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Lignin Staining
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Materials Required

 

1. Potato
2. Double edged razor blade
3. Dropper
4.  Brush
5. Glass slide and coverslip
6. Needle
7. Phloroglucinol stain
8. Petridish
9. Watch glass
10. Stem of a plant.

 

  Preparation of Phloroglucinol-HCl Stain

 

There are various procedures to make up the staining solution but commonly it is prepared as a saturated solution of phloroglucinol in 20% hydrochloric acid. The hydrochloric acid used is about 2 N. Please be sure to handle the solution with care. Wear gloves. Prepare this solution in the fume hood. First dissolve phloroglucinol (about 2.0 g) in 80 ml of 20% ethanol solution and then add 20 ml of concentrated HCl (12 N) to it.

  

Procedure

 

 1. Take a potato and cut it into a block.  Make a slit in the middle of the potato block using a razor blade.
2. Place the stem of the plant in the middle of the slit.
3. Cut thin sections of the potato block along with the stem into a petridish containing water.
4. Transfer the only the cut sections of the stem using a brush into a petridish containing water.
5. Add two or three drops of phloroglucinol stain to a watchglass using a dropper.
6. Transfer the potato sections using a brush into the watch glass containing the stain.
7. Take a clean glass slide and add two drops of water onto the slide. Transfer the sections onto the slide using a wet brush.
8. Place a cover slip onto the slide with the help of a needle or forceps.
9. Remove the solution from the edges of the cover slip using a blotting paper and observe under the microscope.

 

Difference Encountered in a Real Laboratory


In an actual laboratory setting, there are certain important steps that are not necessarily applicable   in a virtual lab:


1.    Before starting the experiment sterile the laminar air flow chamber using spirit.
2.    Ensure that while placing the cover slip over the section.
3.    Avoid the presence of air bubbles in between the cover slip and section.
4.    Always disinfect your work area when you are finished.


 

 

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