Monica Ballarino
University of Rome, La Sapienza
Italy
EMBO Practical Course
Over the past decades, it became evident that the specific amount and localisation of an RNA transcript are important to understand the biological function of its (non-) coding gene. Whereby state-of-the-art in situ hybridisation (ISH) technologies are key to visualise these RNAs in a given tissue or cell.
This course will teach participants to apply three powerful RNA visualisation techniques, all in histological tissue samples: conventional in situ hybridisation (ISH), hybridisation chain reaction ISH (HCR-ISH), and the cutting edge single molecule fluorescent ISH (smFISH). In addition, essential tissues embedding protocols adapted to facilitate visual interrogation of RNA transcripts will also be covered.
Benefit from our excellent trainer to participant ratio for personalised training in addition to world class research facilities and networking opportunities with leading experts in the field.
This course is targeted to early stage scientists (PhD and postdocs) across the field of molecular biology with an interest in application of these techniques for their research.
This hybrid course will begin on 6 October 2023, with a joint onboarding session for on-site and virtual participants – where access to pre-course study materials, exercises and pre-recorded lectures will be provided through an introduction to our collaboration platform EMBL eCampus. This will prepare the participants for the course start on 22 October 2023.
Full-time attendance is required for both virtual and on-site components of this course.
Information on the live stream and access to the virtual course platform will be provided shortly before the start of the event and will be available for 2 weeks after the course.
All lectures will be delivered on site and live-streamed to all virtual participants via Zoom. Please see the preliminary programme for details on all lectures.
These are the practicals for the course, all of the content will be delivered on-site in the training laboratories and the microscopy rooms at EMBL. The practicals will not be live-streamed to virtual participants, please see below for the virtual practical information which will take place in tandem to the on-site practicals:
The content of the virtual practicals will be delivered in the form of interactive protocols as well as video tutorials recorded by the trainers. The following content will be available for virtual participants during the above on-site practical sessions. Please see the preliminary programme for when these will take place:
The interactive protocol will cover NcRNA or microRNA in situ hybridisation on paraffin sections of mouse tissues.
We will also include Q&A sessions with trainers to give participants the opportunity to ask questions about the procedures of the material given.
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Please see EMBL’s COVID-19 safety policy for testing and other requirements to attend this on-site course.
University of Rome, La Sapienza
Italy
University of Trento
Italy
University of Nice, Institut de Biologie Valrose (iBV)
France
EMBL Heidelberg
Germany
University of Rome, La Sapienza
Italy
Friedrich Miescher Institute for Biomedical Research
Switzerland
Max Delbrück Center for Molecular Medicine
Germany
Center for Life Nano- & Neuro-Science of Istituto Italiano di Tecnologia (IIT)
Italy
Münster University Hospital
Germany
Institute of Microbiology Heinrich Heine University
Germany
Turbine.ai
Hungary
Max Planck Institute of Biochemistry
Germany
Princeton University
USA
EMBL Heidelberg
Germany
Max Planck Institute of Biochemistry
Germany
EMBL Rome
Italy
University of Rome, La Sapienza
Italy
University of Cambridge
UK
University of Rome, La Sapienza
Italy
Turbine.ai
Hungary
EMBL Rome
Italy
Course and Conference Officer
EMBL Heidelberg
Germany
Training Lab Technician
EMBL Heidelberg
Germany
Please note – This programme is preliminary at the moment and is subject to change.
