Live Fluorescence Imaging of Medaka Hatchlings

Live fluorescence imaging was performed using a stereomicroscope (Nikon SMZ18) equipped with the NIS-Elements BR 3.0 software or confocal microscopes (Zeiss Meta 500; Olympus FluoView FV3000; Zeiss LSM900). Medaka hatchlings (8 to 23 dpf) were anaesthetized with 0.005% ethyl 3-aminobenzoate methane sulfonate (Tricaine; Sigma MS-222) and mounted in 1.5% low-melting-point agarose on a glass bottom petri dish. Confocal pictures were taken using 405, 488, 543 or 633 nm laser lines for CFP, GFP, mCherry and Cy5 fluorescent signals, respectively. Time-lapse imaging was performed with Olympus FV3000 or Zeiss LSM900 microscopes by imaging the region of interest for 15-20 hours with 5-10 mins intervals. Imaging data were processed using Olympus FV31S-SW 2.1.1.98, Bitplane Imaris 9.0, ImageJ and Adobe Photoshop CC 2018 software.

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Imangali N., Sokolova V., Kostka K., Epple M, & Winkler C. (2023). Functionalized calcium phosphate nanoparticles to direct osteoprotegerin to bone lesion sites in a medaka (Oryzias latipes) osteoporosis model. Frontiers in Endocrinology, 14, 1101758.

Publication 2023

SMZ18 LSM900 MS-222 FV3000

Agarose Br elements Confocal microscopes Dish Ethyl methane sulfonate Medaka Microscopes Ms 222 Tricaine

Corresponding Organization : National University of Singapore

Other organizations : University of Duisburg-Essen

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Variable analysis

independent variables

Anaesthetized medaka hatchlings (8 to 23 dpf)

dependent variables

Fluorescence signals (CFP, GFP, mCherry, Cy5)
Time-lapse imaging of the region of interest for 15-20 hours with 5-10 mins intervals

control variables

Mounting medaka hatchlings in 1.5% low-melting-point agarose on a glass bottom petri dish
Anaesthetizing medaka hatchlings with 0.005% ethyl 3-aminobenzoate methane sulfonate (Tricaine; Sigma MS-222)

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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