Table 1. Immunofluorescence staining protocol.

	Control group	Mixing group at each frequency
Fixation of eggs	Room temperature	Room temperature
30 min	30 min
Primary antibody	Rest in the droplet	AC electric field mixing
5 min	5 min
Washing	Room temperature	Room temperature
1 min	1 min
Secondary antibody	Rest in the droplet	AC electric field mixing
5 min	5 min
Washing	Room temperature	Room temperature
1 min	1 min
Mounting of eggs	Room temperature	Room temperature
VECTASHIELD®	VECTASHIELD®
30 min	30 min
First observation	Room temperature	Room temperature
<1 min per egg	<1 min per egg
Store of specimens	Dark room at 4 °C	Dark room at 4 °C
48 h	48 h
Second observation	Room temperature	Room temperature
for fading	<1 min per egg	<1 min per egg

AC, alternating-current; VECTASHIELD®, VECTASHIELD® Mounting Medium with DAPI.