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fluorescent
confocal
microscopy
system



1 2,3 2,3 1 1 Bertrand
Viellerobe
 ,
Isabelle
Janssens
 ,
Karine
Gombert
 ,
Hedi
Gharbi
 ,
François
Lacombe
 
and
Frédéric
D 1)
Mauna
Kea
Technologies,
9,
rue
d’Enghien,
75010
Paris,
France
 2)
CEA,
I²BM,
Service
Hospitalier
Frédéric
Joliot,
4
place
du
général
Leclerc,
91401
Orsay
(France)
 3)
INSERM
U1023,
Université
Paris
Sud,
Laboratoire
d’Imagerie
Moléculaire
Expérimentale,
4
place
du
général
Leclerc,
91401
Orsay
(France)


Multicolor Probe-based Confocal Laser Endomicroscopy Introduction

Results

3h post-injection

7

In Vivo Molecular Imaging with Cellular Resolution

Today,
confocal
fluorescence
microscopy
and
mul^photon
 Macroscopic
imaging
of
Angiostamp®
using
fDOT/CT

 microscopy
 are
 increasingly
 used
 for
 in
 vivo
 studies
 in
 The
 biodistribu^on
 of
 Angiostamp
 was
 first
 evaluated
 small
 animals.
 Such
 techniques
 allow
 studying
 the
 using
 fluorescence
 Diffuse
 Op^cal
 Tomography
 (fDOT)
 in
 structure
and
the
physiology
of
living
organism
at
cellular
 nude
 mouse
 bearing
 a
 subcutaneous
 xenogran
 tumor
 scale.
 The
 major
 limita^ons
 of
 such
 imaging
 is
 that
 1‐
 from
 NIH/MEN2A
 cells.
 This
 imaging
 technique
 has
 been
 samples
need
to
be
placed
conveniently
on
a
conven^onal
 considerably
improved
since
past
decade
and
allows
now
 microscope
 stage
 which
 require
 extensive
 surgical
 reconstruc^ng
and
quan^fying
fluorescence
signal
in
three
 prepara^on,
 and
 2‐
 rapid
 image
 collec^on
 is
 required
 to
 dimensions
 inside
 small
 animal.
 fDOT
 imaging
 fused
 with
 minimize
 the
 effects
 of
 movement
 (such
 as
 animal
 X‐Ray
 Computed
 Tomography
 (CT)
 demonstrated
 a
 high
 breathing).
 To
 solve
 this
 problem,
 novel
 confocal
 uptake
 of
 the
 tracer
 in
 the
 tumor
 area.
 Interes^ngly,
 the
 approaches
 using
 fiber
 bundle‐based
 systems
 have
 been
 Mauna Kea Technologies, Paris, France uptake
 seems
 heterogeneous
 in
 the
 tumor
 and
 seems
 developed
 by
 Mauna
 Kea
 Technologies
 (Paris,
 France).
 higher
 in
 the
 booom
 of
 the
 tumor.
 In
 subcutaneous
 Such
 systems,
 named
 Cellvizio®,
 use
 extremely
 small
 xenogran
 models,
 the
 tumour
 cannot
 easily
 grow
 to
 the
 bundles
of
fibers,
0.3–2.6
mm
in
diameter
that
can
contain
 skin
 where
 it
 cannot
 find
 a
 lot
 of
 nutrients,
 but
 it
 upwards
of
30,000
fibers.
Each
fiber
is
used
for
excita^on
 preferen^ally
 invades
 the
 ^ssue
 below.
 The
 tracer
 seems
 delivery
 and
 recovery
 of
 the
 emission
 back
 through
 the
 to
have
a
higher
uptake
in
that
zone
that
should
be
rich
in
 fiber
to
a
detector.
Hence,
each
fiber
can
be
compared
as
 new
blood
vessels.
 Perfect imaging modality doesn’t exist to date if we consider spatial resolution, sensitivity, ease of use or penetration an
 independent
 insect
 eye.
 The
 absolute
 advantages
 of
 capabilities. Conventional microscopy is widely used for in vitro and invasive studies whereas whole body imagers can’t However,
although
fDOT
can
now
detect
fluorescence
in
 this
 apparatus
 are
 size,
 flexibility,
 and
 image
 collec^on
 reach cellular details in vivo. the
nanomolar
range,
it
has
sEll
a
low
(a
few
mm)
spaEal
 speed
 (up
 to
 of
 12
 frames/s).
 Up
 to
 now,
 two
 Cellvizio®
 resoluEon
 that
 can
 not
 permit
 to
 have
 a
 precise
 idea
 of
 • Conventional microscopy systems
were
available
either
with
a
488
nm
or
a
660
nm
 the
biodistribuEon
of
the
probe
at
the
cellular
scale.
 • Requires numbers of animals: andnew
 ethicsfiber
 issues laser
 beam.
substantial Here,
 we
 describe
 the
high use
costs of
 a
 • Allows for In vitro imaging of phenomena at a given time: no dynamic imaging bundle‐based
fluorescence
imaging
prototype
(Cellvizio®
 • Provides high resolution Dual
 Band)
 that
 can
 perform
 simultaneous
 excitaEon
 • Whole body imagers MRI,nm
 CT, Optical fluorescence with
 both
 lasers
 (488
 and
 660
 nm)
 and
 recovery
 of
 • Well suited for in vivo imaging of biodistribution of molecular biomarkers emission
 signal
 with
 
