Sylvie Le Guyader

Karolinska Institutet | KI · Department of Biosciences and Nutrition

1-      This Wednesday (16.03), you are welcome to join 2 events organized at the LCI facility:

a.       09:15-09:45: A short seminar on Optogenetics by Alexandr Pospech from Andor.

Optogenetics is a combination of genetics and optics to modulate ion flow in targeted cells within a tissue or live animals using a light beam on a microscope. Applications target neurons but also cardiomyocytes or stem cells.

b.      10:30-11:30: A seminar on the Imaris image analysis software

The seminar will present the power of the Imaris software to extract quantitative information from microscopy images (2D, volumes, timelapses, multi colour…) as well as represent the data using multi dimensional plots.

Both seminars will be held in the Red seminar room, at Bionut (Hälsovägen 7, lift E, 6th floor, the door will be open 5 min before the start of each seminar).

2-      As I realized that many of you working with animals do not know about the Preclinical Imaging Facility here at KI Huddinge, I want to advertise for them: Using their equipment and expertise, you can image small live animals in an non-invasive way so that you can follow e.g. cancer progression over weeks. Contact Ying (ying.zhao.1(at)ki.se) for further information.

3-      We still have a few spots available for the Live Cell Imaging microscopy course (KI courses #2870 and #2871) starting on the 7th of April.

If you want to join the course, email me. Otherwise you are anyway welcome to come and listen to the lectures without registration. The full schedule is now available online on our website and will be kept up to date in case we must change something but it is pretty final. On our website you can also find instructions on how to find us. The Green seminar room, where the lectures will be held, will be clearly indicated within our Novum building. Please spread the word (poster attached). Tack :)       

4-      Embedded into our course, we will have a light sheet microscopy seminar, workshop and demo by LaVision Biotech. You can join the seminar during the course on the 15.04, 11:40-12:10 (see point 3) and/or email me if you want to try and image your own sample on the LVBT system as it will stay at our facility until the 21st of April.

5-      Antibodypedia is a useful database of human antibodies that have been validated by users. If you decide to validate your favorite antibody and you publish the results on their site, they apparently pay back part of the cost of antibody production. At least this is what I have heard but I could not find anything on their webpage (see link) about this. :(

6-      Bumped into the Switch technique (see link) to label a tissue with up to 20 antibodies in multiplexing imaging :)

1-      We still have a few spots available for our advanced microscopy courses (#2870 and #2871) in April 2016.

The LCI facility runs 2 microscopy courses in parallel:

-          KI course #2870 with theory and workshops, some using your own sample so that you get all the tips and tricks that are relevant to your project (4.5 points).

-          KI course #2871 with theory only (3 points).

All details are found on our website.

2-      Many people talk about Phenol Red when mentioning cell medium autofluorescence. Actually, most medium autofluorescence is green, not red, and it is caused by flavins and vitamin B12 in the medium. If you are imaging dim green fluorescence, you can try a medium with less flavins. For example the second link sends you to a paper that talks about the advantages of using F12 (10 times less riboflavin than DMEM). Another alternatives are Fluorobrite, LiveLight or something from Evrogen (see links).

3-      The 6th link points to a website that might give some useful tips to those of you imaging RNA FISH.

4-      Hans Blom organizes one of his great workshops. This time it is about Live 3D imaging using multifocus microscopy.

5-      If you would like to stain your favorite tissue with zillions of antibodies and have a hard time figuring out a protocol, you can turn to the expertise of the SciLife Fluorescence Tissue Profiling facility headed by Jan Mulder. see the link.

6-      If you would like to read more such tips and tricks, register your email to receive the RSS feed of the excellent microscopy blog by Kurt Thorn (see link)

1-      The Live Cell Imaging facility microscopy course got a twin brother! :)

From now on, the LCI facility will run 2 microscopy courses in parallel:

-          KI course #2871 with lectures only (3 points, see first link)

-          KI course #2870 with the same lectures but additional exercises and workshops using your own sample (4.5 points, see second link).

