SNOTRA LLC has developed a new scanning probe nanotomography technology that allows for non-destructive 3D analysis of native nanoscale structures in soft materials like biomaterials and polymers. This novel technology combines scanning probe microscopy and cryoultramicrotomy to provide 3D structural information not available through other existing methods. SNOTRA has built a working prototype, published results in scientific journals, and is seeking partners for commercialization and further development through grant funding.

1. SNOTRA LLC.
Scanning probe nanotomography –
a new technology for analysis and
diagnostics of biomaterials and polymers

2. The Problem We Address
 Nanoscale structure analysis of majority
(~80%) of biological and soft polymer materials
require cryosectioning and investigation at low
temperatures.
 Existing approaches (f.e. cryoTEM) are very
expensive and suffer from a number of issues (low
EM contrast, beam damage, etc), especially for 3D
analysis.
 Thus a number of problems concerning
nanoscale analysis of biological, polymer and other
soft materials still do not have an adequate solution

3. SNOTRA LLC.
The Goal:
Development and commercialization of methodology and
combined analytical system for nanotomography – non-
destructive three-dimensional analysis of native
nanoscale structures in a wide range of soft materials.
The Solution is based on combination of
scanning probe microscopy (nanoscale analysis of
surface features)
and
cryoultramicrotomy (ultrathin sectioning of soft materials
at temperatures down to -190°С).

4. Scanning probe
cryo
nanotomography
Scanning probe
microscopy
(nanoscale surface
analysis)
Cryo ultramicrotomy
(ultrathin sections - down to 10
nm – of frozen samples)
2D (XY)
1D (Z)
+
=
3D(XYZ)

5. SNOTRA LLC.
A novel technology of scanning probe nanotomography
Is applicable to: biomaterials, biopolymers, other soft or hydrated
polymers & materials (e.g. resins), nanoemulsions
Is ~ 10 times cheaper than currently available competing
methods/devices (f.e.TEM electron tomography, SEM/FIB)
Provides access to unique information about nanostructure – not
available by other existing methods.
How advantage is secured by the company
 IP protection (Russian patent filed, EU patent applied, exclusive
license agreement for EU patent application is signed),
 High-ranked and well-known scientists in the field are involved
(f.e. Dr. Prof. Ferdinand Hofer, Head of FELMI/ZFE Graz, Austria),
 Working prototype (proof-of-the-concept) of the system is
successfully built and 2 prototype systems are installed.
 Results are published in scientific journals and disseminated on
scientific conferences.

6. SNOTRA LLC.
The company
Is established by inventors personally, Federal Research Center for
Transplantology and Artificial Organs (Moscow), and Center of
Innovative Enterprise (Moscow State University)
Is cooperating with one of the world leading scientific centers in the
field of the project – Austrian Centre for Electron Microscopy
(FELMI/ZFE Graz, Austria)
Is a participant of Skolkovo Foundation

7. AFM TEM
AFM versus TEM
AFM provides
the data on
molecular
structure based
on the analysis
of the
morphology
and mecha-
nical properties
of the surface
The block sur-
face is mea-
sured directly
inside the
cryogenic
chamber – in
the environ-
ment of the
sectioning
procedure
Ultrathin
sections
( 10..90 nm )
are
transferred
into the
vacuum
chamber of
the TEM
Low contrast
on biological
and polymer
samples
Projection
image is the
average of all
the planes that
constitute the
thickness of
the section
Electron
beam
AFM probe Interacting atom
UrAc
Chemical da-
mage inside
the material

8. The methods of nanostructure analysis(biology&polymers)
Product or
Technology
Resolution
Cost
Preparation and damage
to the sample structureXY, nm Z, nm
SPM +
CryoUMT
(SNOTRA)
5..10 10-20
~250
k$
Intact native structures of soft
polymers and biomaterials are measured
(cryoultratomography and immediate
measurement)
Conventional
SPM,
Bruker, Asylum, ...
5..10
Нет !
измеряет
поверхность
~200
k$
Structures are damaged
(measurements at room temperature)
CryoSPM,
Omicron, JEOL, ... 5..10
>300
k$
Hard to exploit
(vacuum, liquid He or N2 environment –
not suited for bio/polymers)
SEM Tomography
(Focused-ion-beam
sectioning), FEI, Zeiss
10 ~10-20
>1
M$
Structures are damaged
(electron and ion beams, vacuum, metal
sputtering), no cryo 3D at the moment
CryoTEM
(electronic
tomography), 5
5
Sample
thickness
>
1M$
Structures are damaged
(electron and ion beams, vacuum),
projection imaging, low contrast at
No!
Measures
only the
surface

9. Tuning fork-based AFM probe
Cryo-AFM module is installed
directly into the cryochamber
Leica EM UC6/FC6
cryoultramicrotome performs ultrathin
sectioning of soft materials at
temperatures from -15 to -190°C.
Section thickness ranges from 10 nm
to 1 um.

