Scientific and Financial Report

Transylvanian Experimental Neuroscience

Summer School (TENSS 2015)
www.tenss.ro
June 1-19, 2015, Pike Lake, Romania

Raul C. Murean

Florin Albeanu

coneural

Center for Cognitive and Neural Studies

Romanian Institute of Science and Technology

Adam Kampff

- June 24, 2015 -

Table of contents
1. Testimonials about TENSS 2015 ................................................................................................... 1
2. Scientific content of the event ..................................................................................................... 2
2.1. Objectives .............................................................................................................................. 2
2.2. Selection of students............................................................................................................. 2
2.3. General structure of the program......................................................................................... 5
2.4. Lab sessions ........................................................................................................................... 6
2.5. Lectures ............................................................................................................................... 10
2.6. Some pictures from TENSS 2015 ......................................................................................... 13
2.7. Contribution of TENSS to Neuroscience and future directions .......................................... 54
3. Information regarding the organizing of the event ................................................................... 55
4. Financial report .......................................................................................................................... 57
5. Final program of the event ........................................................................................................ 58

1. Testimonials about TENSS 2015

TENSS fully met my expectations. And more. I have done other schools in the past. But this is
the first one that gave me the tools to do things myself. The technical and scientific level was
excellent and I think I have acquired more understanding than what I can express at this
point. The personal and relaxed vibe of the school is fantastic and you should always keep it. I
felt really comfortable and happy. Enjoyed both the scientific and non-scientific aspects of
this very social school.
TENSS 2015 student A
The mix of lectures, labs & social activities was very well balanced in order to achieve the
goals. I especially liked that there was enough time for discussions & interactions in between
the lectures and labs. TENSS was 19 days of intense work & a lot of fun I feel very grateful
for getting a lot of knowledge, insights & skills! I actually did not expect it to be as awesome
as it turned out to be. In addition to great lectures & labs, the atmosphere was inspiring,
relaxed and fun. It requires a lot of work & passion to set up such a great course and I
appreciate it. Thanks a lot!
TENSS 2015 student B
Everything that I learned in this course will have a great impact on my research given my
situation starting at experimental work, this school was absolutely perfect for me. About
TENSS, I liked most the environment of freedom, the feeling that we can do whatever we
want, the fact that this freedom leads to being very creative and having amazing new ideas,
the flexibility of the organizers that allowed for this.
TENSS 2015 student C
I learned all the techniques I wanted and realized that even if Im not familiar with
something, it can be learned. I really enjoyed the informality and the flexibility. TENSS helped
me to gain back my enthusiasm for science!
TENSS 2015 student D
1

2. Scientific content of the event

2.1. Objectives
The Transylvanian Experimental Neuroscience Summer School (TENSS) was born in 2010,
from an idea that Florin Albeanu shared to Raul Murean during a conference in Romania.
The first edition of TENSS took place in 2012, on the shores of Pike Lake, in the picturesque
province of Transylvania. Starting with its third edition in 2014, TENSS has welcomed a third
permanent organizer, Adam Kampff, from University College London, UK.
The main objective of TENSS is to provide participants top-level, hands-on training in
experimental techniques used in neuroscience, from optical all the way to electrophysiological methods and behavioral setups. To the best of our knowledge the opportunity
for such training is scarce and is covered only in a few places in the world, such as Cold Spring
Harbor and Woods Hole, in the USA or Plymouth in UK, for example. A second objective of
TENSS is to attract a large number of top-level scientists from all over the world, who can
provide theoretical lectures to participants and can interact with participants, among
themselves, and possibly with local scientists based in Romania, giving rise to scientific
debates of the highest quality. Finally, TENSS aims to bring more world-wide visibility to
neuroscientific research in both Europe and in Romania.

2.2. Selection of students

TENSS 2015 has received 150 applications with students originating from 42 countries,
distributed as follows (as a function of the citizenship of students):
Argentina: 4 applications
Armenia: 3 application
Austria: 3 applications
Brazil: 6 applications
Chile: 3 applications
China: 2 applications
Columbia: 3 applications
Croatia: 1 application
2

 Egypt: 1 application
Finland: 2 applications
France: 1 application
Georgia: 1 application
Germany: 12 applications
Greece: 1 application
Hungary: 5 applications
India: 16 applications
Iran: 6 applications
Ireland: 2 applications
Israel: 3 applications
Italy: 10 applications
Kenya: 1 application
Korea: 1 application
Mexico: 1 application
Morocco: 1 applications
Nigeria: 4 applications
Norway: 2 applications
Pakistan: 4 applications
Poland: 4 applications
Portugal: 2 applications
Romania: 5 applications
Russia: 6 applications
Singapore: 1 application
Slovak Republic: 1 application
Spain: 3 applications
Sweden: 2 applications
Switzerland: 2 applications
Taiwan: 1 application
Turkey: 1 application
United Kingdom: 7 applications
Uruguay: 1 application
3

 United States of America: 13 applications

The selection process has been performed by 15 evaluators (organizers and TAs) distributed
in several labs around the world. We initially created 5 groups of 3 evaluators for screening
the applications and selecting the top 50-60 ones. Each group selected 10-12 applications that
made it in the second round. The second round thus contained a total of 56 applications that
were evaluated by all 15 evaluators, to level out individual preferences and eliminate as much
as possible subjective bias. We selected initially 13 top students by mixing those from wellestablished labs with promising and talented students from less favored countries, such as
places in Eastern Europe and Asia. From the initial selection, one student from MRC
Laboratory of Molecular Biology in UK could not attend because of personal reasons
and was replaced by the next student on the waiting list. Also, one student from NCBS
in Bangalore India could not attend the course because he did not receive a
Romanian visa in time. Therefore, we ended up having 12 students for TENSS 2015:

1.
2.
3.
4.

Student name

Affiliation

E-mail

Clin,
Alexandru
Carnevale,
Federico
Eichler,
Ronny
Fabiszak,
Margaret

Department of Pharmacology,
University of Oxford, UK

alexandru.calin@sjc.ox.ac.uk

Cold Spring Harbor Laboratory, USA

fcarneva@cshl.edu

5.