The courses will run in April 2016. you can find the program, list of lecturer... on our website (or see third link). Apply via KI or email me.

Please spread the word. Tack :)       

2-      The Biovis imaging facility in Uppsala organizes super soon 1 imaging course (30.09-09.10) and 1 image processing course (24-27.11). Contact them directly to check if there are spaces left.

3-      On 26.10/2015, in Flemingsberg we will celebrate the inauguration of the Jonasson center, a center dedicated to funding medical and research imaging equipment (PET, MRI, confocals…) in Huddinge. You are welcome to join the inauguration and the symposium.

4-      Some of you might know about this but just in case, the fourth link leads to an article showing how to remove autofluorescence in some tissue using Sudan Black. We have done a little comparison at our facility and it worked very nicely for thin paraffin sections of Salamander heart.

5-      Pontus Nordenfelt from Lund University will give a talk at KI Bionut (26.10 at 10:45, red seminar room. Ask me if you need directions). Pontus recently came back from Harvard where he worked with Tim springer and Clare Waterman. His focus is on cell migration and integrins in focal adhesions. He images in TIRF using polarized light to detect the orientation of the labelled molecule then runs fancy automated image analysis.

1-      It will soon be time to apply to the Live Cell Imaging facility course: ‘Microscopy: how to improve your imaging skills? Theory and practical tips from sample preparation to image analysis’.

The course will be announced in the fall on the KI course catalog (course #2820) and will run in the spring 2016. You can find the program of the 2015 course on the KI course catalog. Please spread the word.

2-      Dirk Pacholsky, the manager of the Biovis imaging facility at Uppsala University, has created a user group for people who perform optical clearing of their sample before imaging. Clearing is a group of techniques that make thick samples more transparent to help imaging with light microscopy. If you would like to join the Biovis Clearing User Group and exchange clearing tips, please contact Pacho directly (Dirk Pacholsky <dirk.pacholsky@scilifelab.uu.se>).

3-      Carolina Wählby is a Professor in Image Analysis at Uppsala University/SciLife. She will run one of her excellent image analysis courses in the autumn this year. If you are interested, please check this link and apply directly.

4-      If you have back/shoulder/neck problems when working at your computer (who doesn’t?), I would like to recommend a little freeware that I bumped into recently: Workrave.

It takes very little PC power and it forces you to take breaks by locking your screen, for example 30 sec every 30 min or whatever you set. You get a reminder to stretch and get a chance to follow some exercises for the back, the eyes… Hope there is no virus in it but I (and Symantec) have noticed nothing abnormal so far.

5-      For those who run migration assays in transwells, I have heard of the Fluoroblok transwells that prevent fluorescence from cells at the top of the well. This way there is no need to scrape before imaging.

1-      I would like to announce a workshop organized at the LCI unit on the 8th of June at 13:00 and presenting the CellDirector chamber sold by a GradienTech. This is a perfusion chamber that allows formation and imaging of controlled 2D and 3D gradients in cell culture. This chamber can be mounted on a microscope and the reaction of the live cells to the gradient imaged over time. Check the link to watch demo videos.

2-      There is also a super resolution symposium on the 2nd of June at SciLife Solna. Here you can find the program. Please contact Hans Blom directly if you have any question.

This is the newsletter of the Live Cell Imaging core facility at the Karolinska Institute in Sweden and it is all about light microscopy.

Each newsletter will contain:

Please feel free to send me some information about your own light microscopy events so that I can add it here. Remember to inform Swedish Bioimaging as well so that they can put it up on their website.

If you want to be automatically updated when I post something, please follow my Research Gate page. Note that pressing the Follow button near one of the questions will not keep you updated as each newsletter will be posted as a new question for clarity sake.

Feel free to spread the word if you think this could help others! :)

Sylvie