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Technical platforms

High throughput screening of strains


To select the strains of interest, the laboratory has developed two screening methods. The first one discriminates on the lipid composition, the second on the photosynthetic activity of the organism.

Published on 6 November 2019

​Screening of a mutant bank on the lipids content

 

​Step 1 :

Constitution of mutants banks
                             

Credit : G. Lesénéchal/CEA

Step 2 :

Cellular culture









Credit : G. Lesénéchal/CEA
​In microplates of 96 wells

​Step 3 :

​Screening of cells









Credit : CEA
Screening of the cells which accumulate TAG with flow cytometry (Use of a fluorescent lipophile marker: the Nile Red).

​Step 4 :

​Validation
by HPTLC



Credit : CEA
 Validation by HPTLC (High Performance Thin Layer Chromatography) Extraction of the lipids in microplates.

​Step 5 :

Evaluation of the lipid profils

Credit : CEA

By GC-FID or GC-M

 

Publication of the screening method :

Cagnon C, Mirabella B, Nguyen HM, Beyly-Adriano A, Bouvet S, Cuiné S, Beisson F, Peltier G, Li-Beisson Y. A genetic screen to isolate oil mutants in the green microalga Chlamydomonas reinhardtii . Biotechnol. Biofuels 6, 178

 

 

Screening of a mutant bank on the chlorophyll fluorescence

Credit : B. Gentil/CEA


A system allowing to follow the chlorophyll fluorescence in time, on plates of culture, was finalized with the cooperation of Bernard Genty and Illia Reiters (LEMP-CEA Cadarache).

This device based on the acquisition by a fluorescent camera of the signal of fluorescence and an excitement by a crown of diodes allowed to screen a bank of more than 3000 mutants of Chlamydomonas to isolate photosynthesis mutants (Ghysel and al .).