Accelerated image acquisition of acidian embryos exposed to genotoxic substances and endocrine disruptors in the sea

Rémi Dumollard is a researcher at the Villefranche-sur-Mer Developmental Biology Laboratory (France).

Working on his Marine-EmbryoTox project, Rémi tries to conduct a molecular test on transgenic ascidian embryos to detect pollution caused by endocrine disruptors in the sea.

 

 

Using multidimensional imaging to observe cell dynamics

Rémi uses multidimensional imaging on fluorescently tagged proteins to observe the dynamics of cellular structures. Transgenesis of XFP-based probes (CFP, GFP, Venus, Cherry, mRFP, dTomato) in ascidian embryos can be used to generate hundreds of fluorescent embryos in order to detect genotoxic substances and endocrine disruptors. These ascidian embryos are put down into 96-well plate.

Complete embryonic development takes 16 hours. During this time, samples are imaged in high-content screening conditions (three-dimensional, 3 channels for each embryo).

 

 

More images in the same timeframe to closely follow sample development

For his first tests, Rémi had to use an epifluorescence imaging station controlled by conventional automation and image acquisition software. This system was able to screen up to 25 embryos during the 16 hour time-lapse.

The usefulness of a dataset for further analysis is highly dependent on the number of images acquired during these 16 hours. With the new station, controlled by the Inscoper hardware-based solution, Rémi can now screen up to 96 embryos in the same conditions, which is far better for his research.

Full technical characteristics of this imaging station are shown in the table below.

Microscope Leica DMi8
Type Inverted
Illumination EL6000 HXP120 100W
Camera Hamamatsu Orca Spark
Software Inscoper
Objectives 5x/0,12 N PLAN
20x/0.40 HCX PL Fluotar L
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