Two-photon fluorescent microlithography for live-cell imaging.

TitleTwo-photon fluorescent microlithography for live-cell imaging.
Publication TypeJournal Article
Year of Publication2005
AuthorsCostantino S, Heinze KG, Martínez OE, De Koninck P, Wiseman PW
JournalMicrosc Res Tech
Volume68
Issue5
Pagination272-6
Date Published2005 Dec 1
ISSN1059-910X
KeywordsAnimals, Animals, Newborn, Cells, Cultured, CHO Cells, Cricetinae, Dimethylpolysiloxanes, Fibronectins, Fluorescent Dyes, Green Fluorescent Proteins, Hippocampus, Humans, Image Processing, Computer-Assisted, Microscopy, Confocal, Microscopy, Fluorescence, Neurons, Photons, Polymers, Rats
Abstract

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.

DOI10.1002/jemt.20247
Alternate JournalMicrosc. Res. Tech.
PubMed ID16315236

Funding

Our research endeavors are made possible by the following agencies:

Canadian Institutes of Health Research - Instituts de recherche en santé du Canada Fonds de recherche du Québec – Nature et technologies (FRQNT)Fonds de la recherche en santé du Québec NARSAD Natural Sciences and Engineering Research Council of Canada (NSERC) - Conseil de recherche en sciences naturelles et en génie du Canada (CRSNG)innovation.caHuman Frontier Science Program