and 75 mL of the mixture were placed onto cover slip (24 50 mm), immediately a microscope pre-coated slidewas carefully connected avoiding to form air bubbles and placed on an ice surface for 5 min; then immersed in cold lysis solution (2.5 M NaCl, 100 mM EDTA, 10 mM Tris, 1% Triton X-100, and 10% DMSO)