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Time (Europe/Berlin) | Programme |
Milestones of the day | Milestones of the day: -How to use eCampus -Explain milestones -How to follow the workflow -How to use the protocol -How to use the resources -VMs -Watch pre-recorded lectures by Sunday 22nd October 2023 – the start of the course (eCampus, lectures 1, 2, 3 ) |
13:00 – 13:20 | Welcome – virtual onboarding |
13:20 – 13:40 | Practical parts – Online Protocols |
13:40 – 14:40 | Ice breaker |
14:40 – 15:00 | Wrap up |
Time (Europe/Berlin) | Onsite Programme | Virtual Programme |
Milestones of the day | Milestones of the day: None | |
13:45 – 14:00 | Bus from ISG hotel to EMBL | Log on and check connection |
14:00 – 14:30 | Welcome and house notes and course overview | Welcome and house notes and course overview |
14:30 – 16:30 | Ice breaking activity – Flash talks and activity in groups with concurrent coffee break | Ice breaking activity – Flash talks and activity in groups |
16:30 – 17:15 | Keynote Lecture 1: Applying RNA imaging methodologies to tackle ncRNAs in brain Marie-Laure Baudet – University of Trento, Italy | Keynote Lecture 1: Applying RNA imaging methodologies to tackle ncRNAs in brain Marie-Laure Baudet – University of Trento, Italy |
17:15 – 18:00 | Keynote Lecture 2: Transcriptional profiling of spermatogenesis in human and issues in infertility. Sara Di Persio – Münster University Hospital, Germany | Keynote Lecture 2: Transcriptional profiling of spermatogenesis in human and issues in infertility. Sara Di Persio – Münster University Hospital, Germany |
18: 00 – 18:45 | Comfort break | Comfort break |
18:00 – 21:00 | Moderated discussion rounds with concurrent dinner in the canteen | Moderated discussion rounds – The onsite participants will also eat during the time, please feel free to do this too. |
18:45 – 19:30 | Discussion to Prerecorded LECTURE 1 – Topic: Histology overview and applications Moderated by: Emerald Perlas – EMBL Rome, Italy | Discussion to Prerecorded LECTURE 1 – Topic: Histology overview and applications Moderated by: Emerald Perlas – EMBL Rome, Italy |
19:30 – 20:15 | Discussion to Prerecorded LECTURE 2 – Topic: RNA detection methods Moderated by: Imre Gaspar – Turbine.ai, Hungary | Discussion to Prerecorded LECTURE 2 – Topic: RNA detection methods Moderated by: Imre Gaspar – Turbine.ai, Hungary |
20:15 – 21:00 | Discussion to Prerecorded LECTURE 3 – Topic:ncRNA Charme in cardiac and skeletal muscle Moderated by: Monica Ballarino, University of Rome, La Sapienza, Italy | Discussion to Prerecorded LECTURE 3 – Topic:ncRNA Charme in cardiac and skeletal muscle Moderated by: Monica Ballarino, University of Rome, La Sapienza, Italy |
21:00 – 21:15 | Wrap up | Wrap up |
21:15 | Departure to Hotel |
Time (Europe/Berlin) | Onsite Programme | Onsite location | Virtual Programme |
Milestones of the day | Milestones of the day: Practical 1: Introduction and Preparation for practical e.g. buffer prep Practical 1: Protocol steps 1-5 Practical 2: Protocol and Video: Dye conjugation of amino-11-ddUTP (12 min) | ||
08:30 – 09:00 | Arrival and coffee | ||
09:00 – 09:10 | Overview of the day | Flex labs | Overview of the day |
09:10 – 09:30 | Lab safety instructions | Flex labs | Lab safety instructions |
9:30 – 10:45 | Interactive lecture/demo: Specimen section preparation Emerald Perlas – EMBL Rome, Italy | Training Lab A & B | LIVESTREAM PRACTICAL: Interactive lecture/demo: Specimen section preparation Emerald Perlas – EMBL Rome, Italy |
10:45 – 11:00 | Coffee break | Flex labs | Emerald continues and gives intro to conventional ISH for virtual participants, Imre gives an overview too |
11:00 – 11:45 | Keynote Lecture 7: Neuronal sorting of RNA. Marina Chekulaeva – Max Delbrück Center for Molecular Medicine, Germany | Flex labs | Keynote Lecture 7: Neuronal sorting of RNA. Marina Chekulaeva – Max Delbrück Center for Molecular Medicine, Germany |
