title: Multi-sample Arabidopsis Growth and Imaging Chamber (MAGIC) for long term imaging in the ZEISS Lightsheet Z.1.
abstract: Time-course imaging experiments on live organisms are critical for understanding the dynamics of growth and development. Light-sheet microscopy has advanced the field of long-term imaging of live specimens by significantly reducing photo-toxicity and allowing fast acquisition of three-dimensional data over time. However, current light-sheet technology does not allow the imaging of multiple plant specimens in parallel. To achieve higher throughput, we have developed a Multi-sample Arabidopsis Growth and Imaging Chamber (MAGIC) that provides near-physiological imaging conditions and allows high-throughput time-course imaging experiments in the ZEISS Lightsheet Z.1. Here, we illustrate MAGIC's imaging capabilities by following cell divisions, as an indicator of plant growth and development, over prolonged time periods. To automatically quantify the number of cell divisions in long-term experiments, we present a FIJI-based image processing pipeline. We demonstrate that plants imaged with our chamber undergo cell divisions for >16 times longer than those with the glass capillary system supplied by the ZEISS Z1.
authors.affiliation: Plant and Microbial Biology Department, North Carolina State University, Raleigh, USA.
authors.forename: Maria Angels
authors.initials: MA
authors.lastname: de Luis Balaguer
authors.affiliation: Plant and Microbial Biology Department, North Carolina State University, Raleigh, USA.
authors.forename: Marina
authors.initials: M
authors.lastname: Ramos-Pezzotti
authors.affiliation: Plant and Microbial Biology Department, North Carolina State University, Raleigh, USA; Biological Engineering Department, North Carolina State University, Raleigh, USA.
authors.forename: Morjan B
authors.initials: MB
authors.lastname: Rahhal
authors.affiliation: Plant and Microbial Biology Department, North Carolina State University, Raleigh, USA; Physics and Biology Departments, North Carolina State University, Raleigh, USA; Industrial and Systems Engineering Department, North Carolina State University, Raleigh, USA.
authors.forename: Charles E
authors.initials: CE
authors.lastname: Melvin
authors.affiliation: Cellular and Molecular Imaging Facility, North Carolina State University, Raleigh, USA.
authors.forename: Eva
authors.initials: E
authors.lastname: Johannes
authors.affiliation: Industrial and Systems Engineering Department, North Carolina State University, Raleigh, USA. Electronic address: tjhorn.ims@gmail.com.
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Horn
authors.affiliation: Plant and Microbial Biology Department, North Carolina State University, Raleigh, USA. Electronic address: ross_sozzani@ncsu.edu.
authors.forename: Rosangela
authors.initials: R
authors.lastname: Sozzani
mesh_terms: D017360:Arabidopsis / Q000648:ultrastructure*; D029681:Arabidopsis Proteins / Q000032:analysis; D002455:Cell Division; D017076:Computer-Aided Design; D019926:Cyclin B / Q000032:analysis; D004867:Equipment Design; D049452:Green Fluorescent Proteins / Q000032:analysis; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000295:instrumentation* / Q000379:methods; D018517:Plant Roots / Q000648:ultrastructure; D066330:Printing, Three-Dimensional; D011993:Recombinant Fusion Proteins / Q000032:analysis; D059008:Time-Lapse Imaging / Q000295:instrumentation* / Q000379:methods
chemical_list: D029681:Arabidopsis Proteins; C462528:CycB1 protein, Arabidopsis; D019926:Cyclin B; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins
keywords: 3D printing; Arabidopsis root; Light sheet; Time-lapse imaging