Horn,
Henrique

6.

Jha, Urvashi

7.
8.
9.
10.
11.

Lee, KuoSheng
Plattner,
Viktor
Robacha,
Magdalena
SanguinettiScheck, Juan
Ignacio
iriac,

Donders Centre for Neuroscience,

Radboud University, Netherlands
Laboratory of Neural Systems,
Rockefeller University, USA
Edmond & Lily Safra Center for Brain
Sciences, The Hebrew University
Jerusalem, Israel
Neurobiology, National Centre for
Biological Sciences, India
Max Planck Florida Institute for
Neuroscience, USA
Institute of Experimental Medicine,
Budapest, Hungary
Nencki Institute of Experimental
Biology, Warsaw, Poland

ronny.eichler@gmail.com
mfabiszak@rockefeller.edu
Henrike.Horn@mail.huji.ac.il
urvashij@ncbs.res.in
Kuo-Sheng.Lee@mpfi.org
plattner.viktor@koki.mta.hu
mrobacha@nencki.gov.pl

Bernstein Center for Computational

Neuroscience, Berlin, Germany

juan.sanguinetti@bccnberlin.de

The University of Iowa, USA

alexandre-tiriac@uiowa.edu
4

Alexandre
12.

Weir, Peter

Biology and Biological Engineering,

California Institute of Technology,
USA

peter.weir@gmail.com

Table 1. Students participating in TENSS 2015 (alphabetic order)

Due to the experimental nature of TENSS where students have to get a direct hands-on
experience, only a limited number of places was made available. Students were grouped in 4
teams that worked on 4 independent setups running in parallel during lab sessions. By having
small groups at any given setup, the individual access to laboratory equipment was
maximized. The small group structure also ensured that each student had full exposure to
various techniques taught during the course and acquired the relevant experimental skills.

2.3. General structure of the program

The program of TENSS 2015 included typically 3-4 lectures per day, interleaved with several
lab sessions. Lectures were limited to 1 hour and an additional 15 minutes for questions. Lab
sessions usually took several hours and could continue until late in the night. Students directly
interacted with lecturers and had plenty of opportunities to discuss, ask questions, and get
first hand experience from them in the labs. Experienced teaching assistants provided
rigorous experimental training to students during lab sessions. TENSS covered a broad list of
topics focusing on experimental and computational methods currently employed in the field
to understand the function of neural circuits in the mammalian brain. The course lectures and
laboratory sessions included: modern optical, optogenetic and electrophysiological recording
techniques in anesthetized as well as awake (behavior) rodents, zebrafish, and flies, data
analysis, in depth lectures on specific neural circuits and direct applications, as detailed
below. The new elements introduced in 2015 compared to previous editions are marked in
red.
Optics
1. Basic Optics Diffraction, Lenses, Resolution, Illumination
2. Fluorescence & Noise Measurements
3. Skywatch (stars & beads, telescopes & microscopes)
4. In vivo Intrinsic Optical Imaging, Widefield Fluorescence
5

5. Scanning Microscopy Confocal, 2p build, code, use two lasers 2 customs & 2
commercial setups
6. Labview, Arduino, Python coding
7. Electronics & Arduinos
8. Light Activated Channels and Pumps - optogenetics
9. Adaptive optics
10. Miniaturized (headmounted) microscopes
11. Patterned Illumination techniques DLP, SLM (holography)
Electrophysiology & Behavior
12. Extracellular Recordings multiunits, single units
13. Open ephys, Tetrode & drive making & implanting
14. In vivo Patching blind & 2p guided
15. Behavior & Circuit Manipulation philosophy and tricks of flies, fish & mice
16. Virtual reality & closed loop environments
17. Machine vision
18. Bonsai
19. Sensors, actuators, head-fixed behaviors (Sanders boxes)
20. Spike sorting ephys analysis
21. Synchrony & Oscillations
22. Student projects

2.4. Lab sessions

Lab sessions are the most important component of TENSS, and take place in several dedicated
rooms at Pike Lake. Many independent rigs have been operated in parallel, including custom
optical setups for optic bench exercises, microscopes (widefield fluorescence, intrinsic
imaging, 2-photon laser scanning), surgery equipment (anesthesia, drills, surgery
microscopes), several electrophysiology setups (patch clamp, custom-built tetrode recording,
tetrode manufacturing, chronic multi-tetrode / optogenetic headstages implanted with
stereotaxic guidance for behaving mice, and so on). The students also built custom electronic
circuits to learn principles of signal acquisition and understand basic principles of
electrophysiology in a dish and design custom software to control galvanometric scanners
and video cameras using National Instruments and Digilent boards and Labview. Lab sessions
6

were organized by experienced teaching assistants (TAs) that helped & taught the students
throughout the duration of the course: Priyanka Gupta and Mehrab Modi from Cold Spring
Harbor Laboratory (CSHL), Balzs Hangya from CSHL and Institute for Experimental Medicine
in Budapest, Hungary, Petr Znamenskyi and Rob Campbell from Biozentrum Basel, Josh
Sanders from Danish Research Institute of Translational Neuroscience in Aarhus Denmark,
Dreosti Elena from University College London, Gonalo Lopes and Joana Maria Marques
Nogueira from Champalimaud in Lisbon, Adriana Dbcan and Vasile Vlad Moca from RIST
Coneural, and Ovidiu Jurju from the Centre for Integrative Neuroscience in Tbingen.
Chronologically, lab sessions covered the following:
1. A first set of labs was concerned with basic optics, lenses, custom building simple
microscopes, Koehler illumination. Students learned how to construct microscopes
themselves on the horizontal, on an optical rail by using lenses and accessories (blue
LEDs, irises, lens holders, posts, post holders, laser pointers, CCD and CMOS cameras)
bought from the course budget, or obtained on loan from companies participating in
TENSS, and used their custom setups to image samples ranging from their own hair to
Golgi stains in fixed rat brain slices. Each group had to present images that they
acquired with their custom microscope.
2. In a second set of labs, students learned to build custom fluorescence microscopes
relying on knowledge acquired in the previous lab session, transforming the bright
light microscopes into fluorescence rigs. Custom setups were used to image
fluorescent beads, pollen grain, fluorescent paper and fixed brain slices. Students
were next taught to identify and measure noise sources in their setups (dark noise,
read out noise, shot noise) and to use image analysis methods. Also, commercial
Olympus and Scientifica microscopes were used for wide-field fluorescence to
determine noise and point-spread functions by comparison.
3. The next set of lab sessions were concerned with intrinsic optical imaging, whereby
students needed to build custom rigs for intrinsic imaging and recorded in vivo data
from the olfactory system from anesthetized animals receiving olfactory stimulation
(EMX-Cre x Ai38-GCaMP3 mice). The microscopes were built from custom parts
(Nikkor SLR lenses, cameras from Allied/Vosskuhler and Photonfocus controlled by NI
boards with Labview) and were used both for fluorescence and intrinsic imaging of
brain slices and in vivo. An odor delivery machine was assembled to stimulate the
7