11:45 – 12:30 | Keynote Lecture 4: Interplay between genetic and environmental factors in shaping body plans. Aissam Ikmi – EMBL Heidelberg, Germany | Flex labs | Keynote Lecture 4: Interplay between genetic and environmental factors in shaping body plans. Aissam Ikmi – EMBL Heidelberg, Germany |
12:30 – 13:30 | Lunch break | EMBL Canteen | |
13:30 – 14:15 | Practical session 1: Introduction and theory of conventional ISH. Protocol Overview. Emerald Perlas assisted by Chiara Capponi, Sara Di Persio, and Tiziana Santini | Training Lab A & B | |
14:15 – 14:45 | Practical session 2: Probe preparation (ddUTP + dye conjugation) Imre Gaspar assisted by 3 helpers | Training Lab A & B | |
14:45 – 16:45 | Practical session 1: Conventional ISH; (section pre-treatment, Proteinase K digest, acetylation, hybridization set-up). Emerald Perlas assisted by Chiara Capponi, Sara Di Persio, and Tiziana Santini | Training Lab A & B | |
16:45 – 17:00 | Coffee break | Flex labs | |
17:00 – 17:45 | Keynote Lecture 5: Phase separation in RNA localization. Mainak Bose – EMBL Heidelberg, Germany | Flex labs | Keynote Lecture 5: Phase separation in RNA localization. Mainak Bose – EMBL Heidelberg, Germany |
17:45 – 18:00 | Comfort break | ||
18:00 – 19:30 | Dinner | EMBL Canteen | |
19:30 – 21:30 | Student presentations: 10 student presentations (each: 5 min presentation plus 6 min discussion) | Flex labs | Student presentations: 10 student presentations (each: 5 min presentation plus 6 min discussion) |
21:40 | Departure to Hotel | ATC Entrance |
Time (Europe/Berlin) | Onsite Programme | Virtual Programme |
Milestones of the day | Milestones of the day: Practical 3a and 3b: -Video: smFISH probe design (part1, part 2, part3) (36 min) -Video: HCR design (part1,part2) (43 min) -OPTIONAL: Preparation of buffers for lab -Protocol steps ‘probe labelling for smFISH’ -Video: probe labelling smFsFISH (part 1, part 2, part 3) (22 min) -Video: sm FISH and HCR-ISH specimen pre-work and hybridisation set-up (8 min) Practical 1: Protocol steps 6-end | |
08:30 – 09:00 | Arrival and coffee | |
09:00 – 09:10 | Overview of the day | |
09:10 – 10:15 | Practical session 1: (conventional ISH) post-hybridisation washes Emerald Perlas assisted by Chiara Capponi, Sara Di Persio, and Tiziana Santini | |
10:15 – 11:00 | Keynote Lecture 3: RNP granule dynamics Florence Besse – University of Nice, Institut de Biologie Valrose (iBV), France | Keynote Lecture 3: RNP granule dynamics Florence Besse – University of Nice, Institut de Biologie Valrose (iBV), France |
11:00 – 11:15 | Coffee break | |
11:15 – 12:00 | Practical session 1: Conventional ISH: staining set-up Emerald Perlas assisted by Chiara Capponi, Sara Di Persio, and Tiziana Santini | |
12:00 – 13:00 | Lunch at EMBL canteen | |
13:00 – 13:30 | Practical session 2: Theory and demo of probe design Imre Gaspar assisted by 3 helpers | LIVESTREAM PRACTICAL – Practical session 2: Theory and demo of probe design Imre Gaspar assisted by 3 helpers |
13:30 – 14:30 | Practical 3a and 3b: Introduction and theory smFISH and HCR Monica Ballarino, Imre Gaspar, Eleonora D’Ambra, Tiziana Santini | LIVESTREAM PRACTICAL – Practical 3a and 3b: Introduction and theory smFISH and HCR Monica Ballarino, Imre Gaspar, Eleonora D’Ambra, Tiziana Santini |
14:30 – 15:00 | Coffee break | |
15:00 – 16:00 | Practical session 1: Conventional ISH: washing and mounting of specimen Emerald Perlas assisted by Chiara Capponi, Sara Di Persio, and Tiziana Santini | |
16:00 – 16:30 | Practical 3a and 3b: sm FISH and HCR-ISH specimen pre-work and hybridisation set-up Monica Ballarino, Imre Gaspar, Eleonora D’Ambra, Tiziana Santini | |
16:30 – 17:00 | Practical 3a: sm FISH: Probe labeling set-up Monica Ballarino, Imre Gaspar, Eleonora D’Ambra, Tiziana Santini | ON incubation sm labeling! |
17:00 – 18:00 | Practical 3a and 3b: continued: HCR-ISH specimen pre-work and hybridisation set-up Monica Ballarino, Imre Gaspar, Eleonora D’Ambra, Tiziana Santini | |
