3.6  Visualization of cell walls with calcofluor


Tomomichi Fujita and Takashi Murata


Calcofluor binds cellulose in cell walls and is used to visualize cell walls. It emits blue-white fluorescence under UV excitation.


Solution preparation

1. 1 mg/ml fluorescent brightener 28 (calcofluor)


Stock solution    

fluorescent brightener 28 (sigma, F-3397)                1 mg   

H2O                                     1 ml      

  Dissolve, then store at -20˚C covered with aluminum foil to prevent light.


2. fluorescent brightener 28 solution

  Dilute the stock solution one hundred-fold (10 ug/ml)

   1 mg/ml fluorescent brightener 28           10 ul   

   H2O                                  1 ml      

   Store at –20˚C.



Place tissues or cells onto a glass slide, and drop the calcofluor working solution onto them. Wash the preparation with PBS. Replace PBS with distilled water as quickly as possible and observe under UV excitation immediately.

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(Figure) The first asymmetric cell division of a P. patens protoplast. Cells in bright field (upper) and with calcofluor under UV field (lower).