olfactory system of transgenic mice and results were compared across intrinsic and
fluorescence techniques (imaging GCaMP3 responses in the glomerular layer of the
bulb).
4. In parallel with the intrinsic and fluorescence microscopy, students received also
training in Labview, Matlab, C#, Python, Arduino and Bonsai with extended sessions
teaching them basic programming rules and preparing them to program the scanners
for 2-photon imaging. This years addition of a dedicated module on electronics and
micro-controllers (Arduino) was extremely popular with the participants.
5. With the next set of labs, individual groups started to work on different rigs/setups.
Two-photon (2p) microscopy was now done in parallel in two independent custom
setups and students used various parts to build their own 2p microscope from scratch.
Demos were made also with two commercial microscopes: the MOM from Sutter and
another microscope from Scientifica.
6. When the custom 2p microscopes were ready they were used to acquire 2p images
from fixed slices as well as in vivo preparations in transgenic mice, flies and zebra fish.
Images were compared to those obtained by the commercial Sutter and Scientifica 2p
setup that TENSS received on loan. Furthermore, the students were demonstrated the
usage of electrical tunable lenses (ETLs) as a promising alternative to fast z-scanning
strategies currently available.
7. Next, the electrophysiology module began, whereby students first learned how to
design custom electronic signal amplifiers by using electrical breadboards and various
electronic parts (operational amplifiers, resistors, transistors, diodes, capacitors and
so on).
8. In the next lab sessions, participants learned electrophysiology in a dish of saline and
agarose. They went through demonstration experiments relating position of electric
field dipoles and signals acquired by electrodes. Participants were also taught about
basic principles of extracellular recordings, how to make tetrodes and design and
mount microdrives and fiber optics and headstages for behavioral experiments, etc.
9. The next day was dedicated to monitoring animal behavior and in vivo patch clamping.
To monitor animal behavior they learned to write small programs in Labview and
Bonsai that used input from CMOS cameras and also concepts related to state
machines to describe the trajectory of the animal in time in open or closed loop. The
8

in vivo patch clamp demonstrations allowed students to see and also do themselves in
vivo patching. The evening was used to prepare the behavioral and acute setups for
the following days and students could witness how various parts have to be assembled
to obtain functional setups. In addition, we included also some more intensive
modules for teaching students how to program, the sessions being taught in an
interactive fashion.
10. The next lab sessions dealt with monitoring animal behavior and in vivo extracellular
recordings with optogenetics from freely moving mice. Mice were implanted with
several tetrodes 10 days before the lab sessions started by the Teaching Assistants in
the medial prefrontal cortex and striatum and were trained to follow a food pellet.
The monitoring of animal behavior was used in parallel with recording from these
behaving mice and also causally interfering with brain activity using optogenetics.
11. Recording sessions were complemented with extensive sessions for training in data
analysis. Students analyzed the very data they recorded from the behaving mice with
the analysis techniques they learned in those sessions. They wrote their own custom
software in Matlab/Python for spike sorting.
12. In addition, an important emphasis was on teaching students how to design their
custom behavioral paradigms, which they then applied directly during the recording
sessions.
13. With the help of Georg Keller, a virtual reality setup was custom built from parts
available in warehouses around Pike Lake. The VR setup using Labview and Python
scripts worked very well and students could test the head-fixed, mouse on a ball,
recording paradigm.
14. Finally, students had the chance to go more in depth with techniques that were most
interesting to them and/or that were still unclear.
15. The last three days were dedicated to student projects, whereby participants could
propose and carry out short projects of their own choice, working together in small
informal groups and applying what they have learned during the school. These last
days were the most creative ones, with projects ranging from tracking and recording
from ants, to in vivo 2p-guided patching, closed-loop experiments in behaving mice,
ants and semaquatics bugs of Pike Lake, or even wide-field fluorescence imaging of
neocortex through thinned skull in a mouse performing a complex task.
9

Lab sessions were particularly exciting and intensive, but also quite exhaustive for participants
and teaching assistants. Frequently, labs continued well beyond midnight, showing the
dedication and excitement of students. For these reasons, the program also included break
days (1 full day and 2 half days) with trips through Transylvania such that participants could
relax and recover after the exhausting activities in the labs.

2.5. Lectures
Apart from intense experimental training, participants also received theoretical lectures from
top-level scientists and had the opportunity for a close interaction with these, sprouting
interesting scientific debates. The following instructors and TAs gave presentations on various
topics covered by the summer school:
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
21.
22.
23.
24.
25.
26.
27.
28.
29.

