems will be demonstrated during public sessions (3rd of October) and in private sessions (on 4th and 5th October) where you are welcome to bring your own samples.

Venue: Neo, Flemingsberg Campus, Karolinska Institute: Erna Möller Lecture hall (Symposium), Live Cell Imaging (LCI) core facility (Demos)

3rd October

    • 1000-1145       Public demo: Nikon AX R NSPARC super-resolution point-scanning confocal, CrEST X-Light V3 super-resolution spinning disk confocal, CrEST Cicero compact spinning disk
    • 1000-1145       Open house at the Live Cell Imaging core facility (Drop in. Contact Sylvie Le Guyader)
    • 1200-1300       Lunch (requires pre-registration)
    • 1300-1305       Welcome
    • 1305-1345       Zuzana Kadlecova (Cambridge Institute for Medical Research – UK): Lights! Camera! Uptake! Zooming in on Endocytosis with Quantitative TIRF-SIM Analysis.
    • 1345-1410       Nicoline Dorothea Daugaard (Dept. Biochemistry and Molecular Biology, SDU – Denmark): Tracking T cells and their interactions with cancer cells in 3D using NIS-Elements General Analysis (GA3)
    • 1410-1440       LCI short talks:
      • Chiara Annunziata (Karolinska Institute – Sweden): A pipeline for the multiparametric assessment of the anti ageing effects of candidate molecules
      • Andrea Coschiera (Karolinska Institute – Sweden): Primary cilia promote the differentiation of human neurons through the WNT signaling pathway
      • Natalie Geyer (Karolinska Institute – Sweden): Multiplex RNA in situ hybridisation for liver parenchyma characterization in a metastatic context
    • 1440-1500       Coffee
    • 1500-1525       Technical talk (Nikon): Microscopy Simplified, Nordic Launch of Nikon’s new ECLIPSE Ji
    • 1525-1540       Staffan Strömblad (Karolinska Institute – Sweden): Multi-site assessment of reproducibility in high-content cell migration imaging data
    • 1540-1605       Pieta Mattila (University of Turku – Finland): Advanced imaging methods to visualize lymphocyte activation

4th and 5th October

Registration is mandatory and can be done via the following site: Nordic Microscopy Symposium – BergmanLabora.

Please share this event with friends and colleagues who may also be interested in attending.

Clearing and expansion microscopy symposium and workshop sept 12-14

Two really nice symposia/workshop on clearing and expansion microscopy next week!

Nordic microscopy symposium – Save the date!

Please join the Nordic microscopy symposium on the 3rd of October at the Live Cell Imaging core facility, Nikon Centre of Excellence at Karolinska Institutet. The symposium is organised by BergmanLabora, Ramcon Denmark and Inter Instrument AS. Please save the date.

The event will highlight Nordic research where light microscopy is a key tool.

Several Nikon fast super resolution systems will also be available: Nikon AX confocal with NSPARC and Nikon/CrEST V3 spinning disk with DeepSIM.

Before and after the symposium, the capability of these systems will be demonstrated during public sessions (on the 3rd of October) and in private sessions (booking required) where you are welcome to bring your own samples.

Preliminary schedule:

Tuesday 3rd October

1000-1045     AX R NSPARC + CrEST V3 with DeepSIM public demonstration (in parallel)

1100-1145        AX R NSPARC + CrEST V3 with DeepSIM public demonstration (in parallel)

1200-1300     Lunch (requires pre-registration)

1300-1305     Welcome

1305-1345      *Keynote: TBC

1345-1410       *Nicoline Dorothea Daugaard (Dept. Biochemistry and Molecular Biology, SDU – Denmark):

Tracking T cells and their interactions with cancer cells in 3D using NIS-Elements General Analysis (GA3)

1410-1440       *LCI short talks:

Chiara Annunziata (Karolinska Institutet – Sweden)

Andrea Coschiera (Karolinska Institutet – Sweden)

Natalie Geyer (Karolinska Institutet – Sweden)

1440-1500     Coffee

1500-1525      *Super resolution system overview (Nikon): Title TBC

1525-1540      *Staffan Strömblad (Karolinska Institutet – Sweden): Multi-site assessment of reproducibility in high-content cell migration imaging data

1540-1615       *Pieta Mattila (University of Turku – Finland): Title TBC

Wednesday 4th October

0830-1400    AX R NSPARC + CrEST V3 with DeepSIM private demonstrations (bring your own sample)

Thursday 5th October

0830-1400    AX R NSPARC + CrEST V3 with DeepSIM private demonstrations (bring your own sample)

Venue (symposium): Erna Möller Lecture hall, Neo, Flemingsberg Campus, Karolinska Institutet

Venue (system demonstrations): Live Cell Imaging core facility, Neo, Flemingsberg Campus, Karolinska Institutet

Please share this event with colleagues who may be interested in attending.

Live broadcast of all public lectures the LCI microscopy course

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

Starting from next week, the Live Cell Imaging core facility at KI will run its yearly microscopy course.

All the course lectures will be broadcasted live on Zoom, free of charge and there is no need to register. On this page, you can find the course schedule (public activities are in blue) and the  Zoom link to join. Scroll down to read the student testimonies! 😊

30 Jan – 17 Feb 2023

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 more knowledge could help them.

The course covers the following topics:

  • Optics, image formation
  • Fluorescence, fluorophores
  • Bleedthrough
  • Anatomy of a microscope
  • Objectives and refraction index
  • Cameras and detectors
  • Noise and background, Bit depth and saturation
  • Multichannel imaging and spectral unmixing
  • Resolution and contrast
  • Sample preparation, Immunostaining
  • Nyquist sampling
  • Confocal and wide field settings
  • Speed, High throughput/content
  • Volume imaging, deconvolution
  • Clearing and expansion
  • Live cell imaging
  • Fourier
  • AI, Super Resolution microscopy
  • Colocalization
  • Data handling, OMERO.figure

Hope you enjoy the Live Cell Imaging core facility microscopy course! 😃

BII webinar about a great tool! TissUUmaps

The BioImage Informatics facility at Scilife organizes a new presentation on free and open-source tools: TissUUmaps, a browser-based tool for GPU-accelerated visualization and interactive exploration of millions of datapoints overlaying tissue samples.

Users can visualize markers and regions, explore spatial statistics and quantitative analyses of tissue morphology, and assess the quality of decoding in situ transcriptomics data. TissUUmaps provides instant multi-resolution image viewing, can be customized, shared, and also integrated in Jupyter Notebooks. TissUUmaps was created in collaboration between BIIF and the Wählby lab. You can read more about it and test the software on its web page: https://tissuumaps.github.io/
During the seminar, we will go through basic usage of TissUUmaps: installation, loading images, markers and regions, change visualization settings, and how to load / save / share projects. The webinar will be given by Christophe Avenel and will take place on March 3rd, 09:00-10:00 (instead of our normal Call4Help session). There will be time for questions and discussion, so we hope this event to be very interactive. Please register here.

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

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

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