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! ๐Ÿ™‚

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! ๐Ÿ™‚

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): โ€˜Microscopy: improve your imaging skills – from sample preparation to image analysisโ€™ (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! ๐Ÿ˜Š

  • 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!

Time to register to the LCI microscopy course!

Dear all

It is time to enroll to the yearly light microscopy course run by the Live Cell imaging core facility (29 Jan – 17 Feb 2024): โ€˜Microscopy: improve your imaging skills – from sample preparation to image analysisโ€™ (6 credits).

  • Check the course schedule 2024 and the syllabus to see the course content and alumni testimonies.
  • Postdocs or registered PhD students can apply to the course.
  • Please read carefully the eligibility criteria in the syllabus and note that the last registration date is the 15th of November.
  • 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 on the course page.

The purpose of this course is to enable PhD students and researchers who have already and recently used a microscope to acquire images of fluorescent samples, to improve their microscopy skills.

The course is NOT aimed at training people to use the LCI facility microscopes. The focus is instead on providing the students with enough theoretical and practical knowledge about their OWN sample and their OWN microscope, to enable them to:

1) prepare their sample and formulate their scientific question in a way that is suitable for data extraction from fluorescence images

2) properly use the hardware available in their lab/facility and 3) fully understand each parameter they need to set in the software in their lab/facility.

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

Please help us spread the word. ๐Ÿ˜ƒ

The need to improve microscopy trainings

The LCI facility has just published an article aboutย  โ€˜Improving light microscopy training routines with evidence-based educationโ€™. The article is now available in the Journal of Microscopy. We hope that this article will give useful tips to better design usersโ€™ training and improve their learning outcome. ๐Ÿ™‚

Here is the abstract:

The low reproducibility of scientific data published in articles has recently become a cause of concern in many scientific fields. Data involving light microscopy is no exception. The low awareness of researchers of the technologies they use in their research has been identified as one of the main causes of the problem. Potential solutions have hinted at the need to improve technological and methodological education within research.

Despite the pivotal role of microscopy core facilities in the education of researchers being well documented, facility staff (FS) often learn their trade on the job, without receiving themselves any structured education about the technology they teach others to use. Additionally, despite endorsing an important role at the highest level of education, most FS never receive any training in pedagogy, the field of research on teaching and learning methods.

In this article, we argue that the low level of awareness that researchers have of microscopy stems from a knowledge gap formed between them and microscopy FS during training routines. On the one hand, FS consider that their teaching task is to explain what is needed to produce reliable data. On the other, despite understanding what is being taught, researchers fail to learn the most challenging aspects of microscopy, those involving their judgement and reasoning. We suggest that the misunderstanding between FS and researchers is due to FS not being educated in pedagogy and thus often confusing understanding and learning.

To bridge this knowledge gap and improve the quality of the microscopy education available to researchers, we propose a paradigm shift where training staff at technological core facilities be acknowledged as full-fledged teachers and offered structured education not only in the technology they teach but also in pedagogy. We then suggest that training routines at facilities be upgraded to follow the principles of the Constructive Alignment pedagogical method. We give an example of how this can be applied to existing microscopy training routines. We also describe a model to define where the responsibility of FS in training researchers begins and ends.

This involves a major structural change where university staff involved in teaching research technologies themselves receive appropriate education. For this to be achieved, we advocate that funding agencies, universities, microscopy and core facility organisations mobilise resources of time and funding. Such changes may involve funding the creation and development of โ€˜Train-the-trainerโ€™ type of courses and giving incentives for FS to upgrade their technological and pedagogical knowledge, for example by including them in career paths. We believe that this paradigm shift is necessary to improve the level of microscopy education and ultimately the reproducibility of published data.

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

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

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

1200-1300ย ย ย ย  Lunch (requires pre-registration)

1300-1305ย ย ย ย  Welcome

1305-1345ย ย ย ย ย  *Keynote: TBC

1345-1410ย ย ย ย ย ย  *Nicoline Dorothea Daugaard (Dept. Biochemistry and Molecular Biology, SDU โ€“ Denmark):

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

1410-1440ย ย ย ย ย ย  *LCI short talks:

Chiara Annunziata (Karolinska Institutet โ€“ Sweden)

Andrea Coschiera (Karolinska Institutet โ€“ Sweden)

Natalie Geyer (Karolinska Institutet โ€“ Sweden)

1440-1500ย ย ย ย  Coffee

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

1525-1540ย ย ย ย ย  *Staffan Strรถmblad (Karolinska Institutet โ€“ Sweden): Multi-site assessment of reproducibility in high-content cell migration imaging data

1540-1615ย ย ย ย ย ย  *Pieta Mattila (University of Turku โ€“ Finland): Title TBC

Wednesday 4th October

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

Thursday 5th October

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

Venue (symposium): Erna Mรถller 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 ย Zoom link to join. Scroll down to read the student testimonies! ๐Ÿ˜Š

30 Jan โ€“ 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! ๐Ÿ˜ƒ

Registration to the LCI microscopy course 2023 closes on Monday!

It is time to enroll to the yearly light microscopy course run by the Live Cell imaging core facility (30 Jan – 17 Feb 2023): โ€˜Microscopy: improve your imaging skills – from sample preparation to image analysisโ€™ย (6 credits).

  • Check theย course scheduleย to see the course content and testimonies from alumni
  • If interested, you canย enrol as a student.ย Please read carefully the eligibility criteriaย and note thatย the last registration date is the 15th of November.

Additionally,ย all the lectures (in blue on the schedule) will be publicly broadcasted live on Zoom and accessible to anyone without registration. Even if you do not want to enrol as a student, you can have a look at the and listen to any lecture that triggers your interest. There is no need to register. The schedule and zoom link are on theย course page.

The purpose of this course is to enable PhD students and researchers who have already and recently used a microscope to acquire images of fluorescent samples, to improve their microscopy skills.
The course is NOT aimed at training people to use the LCI facility microscopes.ย The focus is instead on providing the students with enough theoretical and practical knowledge about their OWN sample and their OWN microscope, to enable them to:

1) prepare their sample and formulate their scientific question in a way that is suitable for data extraction from fluorescence images

2) properly use the hardware available in their lab/facility and 3) fully understand each parameter they need to set in the software in their lab/facility.

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

Please help us spread the word.ย ๐Ÿ˜ƒ

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 Wรคhlby 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 registerย here.

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