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Two‐photon fluorescent microlithography for live‐cell imaging
Author(s) -
Costantino Santiago,
Heinze Katrin G.,
Martínez Oscar E.,
De Koninck Paul,
Wiseman Paul W.
Publication year - 2005
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.20247
Subject(s) - microcontact printing , fluorescence , lithography , materials science , fluorescence microscope , microscopy , soft lithography , nanotechnology , live cell imaging , etching (microfabrication) , optics , fabrication , optoelectronics , chemistry , cell , biochemistry , physics , medicine , alternative medicine , pathology , layer (electronics)
Fluorescent dyes added to UV‐cure resins allow the rapid fabrication of fluorescent micropatterns on standard glass coverslips by two‐photon optical lithography. We use this lithographic method to tailor fiduciary markers, focal references, and calibration tools, for fluorescence and laser scanning microscopy. Fluorescent microlithography provides spatial landmarks to quantify molecular transport, cell growth and migration, and to compensate for focal drift during time‐lapse imaging. We show that the fluorescent patterned microstructures are biocompatible with cultures of mammalian cell lines and hippocampal neurons. Furthermore, the high‐relief topology of the lithographed substrates is utilized as a mold for poly(dimethylsiloxane) stamps to create protein patterns by microcontact printing, representing an alternative to the current etching techniques. We present two different applications of such protein patterns for localizing cell adhesion and guidance of neurite outgrowth. Microsc. Res. Tech. 68:272–276, 2005. © 2005 Wiley‐Liss, Inc.