Lecturer/TA name
Institution
Acsdy, Lszlo
Institute for Experimental Medicine, Hungary
Albeanu, Florin
Cold Spring Harbor Laboratory, USA
Battaglia, Francesco Radboud University Nijmengen, Netherlands
Bhalla, Upinder
NCBS, India
Burrone, Juan
Kings College London, UK
Campbell, Alexander Biozentrum Basel, Switzerland
Dbcan, Adriana
Romanian Institute of Science and Technology, Romania
Dreosti, Elena
University College London, UK
Engert, Florian
Harvard University, USA
Faraon, Andrei
Caltech, USA
Grdinaru, Viviana
Caltech, USA
Gupta, Priyanka
Institute for Experimental Medicine, Hungary
Hangya, Balzs
Cold Spring Harbor Laboratory, USA
Husser, Michael
University College London, UK
Hofer, Sonja
Biozentrum Basel, Switzerland
Hromdka, Tom
Cold Spring Harbor Laboratory, USA
Hbener, Mark
Max Planck Institute, Martinsried, Germany
Ji, Na
HHMI Janelia Farm, USA
Kampff, Adam
University College London, UK
Keller, Georg
Friedrich Miescher Institute, Switzerland
Lopes, Gonalo
Champalimaud Foundation, Institute for the Unknown, Portugal
Modi, Mehrab
National Centre for Biological Sciences, Bangalore, India
Monyer, Hannah
University of Heidelberg, Germany
Mrsic-Flgel, Tom
Biozentrum Basel, Switzerland
Murean, Raul
Romanian Institute of Science and Technology, Romania
Sanders, Josh
Danish Research Institute of Translational Neuroscience, Denmark
Singer, Wolf
Max Planck Institute for Brain Research, Germany
Whitlock, Jonathan
Kavli Institute for Systems Neuroscience, Norway
Znamenskyi, Petr
Biozentrum Basel, Switzerland

Table 2. Instructors at TENSS 2015

10

Lectures were held in the lecture room allocated to TENSS and had coffee breaks that
stimulated more informal questions and interaction between lecturers and students. Overall,
students were very interested and asked many excellent questions such that TENSS
organizers received very good remarks about the students from the instructors. Worth noting
is also the fun part of the lectures and interaction with lecturers. While in previous year
Florian Engert gave his lectures by the pool, this year he lectured on the hills surrounding the
pension, at a barbecue, using only a whiteboard. During past editions, the interaction
between students and professors sprouted ensuing collaborations, several of the students
going to either PhD or postdoc in labs of TENSS lecturers. Not only students and lecturers
interacted but also lecturers among themselves. Therefore, TENSS is an excellent opportunity
for networking and to discuss science.
Lectures covered a wide range of the important topics that were studied in the labs (see also
section 5). More precisely, after the opening lecture, Florin Albeanu, Priyanka Gupta, and
Adriana Dbcan started with basic principles related to optics (nature of light, lenses,
microscopes, Koehler illumination, Resolution, and so on). Next day, Priyanka Gupta made a
recap on diffraction, resolution, numerical aperture, and objectives, while Florin Albeanu
discussed wide-field fluorescence imaging, general problems related to fluorescence
(bleaching, ratiometry, and so on) and Adam Kampff lectured on detectors (cameras,
photodiodes, photomultipliers, with a brief foray into transistors MOSFET & CMOS etc) and
noise.
Then, Juan Burrone covered fluorescent probes (GFP, calcium indicators, voltage dyes,
synaptophluorins etc) and Mark Hbener discussed wide-field intrinsic optical imaging in
comparison with wide-field fluorescence imaging. Adam Kampff presented principles behind
confocal and 2-photon microscopy on day 5. On the sixth day Adriana Dbcan and Petr
Znamenskyi covered general aspects related to optogenetics and their applications and on
day seven Tom Mrsic-Flogel lectured on 2-photon microscopy (the theory behind it and its
applications). On the next day, Florin Abeanu and Michael Husser presented one photon
(intensity, DMD-based) and multi-photon patterned illumination (phase modulation),
respectively. Na Ji from HHMI Janelia Farm discussed adaptive optics, on day 9, and Sonja
Hoffer showed how to combine optical and electrophysiological methods in order to map
11

functional connectivity in brain circuits, on day 10. On the same day, Florian Engert gave a talk
on LASERS on a hilltop close to Pike Lake, during an informal barbecue. The behavior module
has been started on day 11 with a lecture by Adam Kampff and electrophysiology began with
a talk by Upi Bhalla on biophysics of neurons and a lecture by Tomas Hromadka in the evening
on in vivo patch clamp. On day 12, Wolf Singer gave a more theoretical lecture on synchrony,
oscillations and neural ensembles and Petr Znamenskyi discussed chronic multi-electrode
extracellular recordings. Balzs Hangya started the data analysis module on day 13 and this
was followed by a lecture of Hannah Monyer on inhibition and a crash-course on neural
oscillations by Raul Murean. Day 14 was dedicated to virtual reality by Georg Keller and
Gonalo Lopes. In the evening, Jonathan Whitlock discussed approaches to understand
systems exemplified by navigation, including grid cells, place cells, and head direction cells.
Lszl Acsdy then covered acute recording with intra- and juxtacellular techniques in the
thalamus and Viviana Grdinaru talked about optogenetics and CLARITY, on day 17. In the
evening, Francesco Battaglia discussed large-scale array recordings. The last day that had
theoretical lectures was on June 18, when Gonalo Lopes discussed in depth concepts related
to behavioral design balancing the advantages and disadvantages of controlled behaviors in
the lab vs. open arenas/open field monitoring. Danko Nikoli, Venki Murthy and Simon
Rumpel could not make it to the school and cancelled in the last moment, due to unforseen
problems.

12

2.6. Some pictures from TENSS 2015

Intense preparations for TENSS, before the school starts. Top: preparing the tables for the
2-photon rigs. Bottom: surgery room, implanting mice with chronic drives.
13

Rob Campbell, assembling his telescope

Priyanka Gupta and Toma Hromadka preparing all that is necessary for the optical tabletop exercises that will happen in the first days of the course
14

Opening day of TENSS 2015 Adam Kampff, Florin Albeanu & Raul Murean

Florin and Adam explaining the TENSS spirit

15

The students...

What is light? Florin delivering the first lecture of the course.

16

Fiddling with lenses Petr Znamenskiy

Rob, Toma and Iuliu Vasilescu showing students how light is focused through lenses
17

Ghosts?

Many lenses with Priyanka

18

A working horizontal microscope

Focusing with Petr

19

First student presentations

Juan Burrone, lecturing on fluorescent voltage probes

20

Fluorescence images recorded with microscopes built by students

And Arduinos..
21

Rob discussing astronomy a good primer for optics principles

Flowers around Pike Lake

22

Playing volley with an improvised net

And dancing...
23

Playing football...

