1 research outputs found
Efficient Biofunctionalization of Polysilicon Barcodes for Adhesion to the Zona Pellucida of Mouse Embryos
Cell tracking is an emergent area in nanobiotechnology,
promising
the study of individual cells or the identification of populations
of cultured cells. In our approach, microtools designed for extracellular
tagging are prepared, because using biofunctionalized polysilicon
barcodes to tag cell membranes externally avoids the inconveniences
of cell internalization. The crucial covalent biofunctionalization
process determining the ultimate functionality was studied in order
to find the optimum conditions to link a biomolecule to a polysilicon
barcode surface using a self-assembled monolayer (SAM) as the connector.
Specifically, a lectin (wheat germ agglutinin, WGA) was used because
of its capacity to recognize some specific carbohydrates present on
the surface of most mammalian cells. Self-assembled monolayers were
prepared on polysilicon surfaces including aldehyde groups as terminal
functions to study the suitability of their covalent chemical bonding
to WGA. Some parameters, such as the polysilicon surface roughness
or the concentration of WGA, proved to be crucial for successful biofunctionalization
and bioactivity. The SAMs were characterized by contact angle measurements,
time-of-flight secondary ion mass spectrometry (TOF-SIMS), laser desorption/ionization
time-of-flight mass spectrometry (LDI-TOF MS), and atomic force microscopy
(AFM). The biofunctionalization step was also characterized by fluorescence
microscopy and, in the case of barcodes, by adhesion experiments to
the zona pellucida of mouse embryos. These experiments showed high
barcode retention rates after 96 h of culture as well as high embryo
viability to the blastocyst stage, indicating the robustness of the
biofunctionalization and, therefore, the potential of these new microtools
to be used for cell tagging