Interfacing insect neuronal neutworks with microelectronic devices [Elektronische Ressource] / vorgelegt von Anna Magdalena Reska

Areska

Sujets

• Biologie

Informations

• Publié par: rheinisch-westfalischen_technischen_hochschule_-rwth-_aachen
• Publié le: 01 janvier 2009
• Nombre de lectures: 36
• Langue: Deutsch
• Poids de l'ouvrage: 7 Mo

Extrait

Interfacing insect neuronal networks
with microelectronic devices


Von der Fakultät für Mathematik, Informatik undu rNwatissenschaften der
RWTH Aachen University zur Erlangung des akademisecnh Grades einer
Doktorin der Naturwissenschaften
vorgelegt von

Diplom-Biologin

Anna Magdalena Reska
aus Pyskowice, Polen

Berichter: Universitätsprofessor Andreas Offenheäur ss
Universitätsprofessor Peter Bräunig

Tag der mündlichen Prüfung: 16.Juli 2009

Diese Dissertation ist auf den Internetseiteonc hdsechr ulHbibliothek online verfügbar. Interfacing insect neuronal networks
with microelectronic devices
Anna Magdalena ReskaBerichte des Forschungszentrums Jülich; 4312
ISSN 0944-2952
Institute of Bio- and Nanosystems (IBN)
Bioelectronics (IBN-2) Jül-4312
D 82 (Diss., RWTH Aachen, Univ., 2009)
Vollständig frei verfügbar im Internet auf dem Jülicher Open Access Server (JUWEL)
unter http://www.fz-juelich.de/zb/juwel
Zu beziehen durch: Forschungszentrum Jülich GmbH · Zentralbibliothek, Verlag
D-52425 Jülich · Bundesrepublik Deutschland
02461 61-5220 · Telefax: 02461 61-6103 · e-mail: [e-mail] of conten ts

1 Motivation............................................................1.. .......................
2 Introduction.........................................................5. ..........................
2.1.1 Invertebrate neuronal networks.......................................................................6
2.1.2 Cricket cercal sensory system........................................................................... ..........................................7
2.1.3 Morphology of invertebrate neurionn vsi vo and in vitro..............................10. ....................
2.1.4 Primary cell culture...................................................11. .......................
2.1.5 Neurogenesis: adhesion molecules and neuicro tfracoptohrs...................................1.1 ...............
2.1.6 Network development..............................................3. ...............................1
2.1.7 Cellular patterning......................................................15. ......................
2.1.8 Outline......................................................................19. ...........
3 Materials and Methods..................................................................................21
3.1 Primary neuronal cell culture of crickets .n.e.u.r.o.n............................ ..........................21
3.1.1 Animals.............................................................. ......2.1..................
3.1.2 Procedure of cell culture..................................................................................... .......................................22
3.2 Surface coatings for primary cell culture..................................... ...........................23
3.2. 1 Uniformly conA-coated surfaces.......................................... ...........................23
3.2. 2 Surface coatings with star PEG / conA grid pa.tt.e.r.n................................... ...................2.4............
3.3 Microcontact Printing...................................................... ..2.6........................
3.4 Parylene /star PEG patterning on microelectrovniice sd efor extracellular recordings.. .....28
3.4. 1 Microelectronic devices............................................... ..............................28
3.4. 2 Parylene stencil production................................................................................ ......................................29
3.4. 3 Plasma assisted removal of star PEG with paryleennec ilsst...................... ....2.9..................
3.5 Surface characterization.................................................... .3.1..........................
3.5. 1 Ellipsometry............................................................................................................... ........3..1...........................
3.5. 2 Atomic force microscopy.............................................. .............................32
3.5. 3 Scanning electron microscope ......................................... ............................35
SEM measurements........................................................ ...3.5.......................
i Table of conten ts
Cell Fixation for SEM....................................................... ....3.7....................
Critical point drying....................................................... .....3.7...................
3.6 Data evaluation of morphological type distributionnd astatistical analysis.............3. 8...............
3.7 Electrophysiology......................................................... ....3.9....................
3.7. 1 Voltage clamp experiments................................................................................. ......................................41
3.7. 2 Current clamp experiments............................................ ............................42
3.7.3 Paired patch- clamp recordings...................................................................43
4 Ultrathin coatings with reactivity change by t imeneable functionailn vitro networks
of insect neurons..........................................................................................4.5.. ............................................
5 Surface growth restriction in insect neuronal ceullture promotesin vivo-like
phenotypes......................................................6.1. ...............................
6 Inverse patterning of cell-aversive Star PEGb ufoilrd ing neuronal networks on
microelectronic devices............................................................................................................................83
7 Discussion.........................................................9.7. ..........................
8 Summary......................................................1.0.1. .............................
9 Zusammenfassung...................................................................................................................................103

