Plant cell wall staining and microscopy

RK Richa Kaushal
LP Li Peng
SS Sunil K. Singh
MZ Mengrui Zhang
XZ Xinlian Zhang
JV Juan I. Vílchez
ZW Zhen Wang
DH Danxia He
YY Yu Yang
SL Suhui Lv
ZX Zhongtian Xu
RM Rafael J. L. Morcillo
WW Wei Wang
WH Weichang Huang
PP Paul W. Paré
CS Chun-Peng Song
JZ Jian-Kang Zhu
RL Renyi Liu
WZ Wenxuan Zhong
PM Ping Ma
HZ Huiming Zhang
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To stain cellulose, Mitra and Loque [61] protocol for Calcofluor staining was followed with some modifications. Briefly, the roots were transferred to 2 ml tubes and stained with 0.02% calcoflour white (Sigma) for 5 min. The primary root was placed on the microscope glass slide and transverse section of middle part of the roots was prepared using sharp blade. Calcofluor White was visualized using an epifluorescence microscope (Zeiss Imager M2).

The staining of callose was done according to Muller et al. [62] with some modifications. Briefly, the roots were stained with 0.1% (w/v) aniline blue solution in 0.1 M sodium phosphate buffer (pH = 7.2) for 1.5 h. The cross-section of roots was done as described above and visualized under epifluorescence microscope (Zeiss Imager M2).

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