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  • A catalog of more than 300 available antibodies
  • Fine tuning of new antibodies
  • Chromogenic and fluorescence protocols

 In Situ Hybridization

  • On Slides
  • In toto

 Fields of study

  • Demonstration of transcripts of embryos or whole larvae.

 Technical interests

  • To locate the expression of a gene on a whole animal (larvae or embryo).
  • Demonstration of localization and level of expression of protein of interest indirectly by corresponding RNA (exotic species for which no antibody is available).
  • Automation with Intavis® device
The in toto In Situ Hybridization technique consists in performing the probes’ incubations directly on embryos fixed and permeabilized with proteinase K.

All manipulations are performed in thermostatically controlled cups with embryo-adapted sizes. Incubation times and medium changes are standardized on the Intavis® equipment.

The observation enables to precisely locate the expression of RNA and to evaluate its level of expression.

Results are transposable to the corresponding protein.

It is then possible to perform sections of marked area directly on the embryo by using a vibrating microtome.

Fields of study

This technique is mainly used on brain tissues. Series sections are performed on brains which are previously fixed and then frozen by using isopentane.
The purpose is to quantify glial cells which are previously stained by using markers such as Thyrosine Hydroxylase in specific area of the brain. (black substance).

Technical interests

  • This technique enables to quantify immunostained neurons in the complete area of interest (eg black substance thanks to series sections).
  • It should be performed manually and carefully. Sections are incubated in wells and then deposit on glass slides with a specific coating.
  • This reduces handlings and risks to lose samples through series sections.
This technique requires a good knowledge of brain’s anatomy of species of interest in order to perfectly locate the sections.
The analysis could be performed through stereology.

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