Caulerpa is a giant siphonous alga with a complex morphology organised within a single
multinucleate cell, making it an interesting but technically challenging system for internal
imaging. This thesis developed microscopy‑based workflows, from sample preparation to
imaging and data processing, to improve visualisation of internal morphology in thick Caulerpa
tissues, focusing mainly on Caulerpa prolifera and, on a smaller exploratory scale, Caulerpa
verticillata. Spectroscopy was used to characterise absorption spectra and identify excitation
wavelengths that penetrate tissues most effectively. Widefield microscopy provided rapid
overviews of tissue architecture and regeneration, while fluorescence and light‑sheet
microscopy allowed visualisation of nuclear and actin‑rich structures in cleared samples,
revealing internal organisation usually obscured by pigmentation. Two clearing methods,
CUBIC and benzyl alcohol benzyl benzoate (BABB), were tested in combination with nuclear
and actin stains (Propidium Iodide, Nuclear Green, Phalloidin 488, Phalloidin Texas Red).
Their performance was evaluated across multiple imaging platforms. Both clearing
approaches improved internal visualisation. However, Nuclear Green produced the most
useful insight, especially in combination with Phalloidin Texas Red in BABB. Overall, the study
provides a proof of concept for whole‑mount imaging in Caulerpa and a practical basis for
future work on organelle distribution and intracellular organisation in siphonous algae.
promotor/supervisor feedback
nothing yet