Local children learning how to harvest the hay

24

Adam lecturing...

Several custom built intrinsic imaging rigs, with guidance from Mark Hbener
25

Watching the UEFA Champions League final (Barcelona Juventus). From left to right:
Michael Husser, Tom Mrsi-Flogel, Josh Sanders and Mark Hbener

Students just about to start building custom 2-photon microscopes. In the foreground:
already working commercial microscopes from Sutter and Scientifica
26

Making progress with the custom 2-photon setup

27

Aligning the microscopes

With help from friends... From left to right: Georg Keller, Mark Hbener, Tom Mrsi-Flogel,
and Florian Engert.
28

Na Ji lecturing about adaptive optics

First images from a custom 2-photon microscope

29

And Toma preparing the in vivo patching using the Axopatch and the Scientifica Slicescope

A science chat with Florin, Tom, Adam and Sonja Hofer

30

Tuning the custom 2-photon microscope

Na Ji again lecturing on adaptive optics

31

Josh preparing a multisensory head-fixed 3D printed behavioral setup

The LASER room with two functional custom-built 2-photon microscopes (left) and two
commercial ones (right)

32

Lunch time

An the beautiful barbecue, with students preparing the lamb (South American style)
33

Florian Engert teaching lasers on the hills around Pike Lake

34

Transylvanian countryside with sheep contemplating the TENSS barbecue...

Florian and Georg relaxing after the barbecue and lecture on the hills
35

And the ephys and behavior module begins...demo-ing a miniaturized head-fixed

microscope from Doric Lenses

Cell in dish learning basics of electrophysiology

36

Upi Bhalla teaching biophysics of neurons

Adam again...
37

Wolf Singer and Florian Engert

Gary Wilson from Gatsby Foundation discussing with Georg Keller

38

Toma on the patch clamp techniques

Florian again lecturing on Electron Microscopy

39

And Adam...

Tom testing artificial stereovision with a VR headset (OCULUS)

40

Learning how to build headstages for electrophysiology

And patch clamp with Toma

41

Wolf Singer on hard problems in neuroscience

And Petr, explaining electrophysiology recording with tetrodes

42

Gone swimming in the lake

43

A custom-built VR setup for mice that just works

Toma again, about programming rules

44

Raul, working on this report...

Florin preparing tools for a surgery

45

Wolf Singer in action

Teaching the bubble sort algorithm by ordering students according to their height...

46

Toma preparing the 2-photon targeted in vivo patching with Mehrab Modi

47

Francesco Battaglia and Raul in an ad hoc debate about oscillations in the brain

48

Lszl Acsdy lecturing about the thalamus

Viviana Grdinaru on CLARITY

49

Two-photon targeted patching in vivo

Mehrab and Priyanka finetuning

50

Students recording their behavioral data for their final group project

51

Final student presentations

52

Group pictures TENSS 2015

53

2.7. Contribution of TENSS to Neuroscience and future directions

The main objective of TENSS is to provide top experimental training to graduate students
from all over the world. Although the organizational effort for this type of summer school is
tremendous, it pays off because in about three weeks the school exposes students to such a
wide range of experimental techniques that sometimes require months or years and
switching between several labs to learn. Therefore, we believe that TENSS brings an
important contribution to the field worldwide and mostly to Neuroscience in Europe and
Romania where such experimental intensive schools were until recently missing. In addition, a
top-level experimental school helps putting Romania on the world map of excellent training
centers in neurosciences.
We intend to make TENSS a tradition and have it organized on an annual basis. Already at its
fourth edition, TENSS has a promising outlook and we are already taking steps to secure
funding for 2016. The school had important echoes in the international and national press
and we present two examples:
In Nature:
http://www.nature.com/news/summer-skills-1.15416
On Digi24 news TV channel:
https://www.youtube.com/watch?v=wCPAl_vlY0g

54

3. Information regarding the organizing of the event

Organizing TENSS requires months of intensive preparations. The organizing team for the
2015 edition consisted of 15 people that worked around the clock, pro bono, to make TENSS
happen. In addition, several external factors also contribute to the success of TENSS. For
example, several large companies that produce scientific equipment are involved by providing
equipment for free, on loan, to be used during the course. TENSS 2015 benefited from the
following equipment:
Optical bench equipment (lenses, irises, lens tubes, posts, post holders, mirrors,
mirror holders, optical cage cubes, objectives, SLR lenses, galvanometric-scanners,
clamps, analog input/output and digital input/output acquisition boards, excitation
filters, emission filters, dichroic mirrors, photomultiplier tubes) mostly bought from
the courses budget.
Femto-second pulsed tunable laser infrared Chameleon XR90 MHz from Coherent,
received as a donation for the course and for the host institute.
A second femto-second pulsed laser (not tunable) HiQ2 laser on loan from Spectra
Physics.
A SliceScope and a 2-photon microscope manufactured by Scientifica Ltd, UK.
Intrinsic Optical Imaging and wide-field epifluorescence LED based rig (custom built).
The setup was built from SLR lenses, tubes, lens holders, LEDs, and CCD cameras
(Allied Vosskuhler) bought from the course budget.
One epifluorescence microscope based on Olympus design BX51 donated as
permanent equipment for the course by Olympus Europe.
Extracellular Recording (Tetrode Arrays) headstages and recording setup from Open
Ephys. Tetrodes were manufactures using equipment from Neuralynx received on loan
for the course.
Axopatch 200B amplifiers for the clamp setups manufactured by Molecular Devices,
USA able to record/induce action potentials and sub-threshold (synaptic) activity in
individual neurons in acute slice and/or slice culture preparations.
A pipette puller received on loan, manufactured by Narishige.
Micro-manipulators for maneuvering recording electrodes manufactured by Sutter
Instruments (MP-285 and MP-225).
55

