Table 3. Optimization of immunolocalization of ZO-1, actin, hnRNP L, and histone H3 in endothelial cells on flat-mounted whole human corneas.

Target protein	Classification	Selected protocols
ZO-1	best	methanol at −20 °C
	second choice	methanol at RT; 0.5% formaldehyde at RT; Carnoy I at −20 °C
Actin	best	methanol at RT
	second choice	Carnoy I at −20 °C
hnRNP L	best	0.5% formaldehyde at RT
	second choice	methanol at RT
histone H3	best	0.5% formaldehyde + AR with 0.5% SDS in water 5 min at RT *
	second choice	methanol at RT

* SDS: Sodium dodecyl sulfate. In these cases fixation was prolonged during 1 h to avoid endothelial cell detachment induced by SDS with short time fixation. For all other cases, fixation during 30 min.