Wiesner Staining for Visualizing Lignin: A Technique to Stain Lignin Deposits Present in Stems of Arabidopsis Thaliana

JoVE Encyclopedia of Experiments

Biologische Technieken

0 weergaven • 2:30 min • July 8th, 2025

$$\rightleftharpoonup{xx}$$ $$\longleftharp{xx}$$, $$\longrightharp{xx}$$,

In plants, lignin - a complex biopolymer - is typically deposited in the cell walls of the tissues to provide support.

Lignification is predominantly found in the secondary cell wall thickenings of water-conducting xylem cells, such as vessels and fibers, and the extra-xylary cells, such as interfascicular fibers.

Structurally, lignin is composed of crosslinked monolignol precursors. These aromatic alcohols polymerize, forming a highly branched structure. This feature enhances the plant’s strength and rigidity.

To visualize lignin deposition using Wiesner staining, take agarose-embedded dicot stem cross-sections derived from the plant’s middle or lower portion. Add them to a microcentrifuge tube containing a suitable buffer.

Add the prepared stain solution, which contains the acidified phloroglucinol and ethanol, to the stem sections. The alcohols present in the lignin react with the phloroglucinol and ethanol to yield a pink pigment. This results in the development of pink coloration corresponding to the lignin deposition in the cell walls of specialized cells.

Now, transfer the stained section onto a microscopic slide. Gently place a coverslip over the section, avoiding bubbles, to help better visualize it. Visualize the sections under a light microscope.

The pink coloration in the specialized xylem cells and the interfascicular fibers in the stem cross-sections confirm the presence of lignin in these tissues.

For Phloroglucinol staining, transfer the stem sections to a microcentrifuge tube. Add 1 milliliter of the phloroglucinol solution to the tube, and cap it immediately because the hydrochloric acid in the phloroglucinol stain is highly corrosive. Gently move the tube to ensure that all the sections are stained. Gently pipette the sections into a cut pipette tip, and onto a microscope slide. Cover the sections with a coverslip. Observe the sections under bright-field lighting. The solution dries up in 5 to 10 minutes causing a deterioration of the specimens, therefore, the imaging has to be completed within that period of time.

10:40

Visualiseren van Lignification Dynamics in planten met Klik op chemie: Dual Labeling is BLISS!

Gerelateerde video's

0 Bekeken

07:15

Meten van interacties tussen fluorescentie sondes en lignine in plantendelen door sFLIM op basis van inheemse Autofluorescentie

Gerelateerde video's

0 Bekeken

10:14

Fluorescerende immunolocalisatie van Arabinogalactan-eiwitten en pectines in de celwand van plantenweefsels

Gerelateerde video's

0 Bekeken

07:35

Label-free In situ Imaging van Lignification in Plant celwanden

Gerelateerde video's

0 Bekeken

02:44

Maule Staining for Lignin: A Technique to Characterize Lignin Distribution in Arabidopsis Thaliana Stem Sections

Gerelateerde video's

0 Bekeken

02:19

Congo Red Staining: A Technique to Visualize Cellulose Deposits in Plant Stem Sections

Gerelateerde video's

0 Bekeken

02:37

Calcofluor White Staining: A Simple Method for UV-Induced Fluorescence Visualization of Cellulose Deposits in Cell Walls of Dicot Plant Stem Sections

Gerelateerde video's

0 Bekeken

10:39

Histochemische kleuring van Arabidopsis thaliana Secundaire celwand Elements

Gerelateerde video's

0 Bekeken

13:00

Studeren Wnt Signalering Tijdens patroonvorming van Conducting Airways

Gerelateerde video's

0 Bekeken

07:51

Combinatie van Raman Imaging en Multivariate Analysis om Lignine, Cellulose en Hemicellulose in de Plant Cell Wall te visualiseren

Gerelateerde video's

0 Bekeken

Laatst bijgewerkt: 22 augustus 2026