Recently I am working on chloroplast in Mesophyll cell of Rice Leaf, I have done semi-thin section of Rice Leaf and tried Toluidine Blue O but result was not so clear. I would like to know which technique is suitable for chloroplast staining.
Dear Hayat,
Here is a procedure as extracted from the article cited at the end...
Leaf tissue from each zone was fixed whole in 5% (v /v) glutaraldehyde in Sorensen’s phosphate buffer, pH 7.2. Leaves were sectioned into 1-mm wide strips while submerged in glutaraldehyde. Sections were stored at 4 °C. Serial dehydrations in 30%, 50%, 75%, 95%, and 100% ethanol were performed, followed by four 15-min washes with propylene oxide. Araldite 502 resin was used to embed samples in flat molds. Thick sections (ca. 400 nm), obtained using a Porter-Blum MT-2 ultramicrotome, were stained with 0.5% toluidine blue in 1% borax. Thin sections, obtained using an Ultratome Nova, were stained with saturated aqueous lead citrate and uranyl acetate. Transmission electron micrographs were obtained using a Joel 1200 Ex II electron microscope. Fifty individual chloroplasts were scored from each zone to determine distribution of chloroplast types at different stages of senescence.
(Plant Biol (Stuttg). 2010 Jan; 12(1): 1–12. doi: 10.1111/j.1438-8677.2009.00206.x)
Good luck!
Article:
Electron-cytochemical osmium-ferricyanide staining of cereal chloroplasts
February 1987Protoplasma 140(1):22-25
DOI10.1007/BF01273252
V. V. MosejevO. I. RomanovskayaV. V. Iljin
Would be glad if anyone could help me with recommended dye or stain for viewing the structure of chloroplast under microscope in peels. Such as pear peel during storage.
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