Laser-assisted adsorption by photobleaching.
This chapter presents a simple method to produce substrate-bound protein patterns of micrometer resolution. Our approach uses only low power visible lasers and commercially available reagents to obtain arbitrary patterns of wide concentration range. We provide useful and detailed information on how to assemble the experimental setup to create engineered cell culture substrates using laser scanning or widefield illumination modalities. A protocol that includes the biochemistry, the optics, and the computer programming needed to fabricate functional micropatterns of single and multiple components is explained for readers without experience in optical engineering. Finally, we introduce a novel widefield illumination scheme for fabricating large surface patterns as well as how to make simple patterns using a standard commercial confocal microscope.
Centre de Recherche de l'Hôpital Maisonneuve-Rosemont, Montréal, Québec, Canada; Département d'Ophtalmologie et Institut de Génie Biomédical, Université de Montréal, Montréal, Québec, Canada.
Jonathan M
JM
Bélisle
Centre de Recherche de l'Hôpital Maisonneuve-Rosemont, Montréal, Québec, Canada.
Javier
J
Mazzaferri
Centre de Recherche de l'Hôpital Maisonneuve-Rosemont, Montréal, Québec, Canada; Département d'Ophtalmologie et Institut de Génie Biomédical, Université de Montréal, Montréal, Québec, Canada.
Santiago
S
Costantino
D000327:Adsorption; D018929:Cell Culture Techniques / Q000379:methods*; D007091:Image Processing, Computer-Assisted / Q000379:methods; D007834:Lasers; D008027:Light; D018613:Microscopy, Confocal / Q000379:methods; D055095:Optics and Photonics; D038761:Photobleaching
Axonal guidance; LAPAP; Laser micro-engineering; Photobleaching; Protein gradient; Protein patterning; Substrate engineering; Substrate protein patterning