10. SNOTRA LLC.
Applications and examples of results
by scanning probe nanotomography

11. Nitrile butadiene-rubber latex
(a) A topographical AFM image of
an epoxy embedded latex stripe
that was mounted in the cryo-
chamber of SNOTRA, cryo-
sectioned and immediately
scanned at −120°C.
(b) Immediately afterwards, the
same sample was warmed to
room temperature and then
examined using the same AFM
(c) A TEM image of the last thin
section of the NBR latex sample.
(d) A schematic description of the
topographical change of the
sample block phase that took
place during sectioning and the
following warming processes. The
scale bars in (a, b, c) are 200 nm,
and the topographical variations
in (a) was 27.2 nm, and in (b) was
35.5 nm.
A. E. Efimov, H. Gnaegi, R. Schaller, W. Grogger, F. Hofer and N. B. Matsko,, Soft Matter, 2012, 8, 9756, DOI:10.1039/c2sm26050f

12. 3D reconstructions of the antennal sensillas Placodea and
Coeloconica 12.5×13.0×0.7 µm3, 11 sections, section
thickness 70 nm. .
3D model of chitin organization of wasp antenna sensillas
АFМ
ТЕМ

13. 3D reconstruction of PA6/SAN polymer nanocomposite
e) АFМ- topography after cryosection at -80°C.
f) АFМ- topography after section at room temperature .
g, h) 3D reconstruction at -80°С: 7.9×6.2×0.75 µm и
2.0×2.0×0.75 µm, correspondingly, 125 nm sections.

14. 3D reconstruction: porous polyoxibutirate biodegradable
cell matrix for regenerative medicine use. 20 sections,
41.2×34.1×8.5 µm
V. N. Vasilets, I. V. Kazbanov, A. E. Efimov and V. I. Sevastianov, Vestnik transplantologii i iskusstvennyh organov, 2009, 9(2), 47 (in
Russian).

15. 3D reconstruction of ABS copolymer: 15 sections
x100 nm at room temperature. 13.5x13x1.5 µm

16. . .
3D reconstruction of conductive nanocomposites
3D-reconstruction of the conductivity of the
network of graphene flakes inside the PS/
graphene nanocomposite, 2.5x2.5x0.34 um
A. Alekseev et al, Adv. Func. Mater., 22, 1311 (2012).
3D-reconstruction of the conductivity of
the nanotube network inside the polysty-
rene/CNT composite, 1.8x1.6x0.26 um.
A. Alekseev, A. Efimov, K. Lu, J. Loos, Adv. Mater., 21, 4915 (2009)

17. AFM image and 3D-reconstruction of cholesteric
liquid crystal structure with implanted fluorescent
CdSe/ZnS quantum dots, 14 sections (/100 nm)
3D study of liquid crystal / quantum dots nanocomposite
A. Bobrovsky et al, Advanced Materials, 2012, DOI:
10.1002/adma.201202227;
K. Mochalov et al, Proc. SPIE 8475, Liquid Crystals XVI,
847514, 2012, doi:10.1117/12.929729

18. a
3D study of liquid crystal / quantum dots nanocomposite
3D-AFM reconstruction and 2D AFM image (4x4 um) of cholesteric LC structure with implanted
fluorescent CdSe/ZnS quantum dots (arrows indicate the same QD on AFM image and 3D
AFM reconstruction. We observe that implanted QD do not distort the planar LC structure
K. Mochalov, A. Efimov et al, Proc. SPIE 8767, Integrated Photonics: Materials, Devices, and Applications II,
876708 (2013); doi:10.1117/12.2017088

