Development of correlative multimodal imaging using hard-soft-X-ray-and-uv-synchrotron-radiation for structural cell biology at the Mesoscale

Supervisor Organisation PhD Awarding Entity: phd location
Synchrotron Soleil
Paris Saclay University
Synchrotron Soleil, Saclay, France

Research Focus

Nowadays cell biology requires to correlate high resolution morphology and chemical information to apprehend the complexity of the inner cellular organisation of living systems.

In order to efficiently get insightful information to address challenging scientific questions using efficient correlative imaging approaches, careful and innovative sample preparations and validations are mandatory. In this context, the SOLEIL health and biology scientific section, hosted by a large facility, benefits from in-house pilot projects and a large network of synchrotron users and collaborators for developing a wide panel of live and cryo-correlative imaging projects. These unique beamlines portfolio allow combining biophysics and imaging methods for Correlative Light Electron and X-ray Microscopy as proposed in CLEXM doctoral network initiative.

The project will focus on tailored sample preparations and standard qualification for a large diversity of scientific topics in cell biology.

Methodology

The SOLEIL facility provides a wide range of photons energies, from IR to Hard X-ray coupled with state-of-the-art beamlines that cooperate to combine imaging and X-ray diffraction modalities. In addition, SOLEIL will host a 300kV cryo-EM early 2024. Proper biological sample (such as bacteria, eukaryotic cells to mono- or multicellular layers and tissues) cryo-preparation is critical for successful and relevant correlative bio imaging experiments.

It relies on the double objective that the biological sample layered on the sample holder (typically a cryoEM grid or a silicon nitride film for example) will be preserved in its physiologically relevant state while ensuring multimodal data acquisition. Sample thickness and distribution within thin vitreous (non-crystalline) water are critical parameters for successful CLEXM approaches. Therefore, proper evaluation of these sample characteristics is mandatory for sample preparation workflows validation using an innovative combination of techniques available at SOLEIL.

SOLEIL also hosts microfluidics assisted live imaging instruments (VUV beamline, multiphoton microscope) that will provide opportunities to develop original methods to link dynamics (live microscopy) and high spatial resolution (cryo-microscopy) information on biological systems.

Aim 1

Thick biological sample preparation for imaging (eukaryotic cells, tissues…)

Aim 2

Hard X-rays to characterize vitrification processes
(Diffraction patterns)

Aim 3

Hard X-rays fluorescence: elemental mapping & Common multi-technique grid cryo-holder design

Pictures Attached

Image description: Synchrotron Soleli

Role/Focus of PhD:

Mission

Elaborate and develop optimized sample preparation workflows and methodologies for various correlative bioimaging projects in cell biology.

Responsibilities and task

Developments of sample preparation for correlative cryo imaging for a wide range of photons energies, from Soft to Hard X-ray.

Education and Experience

The candidate should hold a M.Sc. or equivalent degree in Physics, Biology, Chemistry or a related discipline.

Required basic knowledge and skills

knowledge of microscopy, cell and/or tissue imaging, biological sample preparation methods and associated data analysis and exploitation.

Complementary knowledge and skills 

  • Experience in micro-fluidics, micro-fabrication and associated experimental environments.
  • Previous experience in cryogenics will be greatly appreciated.

Skills required

  • Time management.
  • Good communication skills with other CLEXM partners and users.

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Application Deadline: 18th August 2023 (Closed)