Common facilities

Team Leader :
Myriam MOHAMED



 

Missions 


The EpHISTain facility is dedicated to the technical support of the teams of the “Epigenetics and Cell Fate” Unit in the fields of histology and cytology in order to support the research work of the teams in the elucidation of the role, the mechanisms of implementation and maintenance of epigenetic modifications that determine plasticity and cell fate.

This service sets up various histological techniques, histochemistry, immunohistochemistry, immunofluorescence, in situ hybridization (ISH), fluorescence in situ hybridization (FISH) and cytology, for biological tissue sections production and staining (animal or organoid), as well as cells labelling.

In the near future, it is also moving towards the implementation of conventional and molecular cytogenetic techniques (karyotype and FISH) to routinely control the quality of cells before and after editing the (epi) genome.

 

Prestations


  1. Technical support or sample handling (fresh, fixed, frozen) for the biological tissue sections production. The chosen technique depends on the technical and scientific teams’ questions, as well as the treatments necessary to answer them:
  • Tissue dissection, preparation and fixation (including embryonic)
  • Tissue embedding (OCT and agarose)
  • Tissue sectioning (vibratome or cryostat), on slides or in floating sections 

  1. Biological tissue sections treatment. The treatment choice depends on the type of tissue, the technical issues and the biological question to be answered by the experiment:
  • Explore tissues morphology or the structures organization of interest on sections in order to highlight possible modifications or structural defects
    • Histochemical staining on biological tissue sections (e.g. cresyl violet)
    • In Situ Hybridization (ISH) on cryostat sections, floating or on slides, for the labelling of specific structures (e.g.: cortical layers markers)

    

  • Characterize the expression profile of genes of interest (expression or not and cellular localization) by targeting RNAs or proteins
    • In Situ Hybridization (ISH) on cryostat sections, floating or on slides
    • Immunohistochemistry and Immunofluorescence
    • FISH (DNA & RNA)
  1. Ensure a quality check of stem cells before and after (epi)genome editing (service under development)
  • Genomic and transcriptional landmarks of “normal” cellular behavior (e.g. the X chromosome, satellite sequences…)
    • DNA FISH (Fluorescent in situ hybridization)
    • RNA FISH
  • Karyotyping of these stem cells
    • Chromosome spreading
    • Chromosome painting
    • Spectral karyotyping (SKY) FISH
  1. Imaging of quality check results (on the EPI2 platform)
  1. Support
  • Optimize and standardize the labeling protocols on tissues and cells for homogeneity and reproducibility which allow the teams’ data to be crossed.
  • Adapt and develop staining protocols on tissues or cells according to the specific questions of the teams, by providing technical expertise
  • Train team members in cutting and labelling techniques as needed, sharing protocols and know-how.

 

Members & Steering Committee


For any information request or to submit a project, you can contact

Myriam Mohamed – CNRS Technician

Team : Common service

myriame.mohamed@u-paris.fr

 

Any new project is submitted for validation to the steering committee which scientifically supports the platform

 

Véronique Dubreuil – MCF U-PARIS

Team : Development and environment interface

 

Claire Francastel – Team leader / DR2 INSERM 

Team : DNA methylation and non-coding RNA in health and disease

 

Céline Morey – CR1 INSERM

Team : Non-coding RNAs, differentiation and development