 two
 detectors.
 We
 validate
 the
 • Non invasive: compatible with longitudinal studies system
 comparing
 biodistribu^on
 of
 a
resolution: fluorescent
 • Cellular information the
 unreachable due to low spatial RGD‐based
 probe
 (Angiostamp®)
 in
 different
 region
 of
 a
 CELLVIZIO® DUAL BAND is a multicolor probe-based Confocal Laser Endomicroscope tumor
xenogran
as
well
as
in
different
organs
of
a
mouse.
 • Fills the gap between conventional microscopy and whole body imagers (Fig 1 on the right) This
fluorescent
probe
is
known
to
bind
the
αvβ3
Integrin,
 • From bench to bedside: Cellvizio® is FDA approved and CE cleared for clinical indications a
protein
overexpressed
at
the
surface
of
endothelial
cells
 • Delivers dynamic in vivo fluorescence imaging of molecular events with cellular resolution (1,4 µm) during
angiogenesis
[1].
 • Can access any tissue with minimal invasiveness including deep brain, abdominal cavity, GI tract, etc...

Angiostamp®
is
surrounding
the
tumor
blood
vessels


FITC-dextran

AngioStamp ®

Angiostam

Merge

FITC-dextran

Merge

FITC-dextran

H. Gharbi*, F. Lacombe

FITC-dextran

AngioStamp ®

EMIM 2013 | TORINO | ITALY

INTRODUCTION

• Longitudinal studies made possible: evaluation of drug candidates actions on the same animal over time • Turn to Optical Biopsy and real-time diagnostics (Fig 2 below)

Materials and methods • Point Of Care molecular imaging station

Angiostamp®
is
no

Angiostamp®
is
not
surrounding
the
blood
vessels
of
muscle
 FITC-dextran

AngioStamp ®

FITC-dextran

Merge

Angiostamp®
is
not
accumulated
in
liver
 FITC-dextran

AngioStamp ®

Angiosta Merge

FITC-dextran

Angiostamp®is
not
accumulated
in
spleen
 FITC-dextran

AngioStamp ®

Angiostam Merge

Fig 1 Cellvizio bridges the gap between conventional miicroscopy and whole body imagers

FITC-dextran

Fig.
1:
BiodistribuEon
of
Angiostamp®
analyzed
by
fDOT/CT
imaging
 ●
Ethics
Statement
 Fluorescence
 signal
 reconstructed
 in
 3D
 (colored)
 was
 fused
 to
 CT All
 animal
 use
 procedures
 were
 in
 strict
 accordance
 with
 the
 
imaging
of
the
mouse
(gray).

 recommenda^ons
 of
 the
 European
 Community
 (86/609/CEE)
 and
 the
French
Na^onal
Commioee
(décret
87/848)
for
the
care
and
use
 Microscopic
 imaging
 of
 Angiostamp®
 using
 Cellvizio®
 of
laboratory
animals.
 ●
Animal
model
 Dual
Band

 Female
 nude
 mice
 (~23
 g)
 were
 subcutaneously
 injected
 with
 106
 Following
fDOT
imaging,
the
mice
were
injected
with
FITC‐ Angiostamp®
is
eliminated
by
glomerulus
of
kidney
 Angiostamp® Fig 2 Optical biopsy avoids C634Y tumor
 cells
 NIH‐MEN2A
 expressing
 the
 oncogen
 RET .
 Aner
 15
 tissue samples with
 by providing works with a large variety of fiber-optic probes that have been designed to fit with Dextran
 before
 imaging
 the
 fiber
 bundle‐based
Cellvizio® Dual Band FITC-dextran AngioStamp ® Merge FITC-dextran days,
mice
have
a
tumor
(~30‐50
mm3).

 dynamic in vivo microscopic various applications constraints, with diameters as small as a needle tip (300 µm) or with resolution that fluorescence
 imaging
 prototype
 (Cellvizio®
 Dual
 Band).
 
