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Problem to solve:

Coupling gene expression dynamics and chemical signalling to within-organ growth patterns from the macroscopic to the cellular scale remains a major challenge in plant biology. Indeed, entire seedlings cannot properly be imaged using conventional microscope configurations.

THE VIP-box

Disadvantages of the current imaging methods

Overview-of-the-vertical-microscope-setup-The-vertically-mounted-confocal-microscope_Q320.

Von Wangenheim et al., eLife 2017

Conventional imaging of plants induces many stresses:

Mecanical stresses due to the plant imaged between a slide and a coverslip.

Stresses due to gravitropism because imaged in horizontal position.

Phototoxicity induced by the power of the laser.

Phototropism periods not respected in long time experiments.

Confocal microscope setups for vertical imaging of the plants:

Require high skilled capabilities.

Only few home-made setups in the world..

Expensive.

The VIP-box (Vertical Imaging of the Plants) is an open-source homemade and low-cost imaging plateform in which seedlings can be vertically grown and imaged over a long-time period and at high-throughput.

Design of the VIP-box

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The VIP-box consists in a 3D-printed system made of 6 connected pieces. The system has been designed to be pluged on an Axiozoom microscope but can be easily adapted to other systems.
Seedlings are growing on an agarose round petri dishe which is plugged in the seedling holder. Doors are used to keep the seedling in the dark as much as possible to avoid phototropism issues.
Since the medium is drying quickly, a channel for water around the seedling holder has been incorporated to the system.

The VIP-box in action

APPLICATIONS

Developmental biology

The VIP-box makes possible the analysis of the apical hook opening

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The spatio-temporal resolution provided is sufficient for nucleus tracking even in small plant organs like root hairs (left) and the hypocotyl (right)

Biosensors imaging

pH imaging using the Apo-pHusion biosensor (Gettings et al., 2012).

Basic pH: red

Acidic pH: blue

@2023 Created by Fabien MIART. All of the images have been acquired in collaboration with IJPB, INRAE, CRRBM and ICAP, UPJV.

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