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