In this video, we demonstrate the Congo red staining method to visualize cellulose deposits in Arabidopsis thaliana stem sections.
Protocol
1. Congo Red Staining Dissolve 0.5 g congo red in 100 ml of distilled water to prepare a 0.5% congo red solution. Transfer stem sections to a 2.0 ml microcentrifuge tube. Add 1 ml of the 0.5% congo red solution to the tube. Gently pipette the 0.5% congo red solution up and down without disturbing the sections. NOTE: Be careful not to pipette the sections into the pipette tip as this may damage the sections. Incubate at room temperature for 10 min and repeat the pipe…
Divulgazioni
The authors have nothing to disclose.
Materials
Congo Red
Sigma
C6277
Disodium 3,3'-[[1,1'-biphenyl]-4,4'-diylbis(azo)]bis(4-aminonaphthalene 1-sulphonate) [CAS Number 573-58-0]
Microcentrifuge Tubes (0.6 ml)
Axygen Scientific
MCT-060-C
Camera with CCD chip with no mechanical shutter
Hamamatsu
C4742-95
High speed color camera
QImaging
MicroPublisher 5.0 RTV
Camera software
Molecular Devices
MetaMorph version 7.7.0.0
Imagining anaylsis
Adobe
Photoshop CS4
Micro Cover Glasses, Square, No. 1
VWR
48366-067
22 x 22 mm (7/8 x 7/8")-Cover glasses are corrosion-resistant and uniformly thick and flat. No. 1 thickness is 0.13 to 0.17 mm.
Frosted Micro Slides, 1 mm
VWR
48312-003
75 x 25 mm – 1 mm
TX2 Filter cube
Leica
11513851/11513885
Filter used for Congo red analysis with a band-pass of 560/40.