 A MOM 2-photon microscope received on loan from Sutter Instruments and

assembled by Rick Ayer on spot.
Data acquisition and hardware control electronic boards by National Instruments.
Optical and optomechanical components and breadboards for assembling the optical
imaging setups, manufactured by Thorlabs.
Many parts were bought from the budget of the course and will be used also for future
editions of the course. Since the course started more than 500 individual parts had to be
purchased for the course.
The budget of TENSS 2015 totaled at almost 80,000 EUR, that were covered by the Gatsby
Foundation, Wellcome Trust, FENS, IBRO, The Company of Biologists, Hertie Foundation,
EBBS, and student participation fees. In addition, the course received support from labs in
Europe and USA (Florin Albeanu, Lszl Acsdi, Tom Mrsic-Flogel, Georg Keller) which
provided transgenic mice and various equipment for free. The University of Medicine and
Pharmacy in Cluj-Napoca also helped the course by loaning two surgery microscopes and
donating wild-type mice.
The notification & selection of students took several months to complete. A call for
applications was issued in January 2015 and the deadline was on March 8th. Applications
consisted of one letter of intent, one CV and two independent recommendation letters. A
board of 15 scientists rated the applications as objectively as possible in a 2-phase selection
process and notification of acceptance was sent out on April 7th. Immediately before the
beginning of the summer school, the organizing team together with company representatives
went to Pike Lake to set up the labs and prepare everything for the arrival of students. After
the conclusion of TENSS 2015, the dismantling of lab equipment and return of loan
equipment takes several additional days. Therefore, the total logistics effort for this type of
experimental summer school lasts for about 1.5 months (2 weeks before, 3 weeks the
summer school, 1 week after). This adds up to the several months necessary to raise funding
and organize the logistics of the course, including equipment delivery.
Presently, the TENSS team is finishing all activities related to TENSS 2015 and already started
to raise funds for the next edition, TENSS 2016.
56

4. Financial report
The total budget of TENSS 2015 was a very large one because of the nature of the summer
school. Experimental schools require a tremendous logistic effort and lots of equipment and
therefore costs are much higher than in schools which are limited to theoretical lectures only.
The total spending of TENSS 2015 was around 78.120 EUR as of June 24th, 2015. The budget
structure of TENSS 2015 is outlined in the following table.
Nr.
crt.
1.
2.
3.
4.

Budget category
Accommodation & meals
Travel costs
Logistics***
Equipment
Total

Cost (RON)

Cost (EUR*)

Cost (GBP**)

115542
66371
66459
96138
344510

26200
15050
15070
21800
78120

18602
10686
10700
15478
55466

Table 3. Spending per budget categories (rounded figures). *The costs in EUR were
estimated using the average exchange rate of ING Bank Romania for the corresponding
expenditure period (4.41 EUR/RON). **The costs in GBP were computed by taking an average
EUR / GBP exchange rate of 0.71. ***Logistics costs are estimates as of 24th June 2015,
because there are additional costs for shipping loaned equipment back which, on this date,
could only be estimated.
The sources of funding are described as follows.
Funding source

Amount
(EUR)
25000
25000
5000
5000

1.
1.
2.
3.

Gatsby
Wellcome Trust
FENS
IBRO

4.

The Company of Biologists

3900*

5.
6.
7.

Hertie Foundation
EBBS
Registration fees
Total

3000
700
10500
78100

Amount
(GBP)
17750
17750
3550
3550
2769
(3000)
2130
497
7455
55451

% Budget
32.01%
32.01%
6.40%
6.40%
5.00%
3.84%
0.90%
13.44%
100%

Table 4. Funding sources that covered the budget for TENSS 2015. * - the actual amount
received in the institutes account.
57

5. Final program of the event

The schedule of TENSS 2015 included each day several lectures and lab sessions, with breaks
in between.

58

TENSS 2015 Schedule

Color code:
DAY

TIME SLOT

DAY 1

08:00 09:00

General

Lectures

Labs

Student presentations

MAIN RESPONSIBLE

GROUPS

Breakfast

09:00 10:00

Florin, Raul and Adam

10:00 11:15

Florin Albeanu

Introduction to the course

Intro to Optics

11:15 11:30
11:30 13:00

Coffee break
Adriana Dabacan

Image formation with Lenses

13:00 14:00
14:00 17:00

Lunch
Adriana, Petr, Rob and Priyanka

ABCD

17:00 17:30

Simple microscopes : Lenses and image formation properties

Coffee

17:30 18:30

Priyanka Gupta

18:30 20:30

Adriana, Petr, Rob and Priyanka

20:30 21:30

CONTENT

Koehler Illumination and Numerical aperture

ABCD

Bench-top koehler microscopes depth of field and aperture

Dinner

21:30 23:00
DAY 2

08:00 09:00

Morning run/swim

09:00 10:00

Breakfast

59

10:00 11:15

Priyanka Gupta

Recap diffraction, resolution, numerical aperture, objectives

11:15 11:30
11:30 13:00

Coffee break
Fluorescence: Wide-field epi-fluorescence, PSFs and resolution, dF/F,
bleaching, ratiometry (dF/dR)

Florin Albeanu

13:00 14:00
14:00 16:00

Lunch
Detecting signals: Noise, Cameras, PMTs and diodes, Introducing lab
session on noise measurements

Adam Kampff

16:00 16:15
16:15 18:00

Coffee break
Adriana, Petr, Rob and Priyanka

ABCD

18:00 18:15
18:15 20:30

Coffee break
Adriana, Petr, Rob and Priyanka

ABCD

20:30 21:30
21:30 23:00
DAY 3

Convert bench-top koehler microscopes to epi-fluorescence

Noise measurements, measure PSFs using fluorescent beads

Dinner

Adriana, Petr, Rob and Priyanka

ABCD

Discussion, analysis, continue Labs

08:00 09:00

Morning run/swim

09:00 10:00

Breakfast

10:00 11:30

Fluorescent probes: GFP, calcium indicators vs. voltage dyes,

synaptophluorins

Juan Burrone

11:30 11:45
11:45 13:00

Coffee break
Josh & Rob

Arduinos & Electronics Part I

13:00 14:00
14:00 17:00

Lunch
Adriana, Petr, Rob and Priyanka

ABCD

Image fixed brain slices on the bench-top fluorescence microscopes,

60

Compare noise and PSFs on commercial Scientifica and Olympus

microscopes (1-2 students per group can combine and do this together),
In parallel, all groups analyze noise and PSF measurements, compile
results and make presentations.
17:00 17:30
17:30 20:30