19. Scientific publications for our technology
1) A. E. Efimov, H. Gnaegi, R. Schaller, W. Grogger, F. Hofer and N. B. Matsko, Analysis of native structure of soft materials
by cryo scanning probe tomography, Soft Matter, 2012, 8, 9756, DOI:10.1039/c2sm26050f
2) K. E. Mochalov; A. Yu. Bobrovsky; V. A. Oleinikov; A. V. Sukhanova; A. E. Efimov; V. Shibaev; I. Nabiev, Novel
cholesteric materials doped with CdSe/ZnS quantum dots with photo- and electrotunable circularly polarized emission, Proc.
SPIE 8475, Liquid Crystals XVI, 847514 (October 11, 2012)
3) Bobrovsky, A., Mochalov, K., Oleinikov, V., Sukhanova, A., Prudnikau, A., Artemyev, M., Shibaev, V., Nabiev, I. Optically
and electrically controlled circularly polarized emission from cholesteric liquid crystal materials doped with semiconductor
quantum dots. Advanced Materials, 2012, DOI: 10.1002/adma.201202227
4) A. Alekseev, D. Chen, E. E. Tkalya, M. G. Ghislandi, Yu. Syurik, O. Ageev, J. Loos, and G. de With Local Organization of
Graphene Network Inside Graphene/Polymer Composites Adv. Funct. Mater. 2012, 22, 1311–1318
5) V.Mittal and N.B.Matsko, Tomography of the Hydrated Materials, in Analytical Imaging Techniques for Soft Matter
Characterization, Engineering Materials, Springer-Verlag Berlin Heidelberg, 2012, pp. 85-93
6) A. Efimov; H. Gnaegi; V. Sevastyanov; W. Grogger; F. Hofer; N. Matsko, Combination of a cryo-AFM with an
ultramicrotome for serial section cryo-tomography of soft materials - Proceedings 10th Multinational Congress on
Microscopy 2011 SEP 4-9, 2011; Urbino, ITALY. pp.707-708
7) N. B. Matsko, J. Wagner, A. Efimov, I. Haynl, S. Mitsche, W. Czapek, B. Matsko, W. Grogger, F. Hofer, Self-Sensing and
–Actuating Probes for Tapping Mode AFM Measurements of Soft Polymers at a Wide Range of Temperatures, Journal of
Modern Physics, 2011, 2, pp. 72-78
8) A. Alekseev, A. Efimov, K. Lu, J. Loos. Three-dimensional electrical property reconstruction of conductive
nanocomposites with nanometer resolution, Advanced Materials, Vol. 21, 48 (2009), рр. 4915 – 4919
9) A. Efimov, V. Sevastyanov, W. Grogger, F. Hofer, and N. Matsko. Integration of a cryo ultramicrotome and a specially
designed cryo AFM to study soft polymers and biological systems, MC2009, Vol. 2: Life Sciences, p. 25, Verlag der TU
Graz 2009.
10) A. E. Efimov, A. G: Tonevitsky, M. Dittrich & N. B. Matsko. Atomic force microscope (AFM) combined with the
ultramicrotome: a novel device for the serial section tomography and AFM/TEM complementary structural analysis of
biological and polymer samples. Journal of Microscopy, Vol. 226, Pt 3, June 2007, pp. 207–217

20. Revenue Generation
Company
Revenue
Model
Markets
Scientific & industry
customers specialized in:
1. Biomaterials
2. Polymers
3. Pharmaceuticals
4. Liquid crystals
5. Nanoemulsions and
nanoliquids
6. Other soft/hydrated
materials
Products:
1. CryoUMT/AFM
system
2. CryoUMT/Raman/
AFM system
(next stage of
development)
Service:
Analytical services with use of our nanotomography technology

21. World market estimation*
* According to report by Future Markets, Inc. 2011
We develop unique product valuable for many users of AFM/SEM/TEM in
rapidly growing Biomedical, Polymers and Nanomaterials market
segments
We plan to achieve 10 M$ market share in 5 years
AFM/SEM/TEM (total), M$ AFM/SEM/TEM,
Biomed+Polymers+Nanomaterials, M$
2010 328,44 97,6
2015
(prognosis)
588,69 217,39

22. Companies and institutions expressed interest in the technology
• Dow Chemicals (Dow Benelux BV, polymers )
• FELMI/ZFE Graz (Austria, microscopy center)
• Boeckeler Instruments (USA, equipment for microscopy preparation and ultramicrotomy)
• Anton Paar (Austria, analytical equipment)
• Boston Scientific (USA, drug translucent stents)
• Weizmann Institute (Israel, biomaterials)
• Carmel Olefins (Israel, polymers)
• Nike IHM (USA, multilayer polymers for the shoes)
• Cornell University (USA, nanofibers for textile)
• Shell International (Netherlands, Dr. R. Haswell, fuel membranes)
• General Motors, Rochester Lab (USA, fuel membranes)
• Xerox Inc. (USA, 3-D structure of ink and paint particles)
• Nova Nordisk (USA, polymers)
• Nazarbaev University (Kazakhstan, microscopy center)
• Institute of Biophysics, Chinese Academy of Sciences (structural cell biology)

23. SNOTRA LLC.
We are looking for partners:
Customers for our devices and analytical services
Partners for collaborative research projects
Partners for commercialization and market entry
Investors for the company and technology

24. SNOTRA LLC.
SNOTRA LLC. as a participant of Skolkovo Foundation is applying for a
grant funding.
Terms of financing of R&D projects by Skolkovo Foundation (stage 1):
Total project budget: up to 1 Million Euro for 3 years
Co-investment: 25% = 250 000 Euro
Granted financing: 750 000 Euro
SNOTRA LLC. is looking for co-investors for this grant application and may
offer a share in the company.

25. SNOTRA LLC.
Contact us:
General Director, CEO: Dr. Anton Efimov,
E-mail: antefimov@gmail.com
Phone: +7(916)1642475
Mail: SNOTRA LLC., Shcherbakovskaya str.,3,
Moscow, 105318 Russia