Name__________________ Biology 211 Intro Molecular and Cell Biology Mitosis Laboratory Purpose: To prepare slides of onion root tips demonstrating the stages of mitosis (somatic cell division) and observe various cell types undergoing mitosis. Reading: Campbell et al., Chap. 12 Background: Mitosis is the process of cell division that occurs in somatic (body) cells. In mitosis, a cell divides to give two daughter cells, essentially identical to the parent cell. Mitosis results in an equal distribution of hereditary material and usually an equal distribution of the cell contents. All of us began life as single cells. These cells divided by mitosis to become 2, then 4, then 8, then 16 cells and so on. At some point some of these cells started to differentiate into other cell types to form our various tissues. In cancer, the process of cell division has gone awry. To begin to understand the kinds of changes that occur that allow cells to grow and divide past their normal "checkpoints", it is crucial to understand the process of division in normal cells. Materials: onion root tips watch glasses acetocarmine stain (1% in 45% acetic acid, boiled 5 min. and filtered) 1 M HCl razor blades forceps slide warmer slides, coverslips compound microscopes, lens paper mitosis slides: Allium (onion) root tip, whitefish blastula Procedure: Source: Mertens and Hammersmith (2001) Genetics Laboratory Investigations, twelfth edition. Prentice Hall. 1. Prepare slides of onion root tip. a. Clip the terminal 1 cm of the root tip from a growing onion bulb. 1 b. Place several drops of 1 M HCl in a watch glass. c. Place the terminal 3 mm of the onion root in the HCl solution. d. Incubate at room T for 3-5 min. e. Using a forceps, transfer the root tip to a drop of acetocarmine stain on a clean slide. f. Using a razor blade or sharp scalpel, cut off and retain the tip-most 1 mm of the root. Chop the root tip into many pieces with the razor blade. g. Apply a clean cover glass to the slide and heat it gently for 5 min. on a slide warmer. h. Cover the slide with a paper towel and push downward firmly with your thumb over the cover glass. i. Examine with the light microscope at low power and high power. Look for cells where the nuclei have stained red-purple. Interphase cell Label: Nucleus, nuclear envelope, cell membrane Mitotic cell: For extra credit identify and draw an onion root cell from your slide in a stage of mitosis See instructor or lab assistant for help locating mitotic cells. 2. Review the process of mitosis using the mitosis poster or laminated diagrams. 3. Observe mitosis on prepared slides of onion root tip and whitefish. Identify cells in the stages of interphase, prophase, metaphase, anaphase, and telophase. 2 Interphase: Chromatin appears dispersed, DNA replication occurs. Prophase: Chromatin condenses, chromosomes become visible, nuclear membrane breaks down, spindle starts to form. Metaphase: Chromosomes line up on the spindle in the center of the cell (on metaphase plate). Anaphase: Chromosomes are separated at their centromeres, spindle pulls them toward opposite poles. Telophase: Chromosomes recondense, new cell wall forms between daughter cells (plant cells) or cell membrane pinches off (animal cells). Organism ____________. Draw cells in each stage for either onion or whitefish. Interphase Label: Nucleus, chromatin, nuclear envelope, cytoplasm Prophase Label: Chromosomes, spindle Metaphase Label: Chromosomes, spindle, metaphase plate 3 Anaphase Label: Chromosomes, microtubules Telophase Label: Nuclear membrane, cell membrane, chromosomes 4. Ask yourself the following questions as you study the slides: a. Chromosome division: When do the following changes take place? DNA replication occurs Chromosomes condense Chromosomes disperse Sister chromatids are attached at the centromere Centromere divides and sister chromosomes separate b. Changes in cell structure: When do the following events take place? Breakdown of nuclear envelope Spindle forms 4 Nuclear membrane reforms Cell membrane reforms Cell wall is formed c. What are some of the differences between the process of mitosis as it occurs in plants (onion) and animals (whitefish)? Assignment: (20 points). Complete the drawings and tables above. 5