ABCD

Acquire images of fluorescent samples - fish, slices etc. Continuation of

analysis and making presentations.
Dinner

21:30 23:00

Student presentations (15 + 5 minutes): PSFs, noise characterization of

wide-field microscopes
Rob and Adam

Skywatch

08:00 09:00

Morning run/swim

09:00 10:00

Breakfast

10:00 12:00

DAY 5

Adriana, Petr, Rob and Priyanka

20:30 21:30

23:00 onwards
DAY 4

Coffee break

Josh & Rob

Arduinos & Electronics Part II

12:00 19:30

Free time to relax

19:30 onwards

Dinner and party

08:00 09:00

Morning run/swim

09:00 10:00

Breakfast

10:00 11:30

Mark Hubener

11:30 11:45
11:45 13:00

Intrinsic Imaging - Principles and Intro to Lab session - practical aspects

and comparison with wide field fluorescence imaging
Coffee break

Adam Kampff

Scanning and Confocal Microscopy; Intro to Lab session on scanning

software

61

13:00 14:00

14:00 16:00

Lunch

Adriana, Petr, Rob and Priyanka

AB

Set up microscopes for intrinsic and widefield fluorescence imaging and

determine optimal imaging parameters

CD

Set up bench top scanners and simple beam alignment

16:00 16:30
16:00 20:30

Coffee break
Adriana, Petr, Rob and Priyanka

AB

Image intrinsic optical and fluorecence signals, analyze acquired signals

CD

Write scanning software

20:30 21:30
21:30 23:00
DAY 6

Dinner
Adriana, Petr, Rob and Priyanka

ABCD

Continue respective lab sessions

08:00 09:00

Morning run/swim

09:00 10:00

Breakfast

10:00 12:00

Adriana, Petr, Rob, Mehrab and Priyanka

AB

Set up bench top scanners and simple beam alignment

CD

Set up microscopes for intrinsic and widefield fluorescence imaging and

determine optimal imaging parameters

12:00 13:00
13:00 17:30

Early Lunch
Adriana, Petr, Rob, Mehrab and Priyanka

AB

Write scanning software

CD

Image intrinsic optical and fluorecence signals, analyze acquired signals

17:30 18:00
18:00 19:30
19:30 20:30

Coffee break
Adriana, Petr, Rob, Mehrab and Priyanka

ABCD

Continue respective lab sessions

Early Dinner

62

20:30 22:00
22:00 - 23:30
DAY 7

ABCD
Adriana, Petr

Introduction to optogenetics and applications

08:00 09:00

Morning run/swim

09:00 10:00

Breakfast

10:00 11:00

Tom Flogel

Two-photon microscopes the THEORY and applications in neuroscience

11:00 11:15

11:15 13:00

Coffee break
Adam Kampff

AB

Write X-Y- Z scanning and image acquisition software

Florin Albeanu

CD

Building a two-photon microscope general discussion on practical

aspects and brief overview of commercially available setups

13:00 14:00
14:00 17:00

Lunch
Petr, Rob, Adam, Goncalo and Elena

AB

Write X-Y- Z scanning and image acquisition software

CD

Build your own two-photon microscope

17:00 17:30
17:30 20:30

Coffee
Petr, Rob, Adam, Goncalo and Elena

ABCD

20:30 21:30
21:30 23:00
DAY 8

Student presentations (15 + 5 minutes): Widefield imaging

Continuation of lab sessions

Dinner

Petr, Rob, Adam, Goncalo and Elena

ABCD

Continuation of lab sessions

08:00 09:00

Morning run/swim

09:00 10:00

Breakfast

10:00 10:30

Florin Albeanu

One Photon patterned illumination - Intensity modulation

10:30 11:30

Mike Hausser

Multiphoton patterned illumination - Phase modulation

63

11:30 11:45

11:45 13:00

Coffee break
Florin Albeanu

AB

Building a two-photon microscope general discussion on practical

aspects and brief overview of commercially available setups

Adam Kampff

CD

Write X-Y- Z scanning and image acquisition software

13:00 14:00
14:00 17:00

Lunch
Petr, Rob, Adam, Goncalo and Elena

AB

Build your own two-photon microscope

CD

Write X-Y- Z scanning and image acquisition software

17:00 17:15
17:15 20:30

Coffee
Petr, Rob, Adam, Goncalo and Elena

ABCD

20:30 21:30
21:30 23:00
DAY 9

Continuation of lab sessions

Dinner

Petr, Rob, Adam, Goncalo and Elena

ABCD

Continuation of lab sessions

08:00 09:00

Morning run/swim

09:00 10:00

Breakfast

10:00 - 11:00

Na Ji

11:00 13:00

Mehrab N. Modi

13:00 14:00
14:00 17:00

20:30 21:30

in vivo 2p Lab imaging session

Lunch

Mehrab N. Modi

17:00 17:30
17:30 20:30

Adaptive Optics

in vivo 2p Lab imaging session

Coffee

Mehrab N. Modi

in vivo 2p Lab imaging session

Dinner

64

21:30 23:30
DAY 10

Demo of miniature head-mounted microscopes / make presentations

08:00 09:00

Morning run/swim

09:00 10:00

Breakfast

10:00 11:00

DAY 11

Georg Keller

Combining optical and ephys approaches to study functional connectivity

and tuning of neuronal circuits

Sonia Hofer

11:00 13:00

Student presentations - multiphoton microscopy

13:00 20:30

Picnic/Barbecue on the hills featuring Florian and LASERS

20:30 onwards

Dinner and Party

08:00 09:00

Morning run/swim

09:00 10:00

Breakfast

10:00 11:30

Adam Kampff

Behavior module, clicker training and machine vision

11:30-11:45
11:45 13:15

Coffee break
Biophysics of neurons - RC circuits, dipoles and impedance (Introduce
'cell in a dish' lab demo)

