Table 3.

Steps for the dehydration and the embedding with London Resin White of plant materials prepared for the analysis of plant cell walls by immunomicroscopy.

	Soft			Hard
Step	Solution	Incubation time	Temp.	Solution	Incubation time	Temp.
WASHES AFTER FIXATION
a	Buffer solution used to fix the samples	3 × 5 min	4°C	Buffer solution used to fix the samples	3 × 5 min	RT
b	dH2O	5 min	4°C	dH2O	5 min	RT
DEHYDRATATION
c	10% (v/v) ethanol/dH2O	10 min	4°C	10% (v/v) ethanol/dH2O	10 min	RT
d	20% (v/v) ethanol/dH2O	10 min	0°C	20% (v/v) ethanol/dH2O	10 min	RT
e	30% (v/v) ethanol/dH2O	10 min	−20°C	30% (v/v) ethanol/dH2O	10 min	RT
f	50% (v/v) ethanol/dH2O	10 min	−20°C	50% (v/v) ethanol/dH2O	10 min	RT
g	70% (v/v) ethanol/dH2O	30 min	−20°C	70% (v/v) ethanol/dH2O	30 min	RT
h	90% (v/v) ethanol/dH2O	30 min	−20°C	90% (v/v) ethanol/dH2O	30 min	RT
i	100% ethanol	30 min	−20°C	100% ethanol	30 min	RT
j	100% ethanol	30 min	−20°C	100% ethanol	O/N	RT
k	100% ethanol	30 min	−20°C
INFILTRATION
l	10% (v/v) LR White/ethanol	1 h	−20°C	10% (v/v) LR White/ethanol	1 h	RT
m	30% (v/v) LR White/ethanol	1 h	−20°C	30% (v/v) LR White/ethanol	1 h	RT
n	50% (v/v) LR White/ethanol	1 h	−20°C	50% (v/v) LR White/ethanol	1 h	RT
o	70% (v/v) LR White/ethanol	1 h	−20°C	70% (v/v) LR White/ethanol	1 h	RT
p	90% (v/v) LR White/ethanol	1 h	−20°C	90% (v/v) LR White/ethanol	1 h	RT
q	100% LR White	8 h	−20°C	100% LR White	O/N	4°C
r	100% LR White	8 h	−20°C	100% LR White	8 h	4°C
s	100% LR White	8 h	−20°C

On the left, the protocol termed “soft” is used for embedding soft plant materials such as the apical region of A. thaliana stems. On the right, the protocol termed “hard” is used for embedding hard plant materials such as strongly lignified A. thaliana stems or silicate-rich rice stem internodes. O/N, overnight; RT, room temperature, LR, London resin, dH₂O, distilled water.