●
In
vivo
fluorescence
imaging
using
fDOT/CT

 images in a non orMicroscopy minimally Improvement of Fibered Fluorescence images of can reach 1,4 µm. The system is able to simultaneously track two different molecular signatures in vivo and The
instrument
allowed
to
acquired
in
real‐^me
image
of
 invasive manner. Angiostamp
 (10
 nmol)
 was
 intravenously
 injected
 into
 the
 tail
 of
 in situ, allowing therefore colocalization studies to be conducted on the go in the living animal. The individual cells in the brain of live mice Improvement of Fibered Fluorescence Microscopy images of e a l t i m ewith
 s t r u FITC‐Dextran
 cture and anesthe^zed
animals.
3D
fluorescence
images
were
acquired
3h
or
 blood
 vessels
Rlabeled
 and
 the
 signal
system’s wavelengths (488 and 660 nm) cover a large spectrum of in vivo compatible fluorescent dyes, function characterization and individual in the brain antibodies of live mice Jesus Pascual-Brazo, Veerle Reumers, Sarah-Ann Aelvoet, Zeger Debyser, Veerle Baekelandt cells proteins, biosensors, or genetically engineered animal models used routinely in translational 7h
post‐injec^on
using
a
prototype
op^cal
imager
(TomoFluo3D).
CT
 from
 Angiostamp®.
 Thanks
 to
 the
 high
 flexibility
 of
 the
 physiopathology diagnosticsFaculty is Pascual-Brazo, for Neurobiology and Gene Therapy. Department of Neurosciences. of Medicine,Veerle K.U. Reumers, Leuven Jesus Sarah-Ann Zeger Debyser, Veerle research. The Aelvoet, tremendous advances in Baekelandt biomarker discovery is putting in vivo diagnostics to a whole new imaging
 was
 perform
 using
 the
 SkyScan
 1178
Laboratory high‐throughput
 system
different
organs
can
easily
been
analyzed
as
well
as
 Laboratory for Neurobiology and Gene Therapy. Department of Neurosciences. Faculty of Medicine, K.U. Leuven therefore possible. precision level. Cellvizio® Dual Band sets the stage to a better understanding of molecular pathways that micro‐CT
 (Skyscan,
 Kon^ch,
 Belgium).
 Fusion
 of
 fDOT
 with
 CT
 was
 different
part
of
the
tumor
xenogran
(scheme
2).

 are leading to cancer, inflammation, infection or neurodegenerative diseases. performed
using
the
Brainvisa
medical
imaging
processing
sonware
 INTRODUCTION MITOCHONDRIAL REDOX STATE IN LIVE MICE INTRODUCTION MITOCHONDRIAL REDOX STATE IN LIVE MICE (hop://brainvisa.info/index_f.html)
[2].

 Imaging techniques, such as magnetic resonance imaging and Imaging techniques, such as magnetic resonance imaging and 
●
In
vivo
fluorescence
imaging
using
Cellvizio®
prototype
 A about B C In vivo biodistribution and kidney clearance of αvβ3 positron emissionMicroprobe tomography, have provided huge information positron emission tomography, have provided huge information about UltraMiniO Using
the
endoscopic
system,
we
demonstrated
that
we
can
simultaneously
observe
th Aner
 fDOT
 imaging
 ,
 1mg
 the of
structure FITC‐dextran
 (500
 kDa)
 the structure and function of the brain during the last years but the low integrin molecular marker and function of the brainwas
 during the last years but the low Resolution 1,4 µm resolution and acquisition times limits the information that can be intravenously
 injected
 
 in
 resolution animals
 before
 surgery.
 Then,
the information that can be blood
 vessels.
 We
 observed
 a
 high
 accumula^on
 of
 Angiostamp®
 surounding
 blood
 v and acquisition times
 limits Working 60 µm obtained with Distance these techniques. obtained withwas
 theseperformed
 techniques. with
 the
 Animal model fiber-optics bundle Fluorescence
 imaging
 at
 the
 cellular
 level
 A 30000 new technology developed by MaunaKea®, called Fibered Angiostamp®
was
localised
close
to
blood
vessels
of
healthy
^ssue
such
as
muscle,
s A new technology developed by MaunaKea®, called Fibered Fluorescence Microscopy, is trying to fill the gap between the existing Female nude mouse bearing MDA MB231 tumor fibered
confocal
microscope
Cellvizio®
Dual
Band
from
 
