Accelerated image acquisition applied on acidian embryos exposed to genotoxics and endocrine disruptors in the sea

 

Rémi Dumollard is a researcher at the Developmental Biology Laboratory of Villefranche-sur-mer, France.
Working on his project Marine-EmbryoTox, Rémi tries to achieve a molecular test based on Transgenic Ascidian Embryos to detect pollution caused by endocrine disruptors existing in the sea.

Using Multi-Dimensional Technique to observe the cellular dynamics

Rémi uses the multi-dimensional imaging technique on fluorescently‐tagged proteins to follow the dynamics of cellular structures. The transgenesis of XFP-based probes (CFP, GFP, Venus, Cherry, mRFP, dTomato) processed inside ascidian embryos allows to generate hundreds of fluorescent embryos in order to detect genotoxics and endocrine disruptors. These ascidian embryos are put down a 96-well plate.

The entire embryonic development takes 16 hours. During all this time, samples are imaged in high content screening conditions : 3 dimensions and 3 channels for each embryo.

More images in same time to finely follow the sample development

For his first tests, Rémi had to use an epifluorescence imaging station controlled by a conventional automation and image acquisition software. This system was able to screen up to 25 embryos for the 16 hours timelapse.

The usefulness of a dataset for further analysis is very dependant of the number of images acquired during these 16 hours. With its new station controlled by Inscoper hardware-based solution, Rémi now ensures the screening of up to 96 embryos in same conditions, a far better performance for his research.
Complete technical characteristics of this imaging station are shown in the table .

MicroscopeLeica DMi8
TypeInverted
IlluminationEL6000 HXP120 100W
CameraHamamatsu Orca Spark
SoftwareInscoper Imaging Suite
Objectives

5x/0,12 N PLAN

20x/0.40 HCX PL Fluotar

10x/0.32 HC PL Fluotar

Reflector

RHOD LP excitation 540/45 nm, dichroic 580 nm, emission LP590 nm

YFP excitation 500/20 nm, dichroic 515 nm, emission 535/30 nm

CFP excitation 436/20 nm, dichroic 455 nm, emission 480/40 nm

Filter wheel

CFP, excitation : 436/20 nm,

YFP, excitation 500/20 nm,

RHOD LP, excitation 540/45 nm

Applications

Multi-dimensional acquisition :

  • Timelapse
  • Multi-positions
  • Multi-Z
  • Multi-channels
  • 96-well plate