Independent synchronized control and visualization of interactions between living cells and organisms.
To investigate the early stages of cell-cell interactions occurring between living biological samples, imaging methods with appropriate spatiotemporal resolution are required. Among the techniques currently available, those based on optical trapping are promising. Methods to image trapped objects, however, in general suffer from a lack of three-dimensional resolution, due to technical constraints. Here, we have developed an original setup comprising two independent modules: holographic optical tweezers, which offer a versatile and precise way to move multiple objects simultaneously but independently, and a confocal microscope that provides fast three-dimensional image acquisition. The optical decoupling of these two modules through the same objective gives users the possibility to easily investigate very early steps in biological interactions. We illustrate the potential of this setup with an analysis of infection by the fungus Drechmeria coniospora of different developmental stages of Caenorhabditis elegans. This has allowed us to identify specific areas on the nematode's surface where fungal spores adhere preferentially. We also quantified this adhesion process for different mutant nematode strains, and thereby derive insights into the host factors that mediate fungal spore adhesion.
Centre d'Immunologie de Marseille-Luminy UM2, Aix Marseille Université, Marseille, France; Institut National de la Santé et de la Recherche Médicale (Inserm) U1104, Marseille, France; Centre National de la Recherche Scientifique (CNRS) UMR 7280, Marseille, France.
Vincent
V
Rouger
Centre d'Immunologie de Marseille-Luminy UM2, Aix Marseille Université, Marseille, France; Institut National de la Santé et de la Recherche Médicale (Inserm) U1104, Marseille, France; Centre National de la Recherche Scientifique (CNRS) UMR 7280, Marseille, France.
Guillaume
G
Bordet
Centre d'Immunologie de Marseille-Luminy UM2, Aix Marseille Université, Marseille, France; Institut National de la Santé et de la Recherche Médicale (Inserm) U1104, Marseille, France; Centre National de la Recherche Scientifique (CNRS) UMR 7280, Marseille, France.
Carole
C
Couillault
Aix Marseille Université, CNRS UMR 7249, Centrale Marseille, Institut Fresnel, Marseille, France.
Serge
S
Monneret
Centre d'Immunologie de Marseille-Luminy UM2, Aix Marseille Université, Marseille, France; Institut National de la Santé et de la Recherche Médicale (Inserm) U1104, Marseille, France; Centre National de la Recherche Scientifique (CNRS) UMR 7280, Marseille, France.
Sébastien
S
Mailfert
Centre d'Immunologie de Marseille-Luminy UM2, Aix Marseille Université, Marseille, France; Institut National de la Santé et de la Recherche Médicale (Inserm) U1104, Marseille, France; Centre National de la Recherche Scientifique (CNRS) UMR 7280, Marseille, France.
Jonathan J
JJ
Ewbank
Centre d'Immunologie de Marseille-Luminy UM2, Aix Marseille Université, Marseille, France; Institut National de la Santé et de la Recherche Médicale (Inserm) U1104, Marseille, France; Centre National de la Recherche Scientifique (CNRS) UMR 7280, Marseille, France. Electronic address: pujol@ciml.univ-mrs.fr.
Nathalie
N
Pujol
Centre d'Immunologie de Marseille-Luminy UM2, Aix Marseille Université, Marseille, France; Institut National de la Santé et de la Recherche Médicale (Inserm) U1104, Marseille, France; Centre National de la Recherche Scientifique (CNRS) UMR 7280, Marseille, France. Electronic address: marguet@ciml.univ-mrs.fr.
Didier
D
Marguet
D000818:Animals; D017173:Caenorhabditis elegans / Q000166:cytology / Q000254:growth & development / Q000382:microbiology*; D002450:Cell Communication; D006999:Hypocreales / Q000166:cytology* / Q000502:physiology*; D007909:Lenses; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods*; D009068:Movement; D052898:Optical Tweezers; D013172:Spores, Fungal / Q000166:cytology / Q000502:physiology