2.6 Cell death and proliferation staining of A. erythraea(Expt3.)
To further explore the effects of DA on copepods at the cellular level,
we first cultured A. erythraea on the two diets for 48h and then
fixed them with 4% paraformaldehyde in phosphate-buffered saline (PBS).
For the measurement of dead cells, the terminal deoxynucleotidyl
transferase biotin-dUTP nick end labelling (TUNEL) assay was performed
with an in situ cell death detection (TMR Red) kit according to the
manufacturer’s instructions. Labelling and detection of proliferating
cells in A. erythraea were performed using the Click-iT™ EdU Cell
Proliferation Kit (Invitrogen), with the following modifications.A. erythraea were incubated in a 10 μM solution of
5-Ethynyl-2-deoxyuridine (EdU) for 3 h immediately before fixation in a
solution of 4% paraformaldehyde in PBS. The copepods were viewed and
photographed with a Leica M205 FCA microscope equipped with a Leica DFC
7000T camera.
Results and Discussion