Human Lymph Node

2um thick human lymph node, serial sections
Primary Abs: overnight @4C
Mouse @CD20-eFluor660 1:50
Rabbit @CD3 1:200
Secondary Ab: 1h RT, Donkey anti – rabbit IgG-PerCP (711-126-152),
Image acquisition:
Leica Upright SP8 confocal microscope.
PerCP excitation: 488nm laser intensity 3.4%, detection by HyD 144%
eFluor660 excitation: 638nm laser intensity 0.4%, detection by HyD 32%
DAPI excitation: 405nm laser intesity 0.2%, detection by HyD 50%