18:00 – 19:00 | Dinner at EMBL | |
19:00 – 21:10 | 11 student presentation (each: 5 min presentation plus 6 min discussion) | 11 student presentation (each: 5 min presentation plus 6 min discussion) |
21:15 | Departure to Hotel |
Time (Europe/Berlin) | Onsite Programme | Virtual Programme |
Milestones | Milestones of the day: Practical 3a and 3b: -Protocol parts: ‘smFISH in myotubes and ‘HCR’ -Video: smFISH and HCR generics (part 1, part2) (23 min) -Video: Mounting? | |
08:30 – 09:00 | Arrival and coffee | |
09:00 – 09:10 | Overview of the day | |
09:10 – 11:30 | Practical 3a and 3b: smFISH: Probe purifications and measurements HCR-ISH: washes smFISH pre-hybridisation set-up for incubation HCR-ISH: Amplification set-up for reaction smFISH hybridisation Monica Ballarino, Imre Gaspar, Eleonora D’Ambra, Tiziana Santini With concurrent coffee break | |
11:30 – 12:15 | Keynote Lecture 8: Adaptable P body physical states differentially regulate bicoid mRNA storage during early Drosophila development Tim Weil – University of Cambridge, UK | Keynote Lecture 8: Adaptable P body physical states differentially regulate bicoid mRNA storage during early Drosophila development Tim Weil – University of Cambridge, UK |
12:15 – 13:15 | Lunch break at EMBL canteen | |
13:15 – 15:45 | Practical session 1: Conventional ISH imaging Emerald Perlas assisted by Chiara Capponi, Sara Di Persio, and Tiziana Santini With concurrent coffee break | LIVESTREAM Practical with Emerald and Trainers |
15:45 – 16:30 | Keynote Lecture 9: Membrane associated RNA transport Michael Feldbruegge – Institute of Microbiology Heinrich Heine University, Germany | Keynote Lecture 9: Membrane associated RNA transport Michael Feldbruegge – Institute of Microbiology Heinrich Heine University, Germany |
16:30 – 18:00 | Practical 3a and 3b: smFISH & HCR-ISH: washes and specimen mounting Monica Ballarino, Imre Gaspar, Eleonora D’Ambra, Tiziana Santini With concurrent coffee break | |
18:00 – 19:00 | Dinner at EMBL | |
19:00 – 20:30 | DISCUSSION ROUND (responsible conduct of research) Experimental design Standards and controls in the ISH field | DISCUSSION ROUND (responsible conduct of research) Experimental design Standards and controls in the ISH field |
20:40 | Departure to Hotel |
Time (Europe/Berlin) | Onsite Programme | Virtual Programme |
Milestones of the day | Milestones of the day: Prepare for discussion rounds with Emerald, Monica and co. | |
08:30 – 09:00 | Arrival and coffee | |
09:00 – 09:10 | Overview of the day | |
09:10 – 10:30 | Practical 4: Imaging in groups ( smFISH and HCR-ISH) Monica Ballarino, Imre Gaspar, Eleonora D’Ambra, Tiziana Santini | |
10:30 – 10:45 | Coffee break and group photo | Group photo – Please take a screen shot of yourselves on Zoom, we will add this to the group photo. |
10:45 – 12:15 | Practical 4: Imaging in groups ( smFISH and HCR-ISH) Monica Ballarino, Imre Gaspar, Eleonora D’Ambra, Tiziana Santini | Meet the speaker for the virtual participants |
12:15 – 13:15 | Lunch at EMBL canteen, table set-up based on next career steps | |
13:15 – 14:00 | Keynote Lecture 10: RNP assembly for transport and translational control Elizabeth Gavis – Princeton University, USA | Keynote Lecture 10: RNP assembly for transport and translational control Elizabeth Gavis – Princeton University, USA |
14:00 – 14:45 | Keynote Lecture 6: Dynamics of localized translation of mRNA Jeffrey Chao – Friedrich Miescher Institute for Biomedical Research, Switzerland | Keynote Lecture 6: Dynamics of localized translation of mRNA Jeffrey Chao – Friedrich Miescher Institute for Biomedical Research, Switzerland |
14:45 – 16:15 | Meet the speakers for the on-site participants | LIVESTREAM – Practical 4: Imaging ( smFISH and HCR-ISH) with Imre Gaspar |
16:15 – 16:30 | Coffee break | Coffee break |
