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Calcofluor White Staining: A Simple Method for UV-Induced Fluorescence Visualization of Cellulose Deposits in Cell Walls of Dicot Plant Stem Sections
Overview
In this video, we describe a staining procedure to visualize cellulose deposition within cell walls of Arabidopsis thaliana, a dicot plant, using calcofluor white, which specifically binds to β-linked glucans, exhibiting fluorescence under UV light.
Protocol
1. Stem Embedding
- Make a 7% agarose solution in water (7 g of electrophoresis-grade agarose in 100 ml distilled water). Dissolve the agarose by autoclaving for 20 min or by microwaving for 20 min at lowest intensity (e.g., 10% intensity of a 1,250 watt microwave).
- Prepare a homemade mold for embedding the stems using plastic vials.
- Using a razor blade, cut off the conical bottom of a 2 ml screw-cap microcentrifuge tube (Part A). With a syringe needle, puncture a.......
Representative Results
Figure 1. Homemade mold for embedding the stems. On the left side; top of the 2 ml screw-cap microcentrifuge tube (Part A) and the bottom from the 0.6 ml microcentrifuge tube (Part B). On the right side; the Parafilm holding the two parts; A and B to form the final mold.
References
This article has been published
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Source: Pradhan M. et al. Histochemical Staining of Arabidopsis thaliana Secondary Cell Wall Elements. J. Vis. Exp. (2014)
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