Mauna
Kea
 brain imaging techniques. The Cellvizio microscope, based in a fiber Cellvizio®
allows
us
to
confirm
that
the
macroscopic
image
obtain
by
fDOT
correspon Fluorescence Microscopy, is trying to fill the gap between the existing 50 µm 50 µm 50 µm xenograft underwent intravenous injection of 1 mg optic probe that transport the emission and fluorescent light to the Technologies.
 The
 device
 consists
 in
 a
 flexible
 sub‐millimetric
 brain imaging techniques. The Cellvizio microscope, based in a fiber Hindlimb Hindlimb Tumor FITC-Dextran 500 kDa (Sigma-Aldrich) andthe
 10 nmol maybe
 also
 to
 uptake
 by
 tumor
 associated
 macrophages
 expressing
 αvβ3
 Integ scanning unit, is able to acquire confocal images with cellular microprobe
 containing
 thousands
 op^cal
 that
 light
and fluorescent light to the opticof
probe thatfibers
 transport thecarry
 emission Redox imaging. Lentiviral vector targeting redox sensitive protein (roGFP) to Angiostamp® 700 (Raft RGD, fluOptics) resolution (3 μm axial resolution) of deep brain regions in live animals. monitoring
 of
 two
 fluorescent
 signals
 by
 endomicroscopy
 can
 be
 useful
 to
 v the mitochondria was designed and produced. Image acquisition revealed scanning unit, is and
 able660
 to nm
 acquire confocal images with cellular However, it is necessary to introduce a fluorescent dye or protein to from
 two
 con^nuous
 laser
 source
 at
 488
 nm
 to
 the
 redox to spikes of 660 Individual 488 nm(roGFP) nm cells in the hippocampus Mergeof live mice. ImageCell® Redox the imaging. Lentiviral vector targeting redox sensitive protein visualize cells, which usually generates background during the resolution (3 μm axial resolution) of deep brain regions in live animals. D macroscopic
imaging
and
it
opens
a
new
avenue
to
monitor
in
vivo
molecular
events
 software was used to record images at a frequency of 12 Hz, for quantification living
 ^ssue.
 The
 fluorescence
 emioed
 aner
 excita^on
 by
 the
 imaging process. Optimization of viral vector technology accordingly the mitochondria was designed and produced. Image acquisition revealed However, it is necessary to introduce a fluorescent dye or protein to and representation of the intensity of the fluorescent signal. Raw data of A, B | Optical biopsy of hindlimb vessels. with the characteristics of the technique can improve the quality of the redox spikes of Individual cells in the hippocampus of live mice. ImageCell® fluorophores
 staining
 the
 ^ssue
 species
 is
 sent
 back
 to
 the
 signal intensity was plotted for every time point. visualize the cells, which usually generates background during the images acquired Endothelial wall cells visible as well as blood flow software was with usedthis to microscope. record images at a frequency of 12 Hz, for quantification apparatus,
where
a
dedicated
set
of
algorithms
reconstructs
images
 imaging process. Optimization of viral vector technology accordingly and representation of the intensity of the fluorescent signal. Raw data of C | Tumor vessels and tumor associated with the characteristics of the technique can improve the quality of the in
real
^me
at
a
frame
rate
of
12
frames
per
second.
The
probe
that
 signal intensity was plotted for every time point. macrophages mixed with endothelial cells FIBERED FLUORESCENCE MICROSCOPY CALCIUM IMAGING OF SEVERAL CELLS IN LIVE MICE images acquired with this microscope. was
 used
 is
 a
 UltraMiniO
 probe
 with
 30,000
 op^cal
 fibers,
 a
 Cellvizio ® Dual Band is a confocal microscope which makes D | Optical slicing of the kidney, exhibiting [1] Garanger, E., Boturyn, D., Jin, Z., Dumy,50P.,µm Favrot, M.C. and Coll, J.L. (2005) The
authors
would
like
to
th use of a 488 or 660 nm excitation which is injected one by AngioStamp® elimination beside vessels. GCAMP3 IMAGING 50 µm 50 µm IN THE HIPPOCAMPUS 240x240
µm
field
of
view,
and
a
1.4
µm
lateral
resolu^on.