16:30 – 17:15 | Keynote Lecture 11: PAINT superresolution imaging of nucleic acids Ralf Jungmann – Max Planck Institute of Biochemistry, Germany | Keynote Lecture 11: PAINT superresolution imaging of nucleic acids Ralf Jungmann – Max Planck Institute of Biochemistry, Germany |
17:30 – 18:00 | Bus to Heidelberg | |
18:00 – 19:30 | Heidelberg Guided City tour | |
19:30 -22:30 | Dinner at Kulturbrauerei in downtown Heidelberg |
Time (Europe/Berlin) | Onsite Programme | Virtual Programme |
Milestones of the day | Milestones of the day: None | |
08:30 – 09:00 | ||
09:00 – 10:00 | Discussion on Practical 4: Summary of Image taking, including best practices and standards Monica Ballarino, Imre Gaspar, Eleonora D’Ambra, Tiziana Santini | |
10:00 – 10:30 | Coffee break | |
10:30 – 11:50 | Practical session 5: Basics ISH image analysis Imre Gaspar assisted by 3 helpers | LIVESTREAM – Practical 5: Basics ISH image analysis with Imre Gaspar |
11:50 – 13:20 | Practical session 5: ISH image analysis (incl. standards in the field) Imre Gaspar assisted by 3 helpers | LIVESTREAM – Practical 5: ISH image analysis with Imre Gaspar |
13:20 – 14:20 | Lunch at EMBL canteen | |
14:20 – 15:20 | Group work: Participants preparing group presentations supported by trainers with concurrent coffee break | Virtual participants welcome to join, but no virtual presentation possible |
15:20 – 16:20 | Discussion: Group Data Presentations incl. Discussions with concurrent coffee break | Discussion: Group Data Presentations incl. Discussions |
16:20 – 17:20 | Wrap up | Wrap up |
17:45 | Bus to Heidelberg train station and city center |
Time (Europe/Berlin) | Onsite Programme | Virtual Programme |
13:00-15:00 | Postcourse day: Virtual session – Open discussion round on questions to practicals (first questions will be collected via EMBL eCampus) All main-trainers | Postcourse day: Virtual session – Open discussion round on questions to practicals (first questions will be collected via EMBL eCampus) All main-trainers |
The course is limited to 16 onsite participants and 5 virtual. For selection purposes, please note that your application will not be considered without a letter of motivation.
On-site registration fees include admission, course materials, meals and coffee breaks. This EMBO course includes accommodation and transportation to and from the ISG Hotel to the venue.
Virtual registration fees include access to live-streamed course content (lectures, Q&A sessions with speakers and trainers, discussion rounds) as well as virtual learning activities in the learning platform.
On-site Academia | €500 |
On-site PhD Student | €500 |
On-site Industry | €1200 |
Virtual Academia | €190 |
Virtual PhD Student | €190 |
Virtual Industry | €250 |
A letter to support your visa application will be issued, on request, once payment of the registration fee is confirmed. We recommend that you book your visa appointment as soon as possible, to avoid any delay with your visa application.
The registration fee should be paid only after acceptance to the course. The results will be announced approximately 2-3 weeks after the application deadline.
After you have logged in and successfully registered, you will receive an email asking you to submit your motivation letter. Click on the link provided and enter your motivation letter in the text box provided. Alternatively, you can submit your motivation letter by clicking on the link on the confirmation page directly after registering.
Instructions
Please note:
For detailed instructions, please watch our video on how to submit a course motivation letter.
For further information about registration and motivation letter submission please refer to the FAQ page.
Limited financial assistance is provided by the EMBL Advanced Training Centre Corporate Partnership Programme and EMBO in the form of both registration fee waivers and travel grants. Availability is limited to participants attending on-site EMBL Courses and virtual or on-site EMBO Practical Courses in Heidelberg, and will be indicated during the motivation letter submission process.
