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!

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.

Cool stuff one can do with the LCI Primo

At the LCI core facility, you can use a machine called Primo to micropattern/print proteins at the bottom of a microscopy dish and enable real time imaging of cell-protein interactions.

Now we learnt that we can even micropattern lipids and antibodies on microscopy dishes to image how cells interact with these molecules! This can be done with any pattern and even in multiwell plates! 🙂

Lipid micropatterning: Here is a cool paper showing how filopodia interact with sourrounding lipids and proteins. They image with Structured Illumination and TIRF, techniques that are also on offer at our facility. 🙂

Antibody micropatterning: A new way to analyse extracellular vesicles with multiplexed detection of proteins and RNAs at single EV resolution.

Light-seq: Multiplexed, non-destructive spatial transcriptomics of tissues sections using light

Light-Seq is a new pretty cool technique for highly multiplexed sequencing of RNA in tissue sections using light. This technique is highly sensitive, highly spatially resolved and because it does not destroy the tissue, it can be combined with protein labelling (genetic or by immunolabelling).

On one of our single-point confocal/spinning disk/widefield system at the LCI facility, we have a device called Primo (DMD + UV laser) which can be used to run this technique! 🙂

Let us know if you would like to set up LightSeq at the LCI core facility!

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! 🙂

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.

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! 😊

Hope you enjoy the LCI facility microscopy course 2022!

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:

Crest V3 spinning disk confocal demo

Tomorrow (17 sept) we will enjoy a seminar and a live demo about the Crest V3 spinning disk confocal which is being set up at our facility as I write! 😀

Very cool confocal!

  • enormous field of view (32 mm diameter)
  • fully confocal
  • can image at 100 frames per sec
  • spits out Nyquist resolution with the 60x objective!

You can come to the seminar (at 10 in the Gene seminar room at the LCI facility) or listen to it remotely (see here how to follow the LCI webinars).

You can even book a private demo to image your own samples.

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!!

Follow us! :-)

Type your email address below to subscribe to our blog!