3.10 Visualization of cell wall with calcofluor
Tomomichi
Fujita
Calcofluor
binds cellulose in cell wall and is used to visualize cell wall. It emits
blue-white fluorescence under UV excitation.
1.
1 mg/ml fluorescent brightener 28 (calcofluor)
fluorescent brightener 28 (sigma、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 a stock solution
into one hundredth (10 ug/ml)
1 mg/ml fluorescent
brightener 28 10 ul
H2O 1 ml
Store at –20˚C.
Protocol
Drop
the calcofluor working solution onto tissues or cells, and observe under UV
excitation. Excess amount of calcofluor, which causes high background should be
washed with PBS buffer. Wash cells as quick as possible, otherwise cells are
subject to plasmolysis.
(Figure) The first
asymmetric cell division of P. patens protoplast. Cells in
bright field (upper) and with calcofluore under UV field (lower).