IN VIVO OUTCOMES

MOLECULAR VIROLOGY & GENE THERAPY LEUVEN VIRAL VECTOR CORE - LVVC

NEUROBIOLOGY & GENE THERAPY

MOLECULAR VIROLOGY & GENE THERAPY LEUVEN VIRAL VECTOR CORE - LVVC

NEUROBIOLOGY & GENE THERAPY

Conclusions

PRINCIPLE & ARCHITECTURE

Literature cited

Acknowled

New multifunctional molecular conjugate vector for targeting, imaging, and assistance
 for
 fDOT/CT
 ima one in tens of thousands of tiny fibers optics grouped in a Kidney vasculature AngioStamp® clearance Overlay of the two FIBERED FLUORESCENCE MICROSCOPY CALCIUM IMAGING OF SEVERAL CELLS FITC IN LIVE MICEtherapy tumors. Mol Ther, channels 12, 1168-1175. “Agence
Na^onale
pour
la
R Dextran in theof glomerulus Fiber bundle flexible fiber bundle. Excitation is conducted by the fibers European
 Molecular
 Ima down to the tissue to be examined whereGCAMP3 it is focused by Dynamic mouse colon imaging and macrophages IMAGING IN THE HIPPOCAMPUS [2] Garofalakis, A., Dubois, A., Kuhnast, B., Dupont, D.M., Janssens, I., LSH‐2004‐503569].

 Microscope description. system is composed of 2 main parts: laser some distal optics which defines the field of view,The the lateral 488 nm 660 nm Merge Scheme
2:
IllustraEon
of
different
part
of
the
tumor
that
can
be targetting during Mackiewicz, N., Dolle, F., Tavitian, B. and Duconge, F. (2010) In inflammation vivo scanning unit and the fiber optic probe. The light from a photodiode and the axial resolution of the system. Endogenous or 
imaged
by
the
Cellvizio®
Dual
Band laser is injected in every microfiber optic
 of the probe, which transports validation of free-space fluorescence tomography using nuclear imaging. Opt exogenous fluorescence is thenthe produced, Animal model light till thewhich tissue. is Thecollected emitted light is transported by the same 35, of3024-3026. CalciumLett, Imaging several cells in the hippocampus of live mice. Calcium microfiber the detector. towards The S-300 probe used for these experiments by the very same individual fiber and tillredirected a Balb/c mouse colon inflammation model sensitive protein GCaMP3 was expressed employing AAV vectors. ImageCell® contains 10.000 microfibers. software was used to record images at a frequency of 40 Hz, Topical spray of Acryflavine to reveal cypts singleof detector, an laser avalanche photodiode (APD). Scanning Microscope description. The system is composed 2 main parts: Procedure. Animals were anaesthetized by intra-peritoneal injection of the laser the proximal ketamine/medetomidine end of the fiber and bundle scanning unit and the fiber optic probe. The light fromonto a photodiode placed inisa stereotactic device. The probe GCAMP3 IMAGING IN THE OLFACTORY BULBstructure Please
contact:
francois 0,5 mg AminoSPARK (Perkin Elmer) nIR was slowly introduced in the target brain area and images acquired at laser is injected in every microfiber optic of the probe, which performed by atransports combination of a two fast oscillating mirrors, 50 µm 50 µm 50 µm fluorescent nanoparticle intravenous 



























