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Experiment III

Microscopic Structure of Cells organelles
The Institute of Cell Biology, Medical College of Shandong University Teacher: li xia Dec.19/2002

1. Acquaint with some morphological characters of organelles by light microscopic. 2. To master the method and principle of vital staining in mitochondrion. 3. To learn the biological mapping method.

. Observation of organelles morphology by light microscope.

Figure 1: The structure of a eukaryotic cell

Schematic model of a portion of a Golgi complex from an epithelial cell of the male rat reproductive tract. . Electron micrograph of a portion of a tobacco root cap cell. B.Golgi complex----rabbit ganglion slice A.1.

Golgi complex Figure 3: You can see this map by 40× objective lens. .

0 µm in cross-sectional diameter and 1 to 4 µm in lengh. mitochondria are large enough to be seen in the light microscope. .it is 0.2. Approximately. Unlike most cytoplasmic organelles. Mitochondrion²² renal tubule slice Mitochondrion can produce chemical energy which is made available to fuel cellular activities.2 to 1.

. showing the internal matrix enclosed by folds of the inner membrane. (b) Schematic diagrams showing the generalized internal structure of a mitochondrion.a b Figure 4: The structure of a mitochondrion. (a) Scanning electron micrograph of a macerated mitochondrion.

Mitochondria are seen as elongated. particularly the membranous folds (cristae) of the inner membrane. (c) Localization of mitochondria in the sperm midpiece surrounding the proximal portion of the flagellum. .Figure 5: Mitochondria. (a) A living fibroblast viewed with a phase-contrast microscope. dark bodies. (b) Transmission electron micrograph of a thin section through a mitochondrion revealing the internal structure of the organelle.

.mitochondrions Figure 6: You can see this map by 40× objective lens.

. centrosome²² equine ascarid zygote slice In animal cells the centrosome is a complex structure that contains two barrel-shaped centrioles surrounded by amorphous.3.2 mm in diameter and typically about twice as long. electron-dense pericentfiolar material . Centrioles are cylindrical structures about 0.

which is undergoing elongation in this phase of the cell cycle . Each pair consists of a longer parental centriole and a smaller daughter centriole (arrow). (b) Electron micrograph of a cross section of a centriole showing the pinwheel arrangement of the nine peripheral fibrils. the surrounding pericentriolar material. (a) Schematic diagram of centrosome showing the paired centrioles. (c) Electron micrograph showing two pairs of centrioles. .Figure 7 Structure of the centrosome. each of which consists of one complete microtubule and two incomplete microtubules.

. which occurs in the two centrosomes located at opposite poles of a dividing cell.Figure 8: The role of tubulin in centrosome function. A dividing fibroblast that has been double stained with antibodies against -tubulin (red) and -tubulin (green). Orange staining is due to the coincidence of the two types of tubulin.

centrosphere centriole Figure 9: You can see these maps by 40× objective lens. .

4. The cytoskeleton is composed of three well-defined filamentous structures---microtubules. microfilaments. . and intermediate filaments--which together form an elaborate interactive network. cytoskeleton²² liver cancer cell or Hela slide The skeleton of a vertebrate is a familiar organ system consisting of hardened elements that support the soft tissues of the body and play a key role in mediating movements.

. The distribution of stress fibers in a cultured fibroblast as revealed by fluorecent anti-action antibodies(green).Figure 11: Stress fibers.

The celeste fibers aggregating by microfilament inweave net.Under high objective lens. . The cytoskeleton fibers is blue distributed in the cytoplasm. The nuclear is mazarine. the liver cells show polygon shape.

.Figure 12: You can see this map by 40× objective lens.

.5.It is important in the storage and utilization of genetic information. nucleus and nucleolus²² salamander epidermis slide The nucleus of a eukaryotic cell has a rather undistinguished morphology.

Figure 13 The cell nucleus. . (a) Electron micrograph of an interphase HeLa cell nucleus showing a pair of nucleoli and scattered chromatin. Heterochromatin is evident around the entire inner surface of the nuclear envelope. and clumps of chromatin can be seen scattered throughout the nucleoplasm. Two prominent nucleoli are visible. (b) Schematic drawing showing some of the major components of the nucleus.

They are nuclear membrane. . The chromatin was dyed in purple.Under high objective lens. the structure of nuclear are composed of four parts. chromatin. nucleolus is mazarine and nuclear matrix is not colouring. nucleolus and nuclear matrix.

nuclear neucleoli Figure 14 You can see this map by 40× objective lens. .

Make human oral cavity epidermal cell temporarily slide to show living mitochondrion.5²1 minute Then you can observe the slide after overlaying a cover glass. Manipulation 1. (1) method Take your oral cavity epidermal cells coating them on the clear microscopic slide droping one or two dyeliquor of Janus green B for 0. ..

Figure 15: The making process. .

(2) observation Under high objective lens----these brilliant green mitochondrions disperse in cytoplasm. Figure 16 . showing granulose or short bar.

choose useful things get into cell and keep harmful things outside. Permeability of cell membrane: Cell membrane can control exchanging substances with circumstance. .2.

but they can enter the dead cells and cause them colored because their membranes were destroyed . Many acid dyes can not pass through membrane of living cell. Principle: We use dyeing exclusive method to test this. .(1) Distinguish living and dead cells. So we can distinguish the living and dead cells. .

. put a then observe coverslip on the slide by your microscope. Method Dyeing cells with Typan Blue. Get a drop of yeast suspension drip on a clear slide dye-liquor then add a drop of after 1~2min..

dead cells Figure 17: Distinguish living and dead cells .

then calculate the cellular vitality. Result: The dead cells were dyed blue and the living cells were not colored. You may count one hundred cells you are observing.. cellular total -.the death number CellularVitality (%) = ×100% cellular total .

Because the plant cell has cellulose wall. Principle: ue to the different osmotic pressure between the internal and external. . Oppositely when we eliminate the hypertonia liquid and dilute it with water. they show a phenomenon of plasmolysis. water can pass through the membrane from low osmotic pressure to high place. We will see the phenomenon of deplasmolysis. the osmotic pressure is equal roughly between the internal and external. So when we put the cell in a condition of high osmotic pressure. the water will come out from the cell.(2) The plasmolysis and deplasmolysis: .

they gather in the center of cells. .To make an temporarily slide of onion epidermal cell (Experiment ). In the end. Put a drop of 20% sucrose solution (hypertonia liquid. then suck the water with a absorbent paper on the other side. you can see that the plasma depart from the cell wall gradually. b.. Method a. This is called the plasmolysis. separatory solution) on one side of the coverslip. Under low objective lens.

. then suck it on the other side for moving sucrose solution with a absorbent paper. rop some water on one side of the coverslip. drawing close to cell wall gradually. This is called the deplasmolysis. You can see the plasma extending outward again.c.

(a)Aquatic plants living in freshwater are surrounded by a hypotonic environment.creating turgor pressure.Figure 18: The effects of osmosis on a plant cell.such as seawater. . the cell loses water.Water therefore tends to flow into the cells. and the plasma membrane pulls away from the cell wall. (b) If the plant is placed in a hypertonic solution.

It must be authentic. . Biological drafting is a scientific record.Report: Drawing a map of oral cavity epidermal cell that you saw under the light microscope.

The outline of the shape must be showed by uniform and continuous line. . The structure and threedimensional effect must be showed by many spots. . The line can¶t be broke off or overlapped. .Method: . position and every regions ratio of your shape.Draw the map with 2H or 3H pencil. place . . Observe your samples seriously. You¶d better take care of its size. Draw some parallel lines on one or two sides for making names of every part.