Digital sight qi2 monochrome camera

Cells in the mid-log phase of growth (1−5 × 10⁶ cells/ml) were grown in HL5 overnight to confluency in Petri dishes. The following day, cells were washed twice with KK2 buffer and then submerged in fresh KK2 buffer. Starving cells were imaged using a Nikon Ts2R-FL inverted microscope equipped with a Nikon Digital Sight Qi2 monochrome camera. Cells were also harvested after 4 h of starvation, lysed with NP40 lysis buffer, and analyzed by western blotting to assess the effect of cln5-deficiency on the intracellular and extracellular amounts of CadA, CtnA, and CtsD, as well as the intracellular and extracellular amounts of Cln5 in atg1^– and atg9^– cells. In separate experiments, 5 × 10⁶ cells were deposited into 6 well dishes and allowed to adhere for 30 min. Cells were then washed twice with FM-aa and incubated in fresh FM-aa for 24 h. Mounds were imaged after 24 h using a Nikon Ts2R-FL inverted microscope equipped with a Nikon Digital Sight Qi2 monochrome camera.