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11310386

2014.4.18

The Cell Cycle and Mitosis

The Cell Cycle and Mitosis

Introduction
The ordered sequence of events that extends from the time a cell is first formed from a dividing parent cell until its own division into two cells is called the cell cycle. Most of the cell cycle is spent in interphase. Interphase is a time when a cell performs its normal functions within the organism. During interphase, a cell roughly doubles everything in its cytoplasm. From the standpoint of cell reproduction, the most important event of interphase is chromosome duplication, when the DNA in the nucleus is precisely doubled. The period when this occur is called the S phase. The interphase periods before and after the S phase are called the G1 and G2 phases, respectively. During G2, each chromosome in the cell consists of two identical sister chromatids, and the cell during this period is preparing to divide. The part of the cell cycle when the cell is actually dividing is called the mitotic(M) phase. It includes two overlapping stages, mitosis and cytokinesis. There are four main stages of mitosis: prophase, metaphase, anaphase, and telophase. In mitosis, the nucleus and its contents, most importantly the duplicated chromosomes, divide and are evenly distributed, forming two daughter nuclei. During cytokinesis, the cytoplasm is divided in two. The combination of mitosis and cytokinesis produces two genetically identical daughter cells.

Methods and Materials

Methods: Compound microscopes(Nikon-Eclipse-E100-LED), water bath 60! Glacial acetic acidmethanol fixative(1:3), 1M HCl, 45% C2H6COOH, Feulgen stain; Prepared slides of onion root tip, lens tissue, garlic root tips, forceps, slides, coverslips. Methods: 1.Observe mitosis in plant(cells onion root tip) 1)Obtain a prepared slide of a longitudinal section of an onion root tip. 2)Focus first with the low-power objective 3)Identify cells in interphase, prophase, petaphase, anaphase, and telophase 4)Look at 40-50 onion root tip cells, and tally the frequency of occurrence of the mitotic phases. 2.Staining dividing cells(garlic root tips) 1)Pour 1 ml of freshly prepared C2H6COOH fixative into a small vial. Remove about 2-3 mm from the tips of garlic root and place them immediately in the fixative. The root tips should be fixed for 15 minutes at 60!. 2)After fixation, slowly pour off the fixation into a waster container. 3)Add 1mL 1M HCl to the vial and incubate the vial in a water bath for 10 minutes at 60! 4)After hydrolysis, pour off the acid into a waster container. Add 1mL Feulgen stain to the vial. " /3 1 "

Maximum stain intensity will be reached after 45 mins to 1h at room temperature. 5)Transfer a root tip to a very small drop of 45% C2H6COOH on a slide. Place a clean coverslip over the preparation. 6)Lay the slide on the table and cover the slide with two thicknesses of paper towels.Press the coverslip directly downward. 7)Observe the chromosomes.Identify cells in the four mitotic phases and in interphase.

Results
1.Observe mitosis in plant(cell onion root tip)
Data Table 1 Determining duration of cell cycle Phase Interphase Mitotic phase Total Number Seen 36 15 51 % of Total 70.59% 29.41% 100%

From Data Table 1, we can find most cells are in interphase. Interphase lasts for most of the cell cycle.

2.Staining dividing cells(garlic root tips)

Data Table 2 Determining duration of mitotic phase Phase Prophase Metaphase Anaphase Telophase Number Seen 22 4 2 2 30 % of Total 73.33% 13.33% 6.67% 6.67% 100%

Total

When we observe the slide of garlic root tips which be stained, we can find cells in four mitotic phase and in interphase. When cells are in interphase(Fig.1), the chromosomes cannot be distinguished individually. They are still in the form of loosely packed chromatin fibers. When cells are in prophase(Fig.2), the chromatin fibers coil, so that the chromosomes became thick enough to be seen individually. When cells are in metaphase(Fig.3), we can observe that the centromeres of all the chromosomes line up between the two poles of the spindle, in the middle of the cell. When cells are in anaphase(Fig.4), we can observe that the sister chromatids of each chromosome separate and move toward opposite poles of the cell. When cells are in telophase(Fig.5), we can observe that the two groups of chromosomes have reached opposite ends of the cell. And nuclear envelops start forming. " /3 2 "

Discussion
Question 1.What is the function of mitosis in cell cycle? In mitosis, the nucleus and its contents, most importantly the duplicated chromosomes, divided and are evenly distributed, forming two daughter nuclei.The combination of mitosis and cytokinesis produces two genetically identical daughter cells, each with a single nucleus, the surrounding cytoplasm with organelles, and a plasma membrane. 2.Besides Feulgen stain, suggest other methods to visualize chromosomes in mitotic cells? Using diphenylamine. After hydrolysis, pour off the acid and add 4mL diphenylamine. Heat the vial in the water bath 100 ! for 5 mins. The DNA will be dyed to blue. Using Methyl Green. After hydrolysis, pour off the acid. Add some drop of 1% NaHCO3 into the vial to neutralize the H+. Repeat 3 times. Then add some drops of Methyl Green. The DNA will be dyed to green. At the first time we made the slide of garlic root tips, the coverslip moved sideways. When we observed the slide, we found cells separated with each other. There are few cells in the field of vision. Thats because moving coverslip sideways rolls cells on top of one another and destroy the material. And too much water remained, which made the cells flow everywhere. So its important to press the coverslip directly downward and quickly, and use paper towel to absorb the liquid as much as possible before press the coverslip.

References
Campbell Essential Biology wikipedia.com Lab manual baike.badu.com

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Contribution statements
We do this experience together.

Figure

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Fig.1 Cells of garlic root tips in interphase /10*40 Fig.2 Cells of garlic root tips in prophase/10*40 Fig.3 Cells of garlic root tips in metaphase/10*40 Fig.4 Cells of garlic root tips in anaphase/10*40 Fig.5 Cells of garlic root tips in telophase/10*40 " /3 3 "