Biology Reference
In-Depth Information
fully hydrated before staining and air-dried afterwards before
mounting into nonaqueous mounts. Acetate buffer pH 4.4
can be used instead of water to prepare dye solution for more
consistent results.
3. To reduce aldehydes on sections, dissolve 5 mg of NaBH
4
in
10 ml of borate buffer (pH 7.6) and treat section for 1 h in lab
temperature [
4
].
4. There are several pitfalls of the Sudan red staining procedure.
First, there might be problem with unspecifi c precipitation of
Sudan red pigment on sections. The primary reason might be
in water contact with the dyeing solution, which might pro-
duce crystals of Sudan. The dyeing solutions remain stable for
considerable period of time (months), but is sensitive to water
absorption and deteriorates if let open for a long time. We
have also experienced staining problems dues to long-term
storage (several years) of PEG 400 used for preparation of the
solution. Be also careful with microscope setup to localize well
cell wall response as plasma membrane staining may in some
cases cause seeming coloration of cell walls due to refraction.
The possibility of nonspecifi c staining of strongly acid struc-
tures (chromosomes) was indicated by Lillie [
47
].
Acknowledgment
This work has been supported by the MSM0021620858 project
and COST- LD11017.
References
1. O'Brien TP, Feder N, McCully ME (1964)
Polychromatic staining of plant cell walls by
toluidine blue O. Protoplasma 59:368-373
2. Sylvén BENG (1954) Metachromatic dye-
substrate interactions. Q J Microsc Sci
93-95:327-358
3. Bergeron JA, Singer M (1958) Metachromasy:
an experimental and theoretical reevaluation.
J Biophys Biochem Cytol 4:433-457
4. Pearse AG (1985) Histochemistry (theoretical
and applied). Churchill Livingstone, Edinburgh
5. Rost FWD (1995) Fluorescence microscopy.
Cambridge University Press, Cambridge
6. Kasten FH, Burton VIVI, Glover PEGG (1959)
Fluorescent Schiff-type reagents for cytochemi-
cal detection of polyaldehyde moieties in sec-
tions and smears. Nature 184:1797-1798
7. Truernit E, Bauby H, Dubreucq B et al (2008)
High-resolution whole-mount imaging of
three-dimensional tissue organization and
gene expression enables the study of phloem
development and structure in Arabidopsis.
Plant Cell 20:1494-1503
8. Herth W, Schnepf E (1980) The fl uoro-
chrome, calcofl uor white, binds oriented to
structural polysaccharide fi brils. Protoplasma
105:129-133
9. Wood PJ, Fulcher RG, Stone BA (1983)
Studies on the specifi city of interaction of
cereal cell wall components with Congo
Red and Calcofl uor. Specifi c detection and
histochemistry of (1-3), (1-4), -
β
-D-glucan.
J Cereal Sci 1:95-110
10. Benes K (1968) On the stainability of plant cell
walls with alcian blue. Biol Plant 10:334-346
11. Luft JH (1971) Ruthenium red and violet. I.
Chemistry, purifi cation, methods of use for
electron microscopy and mechanism of action.
Anat Rec 171:347-368
12. Muhlethaler K (1950) Electron microscopy of
developing plant cell walls. Biochim Biophys
Acta 5:1-9
Search WWH ::
Custom Search