or
frede the light till the tissue. The emitted light is providing transported the same anby overall frame rate 12Hz of 9 frequency, to 50 frames per second Image processing. ImageCell® to select regions of mice. Calcium Calcium Imaging of severalsoftware cells inwas theused hippocampus of live Distal optics administration (tail vein) microfiber till the detector. The S-300 probe used for these experiments Colon crypts Macrophages Macrophages (fps), which compensates for interest, motion artifacts. to protein quantify the Dedicated intensity of the fluorescent signal and to sensitive GCaMP3 was expressed employing AAV vectors. ImageCell® Real Time Acryflavine AminoSPARK 680 distribution contains 10.000 microfibers. Vessels are visible by negative contrast represent the data. Raw datatoof images signal intensity was plotted forHz, every image processing then operates in real time first software was used to record at a frequency of 40 Image Processing time point. Procedure. Animals were anaesthetized by intra-peritoneal injection of compensate for fiber-to-fiber differences in transmission and Tissue ketamine/medetomidine and placed in a stereotactic device. The probe GCAMP3 IMAGING IN THE OLFACTORY BULB Scheme
1:
Cellvizio®
Dual
Band
system
 background, but also to remove the well-known INCREASED SIGNAL/NOISEfiber RATIO AFTER OPTIMIZATION was slowly introduced in the target brain area and images acquired at In vivo quantification of Calcium spikes in honeycomb pattern and reconstruct a smooth and readable 12Hz frequency, olfactory bulb neurons using GCaMP3 Image processing. ImageCell® software wasimage. used to select regions of

Confocal microscope

For further

interest, to quantify the intensity of the fluorescent signal and to represent the data. Raw data of signal intensity was plotted for every time point.

INCREASED SIGNAL/NOISE RATIO AFTER OPTIMIZATION

Animal model Balb/c mouse, GCaMP3 loaded AAV local transfection. HIPPOCAMPUS HIGH TITERS HIPPOCAMPUS LOW TITERS HIPPOCAMPUS OPTIMIZED A 300 µm bevelled probe is inserted into the Conventional and optimized viral vectors were stereotactically injected with In vivo neural activation Calcium spikes Calcium Imaging of Quantification several cells inofthe olfactory bulb of live mice. Calcium bulb under a stereotaxic frame viral vectors engineered to express GFP in the hippocampus. Comparison olfactory in the olfactory bulb into Regions of interest sensitive protein GCaMP3 was expressed employing AAV vectors. Sequential of the signal/noise ratio after conventional (high and low titers) and optimized viral vectors transduction was carried out.

activation of neighboring cells was plotted at frequency of 12Hz.

C O N C L U S I O N S • Optimized viral vector technology increased the signal/noise ratio of Fibered Fluorescence Microscopy images in the hippocampus of live mice.

References

• GCaMP3 allows to record calcium levels of several cells in live mice using this new technique. HIPPOCAMPUS HIGH TITERS

HIPPOCAMPUS LOW TITERS

HIPPOCAMPUS OPTIMIZED

Conventional and optimized viral vectors were stereotactically injected with viral vectors engineered to express GFP in the hippocampus. Comparison of the signal/noise ratio after conventional (high and low titers) and optimized viral vectors transduction was carried out.

• Mitochondrial redox state can be monitored in vivo using roGFP in vivo with cellular resolution.

1- Vercauteren et al., Multicolor pCLE, SPIE Bios 2013, Calcium Imaging of several cells in the olfactory bulb of live mice. Calcium sensitive protein GCaMP3 waswasexpressed employing AAV vectors. Sequential ACKNOWLEDGEMENTS. A plasmid for mito-roGFP provided by S.J. Remington (University of2Oregon,USA). GCaMP3 from Addgene (L. Looger).of This work has been supported by IWT-SBO/060838 Brainstim, Brazo et was al.,obtained Improvement Fibered Fluorescence Microscopy SCIL programme financing PF/10/019 and IWT-O&O JANSSEN-DEPVEGF projects. activation of neighboring cells was plotted at frequency of 12Hz.

C O N C L U S I O N S

images of

individual cells in the brain of live mice, WMIC 2012 3- Ducongé et al, Simultaneous imaging of two different signals using a new fibered confocal microscopy system, WMIC 2011

• Optimized viral vector technology increased the signal/noise ratio of Fibered Fluorescence Microscopy images in the hippocampus of live mice. • GCaMP3 allows to record calcium levels of several cells in live mice using this new technique. • Mitochondrial redox state can be monitored in vivo using roGFP in vivo with cellular resolution.

ACKNOWLEDGEMENTS. A plasmid for mito-roGFP was provided by S.J. Remington (University of Oregon,USA). GCaMP3 was obtained from Addgene (L. Looger). This work has been supported by IWT-SBO/060838 Brainstim, SCIL programme financing PF/10/019 and IWT-O&O JANSSEN-DEPVEGF projects.

t c a t Con ! us www.cellviziolab.com cellbiziolab@maunakeatech.com


Poster EMIM 2013