Upi Bhalla

13:15 14:15

14:00 17:00

Lunch
Upi and Mehrab

AB

Cell in a dish: Bench top electronics and basics of electrophysiology

Adam and Elena

CD

Animal behavior I - basic principles, clicker training, introduction to

machine vision

17:00 17:30
17:30 20:30

Coffee break
Adam and Elena

AB

Animal behavior I - basic principles, clicker training, introduction to

machine vision

65

Upi and Mehrab

CD

20:30 21:30
21:30 23:00
DAY 12

Cell in a dish: Bench top electronics and basics of electrophysiology

Dinner

Tomas Hromadka

Introduction to patch clamp

08:00 09:00

Morning run/swim

09:00 10:00

Breakfast

10:00 11:00

Wolf Singer

11:00 12:00

Petr

Time as coding space in cortical processing

Introduction to chronic extracellular recordings

12:00 13:00
13:00 14:00

14:00 16:00

Early Lunch
AB

Tetrode making

CD

Building Tetrode drives

Tomas Hromadka

AB

In vivo patch clamp

Balazs, Josh and Petr

CD

Building Tetrode drives

Balazs, Josh and Petr

16:00 16:15
16:15 17:15

17:15 19:15
19:15 - 20:30
20:30 21:30

Coffee break
AB

Building Tetrode drives

CD

Tetrode making

Balazs, Josh and Petr

AB

Building Tetrode drives

Tomas Hromadka

CD

In vivo patch clamp

Balazs, Josh and Petr

Tomas, Balazs, Josh, Petr

Tetrode making, drive building, patching contd..

Dinner

66

21:30 23:00
DAY 13

Tomas, Balazs, Josh, Petr

ABCD

Tetrode making, drive building, patching contd..

08:00 09:00

Morning run/swim

09:00 10:00

Breakfast

10:00 13:00

Balazs, Josh and Petr

AB

Physiology I - familiarize with set-up and implanted animals, record

single cell and local field data

Goncalo

CD

Animal behavior II - machine vision

13:00 14:00

14:00 17:00

Lunch
Goncalo

AB

Animal behavior II - machine vision

Balazs, Josh and Petr

CD

Physiology I - familiarize with set-up and implanted animals, record

single cell and local field data

17:00 17:30

Coffee break

17:30 18:30

Balazs Hangya

18:30 20:30

Balazs, Josh and Petr

20:30 21:30

DAY 14

Introduction to data analysis (filtering, spike sorting, PSTH)

ABCD

Analysis of acquired data (filtering, spike sorting, PSTH)

Dinner

21:30 22:30

Hannah Monyer

22:30 23:30

Raul Muresan

Inhibition in the brain: from individual neurons to complex systems

effects
Correlations and local field potentials

08:00 09:00

Morning run/swim

09:00 10:00

Breakfast

10:00 11:00

Georg Keller

11:00 12:00

Georg, Goncalo

Virtual reality and closed loop behaviors

Virtual reality demo

67

12:00 13:00

13:00 16:00

Early lunch
Balazs, Josh and Petr

AB

Physiology II - recording from freely moving mice, optogenetic

stimulation, spike sorting

Goncalo, Josh

CD

Animal behavior III - Introduction to sensor/actuator modules

16:00 16:30
16:30 19:30

Coffee break
Goncalo, Josh

AB

Animal behavior III - Introduction to sensor/actuator modules

Balazs, Josh and Petr

CD

Physiology II - recording from freely moving mice, optogenetic

stimulation, spike sorting

19:30 20:30

ABCD

20:30 21:30
21:30 22:30

Dinner

ABCD

Student Talks

08:00 09:00

Morning run/swim

09:00 10:00

Breakfast

10:00 20:30

Trip to Cluj and Turda

20:30 onwards
DAY 16

Place cells, grid cells, head direction cells. Approaches to understanding

systems exemplified by navigation

Jonathan Whitlock

22:30 24:00
DAY 15

Continue lab/analysis/presentation making

Dinner and Party

08:00 09:00

Morning run/swim

09:00 10:00

Breakfast

10:00 12:00

Tomas

Programming principles - interactive session

68

12:00 13:00
13:00 13:45

Early lunch
Danko Nikoli

Closed-loop experimentation and adaptability of the nervous system

13:45 14:00
14:00 16:15

16:15 20:30

Coffee break
Florin and the TAs

AB

Student Project design and setup

Tomas

CD

In vivo patching

Florin and the TAs

CD

Student Project design and setup

Tomas

AB

In vivo patching

20:30 21:30
21:00 23:00
DAY 17

Dinner
Florin and the TAs

Student Project design and setup

08:00 09:00

Morning run/swim

09:00 10:00

Breakfast

10:00 11:00

Laszlo Acsady

11:00 12:00

Francesco Battaglia

Acute recording, cell type identification (juxta/tagging), anatomy.

Approaches to understanding circuits examplified by thalamic networks.
Large-scale array recordings, and using them to understand the
hippocampus-prefrontal pathway

12:00 13:00

Lunch

13:00 18:00

Group Experiments

18:00 20:30

Group Experiments

20:30 21:30

Dinner

21:00 23:00

Group Experiments

69

DAY 18

DAY 19

08:00 09:00

Running around

09:00 10:00

Breakfast

10:00 11:00

Goncalo

11:00 12:00

Viviana Gradinaru

Behavior: Experimental control and ethological relevance

Clarity talk with demo samples

12:00 13:00

Lunch

13:00 18:00

Group Experiments

18:00 20:30

Group Experiments

20:30 21:30

Dinner

21:00 23:00

Student presentations - group projects

08:00 09:00

Morning run/swim

09:00 10:00

Breakfast

10:00 11:45

Informal chalk board talks by students (10 + 5 minutes): Very brief intro
to current research work and defend future proposals to use knowledge
acquired at the course

11:45 12:00

Coffee

12:00 13:30

Informal chalk board talks by students continue

13:30 14:30

Lunch

14:30 16:00

Round up and feedback

Evening onwards

Music, movies, swim and party

70