Table 1.

Procedures and time for immunohistochemical staining

	Routine IHC procedure	Saving AC electric IHC procedure	Rapid AC electric IHC procedure
Acetone fixation	4°C	Room temperature	Room temperature
	10 min	2 min	2 min
Washing with PBS	5 min	30 sec	30 sec
	3 times	once	once
Primary antibody	w/o AC electric field	AC electric field	AC electric field
	60 min	60–180 min	2 min
Concentration of primary antibody	5.0 µg/mL	0.5–2.0 µg/mL	5.0 µg/mL
	(1:200)	(1:500–2,000)	(1:200)
Washing with PBS	5 min	30 sec	30 sec
	3 times	once	once
EnVisionTM+ System/HRP Mouse	w/o AC electric field	AC electric field	AC electric field
	30 min	2 min	2 min
Washing with PBS	5 min	30 sec	30 sec
	3 times	once	once
3,3'diaminobenzidine	3 min	3 min	3 min
Approximate time required	148 min	69–189 min	11 min