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Rapid prototyping of 3D micro- nanostructures to explore cell behaviour 4M Conference 3.10.2007 Borovets
- Introduction (IBMT)
- State of the art
- Two Photon polymerisation (2PP)
- Experimental Setup
- Material/Biological Results
Rapid prototyping of 3D micro- nanostructures to explore cell - - PowerPoint PPT Presentation
Rapid prototyping of 3D micro- nanostructures to explore cell - - PowerPoint PPT Presentation
Rapid prototyping of 3D micro- nanostructures to explore cell behavior Herbert Schuck (F. Bauerfeld, D. Sauer, R. Le Harzic, E. Lai, T. Velten, I. Riemann, K. Knig) October 3, 2007 4M Conference Rapid prototyping of 3D micro- nanostructures
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Rapid prototyping of 3D micro- nanostructures to explore cell behaviour 4M Conference 3.10.2007 Borovets
Fraunhofer Institute for Biomedical Engineering (FhG-IBMT)
- St. Ingbert
Sulzbach Shenzhen, China Magdeburg Halle Dortmund Schmallenberg Aachen Euskirchen Darmstadt Jena Chemnitz Dresden Itzehoe Rostock Bremen Hanover Brunswick Kaiserslautern Freiburg Würzburg Nuremberg Holzkirchen Karlsruhe Duisburg Cottbus Ilmenau Erlangen Oberpfaffenhofen Pfinztal Munich Freising Wertheim Oberhausen Paderborn Frankfurt Garmisch- Efringen-Kirchen Partenkirchen Stuttgart Berlin Teltow
- St. Augustin
Saarbrücken
- St. Ingbert
Potsdam Golm Lübeck
- Interface between technology and biology (medicine)
Potzdam-Golm Lübeck
- Microsystems
- Laser Medicine
- NMR
- Ultrasound
- Telemedicine Telematics
- Neuroprostetics
- Cryobiophysics
- Biohybrid System
- Cell Differentiation
- Cell Technology
- Cellular
Biotechnology
- Biochips
- Molecular
Bioanalytics
- Bioelectronics
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Rapid prototyping of 3D micro- nanostructures to explore cell behaviour 4M Conference 3.10.2007 Borovets
State of the art
Technology surface properties:
~ 100 x 100 nm
SEM of a human fibroblast cell
- n structured Si surface
Intel P4 Processor built on 90 nm technology Electron micrograph
- f an influenza virus
Biology
- adhesion
- attachment
- molecular surface interactions
- morphology
- rientation
- topography
} }
~ 100 nm ~ 20 µm
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Rapid prototyping of 3D micro- nanostructures to explore cell behaviour 4M Conference 3.10.2007 Borovets
Biological problems/ requirements
- cell growth
- cell differentiation
- similarity to in vivo conditions
- culture container
- bioreactors
- with reproducible in vivo surface conditions
}
}
- matrices
. Problem: many different cell types requires different environment conditions surface production method
- investigate cells
- produce tissue
- develop therapeutic
agents .
- flexible change of geometry (real 3D)
- rapid
- considering the different biological conditions
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Rapid prototyping of 3D micro- nanostructures to explore cell behaviour 4M Conference 3.10.2007 Borovets
Two Photon Polymerisation (2PP)
One photon excitation e.g. 365 nm Two photon excitation (TPE) e.g. 730 nm Photoresist focal volume
S0 S1 E1 S0 S1 ½E1 ½E1
h h h h ν ν ν ν365nm
365nm 365nm 365nm = 2
= 2 = 2 = 2 h ν ν ν ν730nm
- t
t t t abs
abs abs abs ≤
≤ ≤ ≤ 10 10 10 10 -
- 15
15 15 15 s
s s s photon flux density ≥ 1024 [photons cm-2 s-1] fs laser pulses (e.g. Ti:Sa) Photoresist: polymerised ellipsoid volume in space
- abs. at 365 nm
- trans. at 730 nm
TPE volume Laserbeam lens φxy ~ 100 nm
real 3D structures with undercuts CAM
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Rapid prototyping of 3D micro- nanostructures to explore cell behaviour 4M Conference 3.10.2007 Borovets
non amplified system Ti:Sa
(720 - 930 nm/ 90 MHz/ 150 fs/ 1,6 Wavg / Chameleon/ Coherent )
Zeiss LSM 510 Meta NLO
- Processing of photo resists with this “cheap” experimental setup
Experimental setup
Coherent Chameleon AOTF Scanning Mirrors Scanning Optics Specimen Objective Lens Dichroic Beamsplitter (DBS) DBS DBS Grating META Detector (linear PMT Array) PMT PMT Emission Filters Coherent Chameleon AOTF Scanning Mirrors Scanning Optics Specimen Objective Lens Dichroic Beamsplitter (DBS) DBS DBS Grating META Detector (linear PMT Array) PMT PMT Emission Filters
high NA objectives (1,3 oil/ 40x) Si-wafer Photo resist Cover slip
- il
2PP volume
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Rapid prototyping of 3D micro- nanostructures to explore cell behaviour 4M Conference 3.10.2007 Borovets
3D-Structuring by maskless nano lithography (50µm high)
6 µm
SU8 (negative resist)
Material results
300 nm Aspect ratio 150:1
3D cell container (FIB cut & SEM by IZM Berlin) AZ 9260
(positive resist)
10 µm
Exposure time depends on complexity
- f structure, resist, equipment, laser…..
(e.g. 250x250x50µm ~ 5min)
SU8 with fluorescent marker Problem: non destructive analysis ? 3D cell container (SU8)
(with undercuts)
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Rapid prototyping of 3D micro- nanostructures to explore cell behaviour 4M Conference 3.10.2007 Borovets
Biological results
test pattern (SU8) on silicon Spiraled test pattern (SU8 on glass)
5 µm 100 µm
(CHO [Chinese Hamster Ovary] 48h)
r ~ 480 µm
(CHO (96h)) (DPSC)
[DentalPulpaSteamCells]/96h
Concentric circles Spirals
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Rapid prototyping of 3D micro- nanostructures to explore cell behaviour 4M Conference 3.10.2007 Borovets
Summary/ Outlook
- Alignment depends on structure and cell species
- Structures with positive and negative resist
- High aspect ratio150:1
- Structures down to 100 nm
- Structures with undercuts
- Electroplating/ hot embossing experiments
- Rapid production of 3D prototype structures
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