Your place in the course is only confirmed by paying the registration fee, which is mandatory even when receiving a fee waiver.
The fee waiver will cover the registration sum that you have paid to attend the course.
The travel grant will cover the cost of travel (airfare, train, bus, taxi, accommodation, visa, and/or registration fees*) and is provided up to specified caps which are normally as follows:
– up to €400 for participants travelling to an EMBL Course within Europe.
– up to €1000 for participants travelling to an EMBL Course from outside Europe.
– up to €500 for any participant travelling to an EMBO Practical Course.
– up to €1000 for any participant working in Chile, India, Singapore or Taiwan travelling to an EMBO Practical Course.
– up to €700 for any participant working in Croatia, Czech Republic, Estonia, Greece, Hungary, Italy, Lithuania, Luxembourg, Poland, Slovenia, and Turkey travelling to an EMBO Practical Course.
*Registration fees are only covered for EMBO Practical Courses
The organisers may reduce the grant cap to accommodate more participants. Recipients will be notified of their travel cap amount when they are informed of the outcome of their application. Original receipts must be provided with your signature for all costs incurred within two months of completion of travel. Scanned copies cannot be accepted.
For EMBO Practical Course participants with children, there is the possibility to apply for a childcare grant to offset childcare costs incurred by participants or speakers when participating at a course. Eligible costs include fees for a babysitter or child-care facility, travel costs for a caregiver, or travel costs for taking the child to the meeting etc. Please note that priority will be given to early-stage researchers. A maximum amount of €500 can be awarded per participant applying for an EMBO childcare grant.
Applies to selected courses only. Availability will be indicated during the motivation letter submission process.
This grant covers costs related to your attendance at the course (registration, travel and accommodation costs). The grant is restricted to PhD students and postdocs who conduct basic biomedical research.
Whether you are eligible to apply for a travel grant, depends on when you received your university entrance qualification (e.g. Abitur, A-Levels, High School Diploma, Final State Examination):
– for PhD and MD students, as well as graduates, the university entrance qualification must not have been obtained more than 11 years ago at the time of the envisaged course
– for postdocs, the university entrance qualification must not have been obtained more than 13 years ago at the time of the envisaged course
You may apply for financial assistance when submitting your motivation letter for courses. In your application, you will be asked to answer questions regarding why your lab cannot fund your attendance and how your attendance will make a difference to your career. Application for financial support will not affect the outcome of your registration application.
For the Boehringer Ingelheim Fonds Travel Grant, there is a pre-application question during the motivation letter submission process, and if selected you will be requested to complete a standard form and documentation consisting of your travel expense estimation.
The scientific organisers will select the recipients of all financial assistance during the motivation letter selection process. Results will be announced approximately 6-8 weeks before the event start date, however for some events this may be delayed. Selection results do not impact your admission to the course. Selection is based on scientific merit, your current work or study location, the reasons for needing financial support, and the impact this event will have on your career.
Costs will be reimbursed after the meeting only once a reimbursement form and original receipts (from travel costs) have been received.
See our list of external funding opportunities and for further information about financial assistance please refer to the FAQ page.
Accommodation has been pre-booked for you at the ISG Hotel for the default dates of Sunday 22 October to Friday 27 October 2023 (5 nights).
Please note that the course registration fee includes coverage of accommodation costs in twin rooms, including breakfasts, for your stay at the ISG. In case you would like to have a single room, please let us know and please note that you will be required to pay an additional cost of 24€ per night. This should be paid by you directly to the ISG during your stay. Any additional nights need to be booked and paid for by you.
We recommend that all course participants stay at the ISG Hotel which is the dedicated hotel for the course.
Shuttle buses will go from the ISG Hotel to EMBL and back, mornings and evenings. A bus schedule and location of the bus stops will be made available prior to the meeting.
Address: EMBL Heidelberg, Meyerhofstraße 1, 69117 Heidelberg, Germany
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A variety of activities in Heidelberg can be found on the website of Heidelberg Marketing.
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Event software
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Date: 22 - 27 Oct 2023
Location: EMBL Heidelberg and Virtual
Venue: EMBL Advanced Training Centre
Deadline(s):
Application: Closed
Application: Closed
Organisers:
Contact: Charlotte Courtney