ii List of illustrations
List of illustrations
2.1. Schematic drawing of cricket cercal system
2.2. Model of the cercal sensory neuronal system.
3.1. Female cricGkreytll us bimaculatus.
3.2. a) Ventral view on abdomGerynl luosf bimaculatus.
b) Isolated terminal abdominal ganglion.
3.3. Procedure of star PEG layer formation.
3.4. Cross-linking reaction of star PEG.
3.5. Creation of a master for molding PDMS mmicprso.s ta
3.6. Principle of microcontact printing (µCP).
3.7. Two types of microelectronic devices
3.8. Schematic of plasma assisted removal ofE Gs tawr iPth parylene stencils.
3.9. Laser beam polarization during ellipsomeatsruyr ement.
3.10. Schematic drawing of atomic microscope .s et-up
3.11. AFM tip scanning a surface in tapping mode.
3.12. Schematic of a scanning electron microscope.
3.13. Colourized SEM micrograph of two crickeotn sn egruorwn on star PEG/ conA patterned surface.
3.14. Three different pattern types for quainotni ficofa tcell morphology development upon adhesion
surface restriction.
3.15. Measuring circuit of patch clamp experiment.
3.16. Equivalent circuit of patch clamp expe riment.
3.17. Characteristic current trend during a vcollatmagpe experiment.
3.18. Typical voltage clamp recording of cricukroent. ne
3.19. Typical current clamp recording of a imkuinltgi csrpicket neuron
3.20. Schematic drawing of paired patch-clamp imexepnet.r
3.21. Example of a recording of two synaptincnalelyc tecod cricket neurons in current clamp mode.
4.1. Chemistry of the star PEG system and itohnes rtehaact tlead to film formation.
4.2. Confocal laser scan image of rhodamine-ereledd- lacbonA printed on a star PEG surface.
4.3. Pattern fidelity of cricket neurons gr ocwonn Ao/nst ara PEG surface after A) 8 and B) 10 d ays in
culture, visualized by SEM (A) and phase-conitgrhastt mlicroscopy (B).
iii List of illustrations
4.4. A) Phase-contrast light microscopy of doautcbhle- clapmp recording obtained from a
morphologically connected pair of two cricketn sn eafuteror 5 days in culture on a/ sctoarn A
PEG surface.
B) Stimulation of the presynaptic cell led nto paoctteiontials in this cell (black, top lined) followe
by correlated and summated postsynaptic post eint iatlhe postsynaptic cell (grey, lower line).
5.1. Three different node and line patternsd pfroorv indeeuronal growth.
5.2. Representative images of the three deifflel remnotr pchologies of cricket neurons grown on
planar conA-coated glass surfaces.
5.3. Calcein stained cricket neurons on con Ag-lacsosa tesldides (a-c) and star PEG / conA patterns
(d-f).
5.4. Comparison of percentages of each morphlo ltoygpieca on different surface pattern.
5.5. Comparison of cell morphology types depoen ndsuinrfga ce pattern.
5.6. Electrophysiological response types on luyn icfoornmA-coated (white background) and pre-
patterned (grey background) surfaces.
5.7. Resting membrane potentials of the fouorp heysleiocltorgical response types.
5.8. Maximal action potential amplitude upon 1 unrrAe nct stimulation of electrophysiological
respo