Multi-sample Arabidopsis Growth and Imaging Chamber (MAGIC) for long term imaging in the ZEISS Lightsheet Z.1

Maria Angels de Luis Balaguer; Marina Ramos-Pezzotti; Morjan B Rahhal; Charles E Melvin; Eva Johannes; Timothy J Horn; Rosangela Sozzani

doi:10.1016/j.ydbio.2016.05.029

Multi-sample Arabidopsis Growth and Imaging Chamber (MAGIC) for long term imaging in the ZEISS Lightsheet Z.1

Dev Biol. 2016 Nov 1;419(1):19-25. doi: 10.1016/j.ydbio.2016.05.029. Epub 2016 May 26.

Authors

Maria Angels de Luis Balaguer¹, Marina Ramos-Pezzotti¹, Morjan B Rahhal², Charles E Melvin³, Eva Johannes⁴, Timothy J Horn⁵, Rosangela Sozzani⁶

Affiliations

¹ Plant and Microbial Biology Department, North Carolina State University, Raleigh, USA.
² Plant and Microbial Biology Department, North Carolina State University, Raleigh, USA; Biological Engineering Department, North Carolina State University, Raleigh, USA.
³ 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.
⁴ Cellular and Molecular Imaging Facility, North Carolina State University, Raleigh, USA.
⁵ Industrial and Systems Engineering Department, North Carolina State University, Raleigh, USA. Electronic address: tjhorn.ims@gmail.com.
⁶ Plant and Microbial Biology Department, North Carolina State University, Raleigh, USA. Electronic address: ross_sozzani@ncsu.edu.

PMID: 27235815
DOI: 10.1016/j.ydbio.2016.05.029

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.

Keywords: 3D printing; Arabidopsis root; Light sheet; Time-lapse imaging.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Arabidopsis / ultrastructure*
Arabidopsis Proteins / analysis
Cell Division
Computer-Aided Design
Cyclin B / analysis
Equipment Design
Green Fluorescent Proteins / analysis
Image Processing, Computer-Assisted / methods*
Microscopy, Fluorescence / instrumentation*
Microscopy, Fluorescence / methods
Plant Roots / ultrastructure
Printing, Three-Dimensional
Recombinant Fusion Proteins / analysis
Time-Lapse Imaging / instrumentation*
Time-Lapse Imaging / methods

Substances

Arabidopsis Proteins
CycB1 protein, Arabidopsis
Cyclin B
Recombinant Fusion Proteins
Green Fluorescent Proteins

Grants and funding

MCB 1517058/Biotechnology and Biological Sciences Research Council/United Kingdom