Practical 3

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The experiment aims to observe microscopic structures of cinnamon bark, fennel fruit and clove flower bud using different staining techniques.

Lignified cells like pericyclic fibers, stone cells and cork cells; starch and calcium oxalate crystals were observed in cinnamon bark.

Phloroglucinol with HCl stained lignified structures pink while alcoholic picric acid solution stained aleurone grains yellow and Sudan red-III stained oil globules red in fennel fruit.

Practical 3: Transverse section of cinnamon bark, fennel fruit, and clove flower bud as well as the characteristic of cinnamon

powder. Aim: a) To perform the transverse sections of cinnamon bark and observe its microscopic characteristics (lignified cells, pericyclic fibers, stone cells, starch, calcium oxalate crystals and cork cells). In addition, the characteristics of cinnamon powder are also observed. b) To perform the transverse section of fennel fruit and observe its microscopic characteristics (lignified reticulate parenchyma of mesocarp and vascular bundles, aleurone grains, and oil globules in the cells of endosperm and cuticle). c) To perform the transverse sections of clove flower bud and observe its microscopic characteristics (vascular bundles and fibers, eugenol of volatile oil, cuticle, oil glands, calcium oxalate crystals and tannins in cortex region).

Apparatus and materials: Watch glass, knife, microscope, microscope glass, brush, cinnamon bark, cinnamon powder, potatoes, fennel fruit, clove flower bud, distilled water, phloroglucinol solution, conc. HCl, iodine solution, glacial acetic acid, alcoholic picric acid solution, Sudan red-III solution, iodine solution, dilute potassium hydroxide solution, dilute hydrochloric acid, ferric chloride solution (5%) and glycerine.

Procedures: *Glycerine was added to each microscopic glass to provide a clearer observation under a microscope. Experiment 3a: Transverse section of cinnamon bark 1. Cinnamon bark was cut into small thin sections. 2. A thin section of cinnamon bark was transferred to the microscopic glass with a brush. 3. One drop of phloroglucinol, one drop of concentrated HCl and a drop of glycerine were added to the thin layer. 4. It was placed under a microscope and observed. All the observation and results were recorded. 5. Step 2 & 3 were repeated using iodine solution as first and glacial acetic acid as the next. 6. Cinnamon powder was taken and mixed with one drop of phloroglucinol and one drop of concentrated HCl. 7. It was placed on the microscopic glass with glycerine on it and observed under the microscope. 8. The observation and result were recorded.

Experiment 3b: Transverse section of fennel fruit 1. A piece of fennel fruit was placed in the middle of the potato block as a support to ease the cutting procedure. 2. The thin section of fennel fruit was transferred to the microscopic glass with a brush. One drop of phloroglucinol, one drop of concentrated HCl and a drop of glycerine were added to the thin layer. 3. It was placed under a microscope and observed. All the observation and results were recorded. 4. Step 2 & 3 were repeated using alcoholic picric acid solution as first and Sudan red-III solution as the next. Experiment 3c: Transverse section of clove flower bud 1. Clove flower bud was cut into small thin sections. 2. A thin section of clove flower bud was transferred to the microscopic glass with a brush. One drop of phloroglucinol, one drop of concentrated HCl and a drop of glycerine were added to the thin layer. 3. It was placed under a microscope and observed. All the observation and results were recorded. 4. Step 2 & 3 were repeated for another 4 times using potassium hydroxide solution as first, Sudan red-III solution as second, dilute HCl as third and ferric chloride solution (5%) as the fourth.

Observations: (3a) Transverse section of cinnamon bark No Reagents 1 1 drop of phloroglucinol & 1 drop of conc. HCl Observation under microscope Characteristics Lignified cells: pericyclic fibers, stone cells, cork cells. Pink colour

1 drop of iodine solution

Starch. Blue colour

Glacial acetic acid

Calcium oxalate crystals. Insoluble

Cinnamon Powder 4 1 drop of phloroglucinol & 1 drop of conc. HCl Cork cells, phloem fibers and oil globule

(3b) Transverse section of fennel fruit No Reagents 1 1 drop of phloroglucinol & 1 drop of conc. HCl Observation under microscope Characteristics Lignified reticulate parenchyma of mesocarp and vascular bundles. Pink colour

1 drop of alcoholic picric acid solution

Aleurone grains. Yellow colour

Sudan red-III solution

Oil globules in the cells of endosperm and cuticle. Red colour

(3c) Transverse section of clove flower bud No Reagents 1 1 drop of phloroglucinol & 1 drop of conc. HCl Observation under microscope Characteristics Vascular bundles and fibers. Pink colour

1 drop of potassium hydroxide solution

Eugenol of volatile oil. Needle shaped potassium eugenate crystals are seen.

Sudan red-III solution

Cuticle, oil glands. Red colour

1 drop of dilute HCl

Calcium oxalate crystals. Soluble crystals

1 drop of ferric chloride solution (5%)

Tannins in cortex region. Dark colour

Conclusion: It can be concluded that, for the transverse section of cinnamon bark and cinnamon powder, the microscopic characteristics were observed under the microscope as followed:Test 1: Lignified cells such as pericyclic fibers, stone cells and cork cells were observed as pink colour. Test 2: starch (blue colour) Test 3: insoluble calcium oxalate crystals. Test 4: cork cells, phloem fibers and oil globule. For transverse section of fennel fruit, the following microscopic characteristics were observed:Test 1: lignified xylem reticulate parenchyma of mesocarp and vascular bundles were observed in pink colour. Test 2: Aleurone grains which is blue in colour. Test 3: Oil globules in the cells of endosperm and cuticle which were observed as red colour For transverse section of clove flower bud, the following characteristics were observed under the microscope:Test 1: pink colour vascular bundles and fibers Test 2: Eugenol of volatile oil and needle shaped potassium eugenate crystals are seen Test 3: red colour cuticle and oil glands. Test 4: soluble crystals of calcium oxalate crystals Test 5: Tannins in cortex region which is in dark colour

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