TY - JOUR
T1 - Light-Induced Molecular Adsorption of Proteins Using the PRIMO System for Micro-Patterning to Study Cell Responses to Extracellular Matrix Proteins
AU - Melero, Cristina
AU - Kolmogorova, Aljona
AU - Atherton, Paul
AU - Derby, Brian
AU - Reid, Adam
AU - Jansen, Karin
AU - Ballestrem, Christoph
PY - 2019/10/11
Y1 - 2019/10/11
N2 - Cells sense a variety of extracellular cues, including the composition and geometry of the extracellular matrix, which is synthesized and remodeled by the cells themselves. Here, we present the method of Light-Induced Molecular Adsorption of Proteins (LIMAP) using the PRIMO system as a patterning technique to produce micro-patterned extracellular matrix (ECM) substrates using a single or combination of proteins. The method enables printing of ECM patterns in micron resolution with excellent reproducibility. We provide a step-by-step protocol and demonstrate how this can be applied to study the processes of neuronal pathfinding. LIMAP has significant advantages over existing micro-printing methods in terms of the ease of patterning more than one component and the ability to generate a pattern with any geometry or gradient. The protocol can easily be adapted to study the contribution of almost any chemical component towards cell fate and cell behavior. Finally, we discuss common issues that can arise and how these can be avoided.
AB - Cells sense a variety of extracellular cues, including the composition and geometry of the extracellular matrix, which is synthesized and remodeled by the cells themselves. Here, we present the method of Light-Induced Molecular Adsorption of Proteins (LIMAP) using the PRIMO system as a patterning technique to produce micro-patterned extracellular matrix (ECM) substrates using a single or combination of proteins. The method enables printing of ECM patterns in micron resolution with excellent reproducibility. We provide a step-by-step protocol and demonstrate how this can be applied to study the processes of neuronal pathfinding. LIMAP has significant advantages over existing micro-printing methods in terms of the ease of patterning more than one component and the ability to generate a pattern with any geometry or gradient. The protocol can easily be adapted to study the contribution of almost any chemical component towards cell fate and cell behavior. Finally, we discuss common issues that can arise and how these can be avoided.
KW - Adsorption
KW - Extracellular Matrix/chemistry
KW - Extracellular Matrix Proteins/chemistry
KW - Lasers
KW - Micro-Electrical-Mechanical Systems/instrumentation
KW - Printing, Three-Dimensional/instrumentation
KW - Reproducibility of Results
U2 - 10.3791/60092
DO - 10.3791/60092
M3 - Article
C2 - 31657796
SN - 1940-087X
JO - Journal of visualized experiments : JoVE
JF - Journal of visualized experiments : JoVE
IS - 152
ER -