Cell Cycle Lab

Examining Mitosis in the root tip of Allium Already prepared on microscope slides Objective to observe and identify Interphase,

Prophase, Metaphase, Anaphase, and Telophase

Mitosis Phases
Interphase. The cell is engaged in metabolic activity and performing its duty as part of a tissue. The DNA duplicates during interphase to prepare for mitosis (the next four phases that lead up to and include nuclear division). Chromosomes are not clearly discerned in the nucleus, although a dark spot called the nucleolus may be visible. Prophase. Chromatin in the nucleus begins to condense and becomes visible in the light microscope as chromosomes. The nuclear membrane dissolves, marking the beginning of prometaphase. Proteins attach to the centromeres creating the kinetochores. Microtubules attach at the kinetochores and the chromosomes begin moving. Metaphase. Spindle fibers align the chromosomes along the middle of the cell nucleus. This line is referred to as the metaphase plate. This organization helps to ensure that in the next phase, when the chromosomes are separated, each new nucleus will receive one copy of each chromosome. Anaphase. The paired chromosomes separate at the kinetochores and move to opposite sides of the cell. Motion results from a combination of kinetochore movement along the spindle microtubules and through the physical interaction of polar microtubules. Telophase. New membranes form around the daughter nuclei while the chromosomes disperse and are no longer visible under the light microscope. Cytokinesis or the partitioning of the cell may also begin during this stage.

Lab
Groups of 2 1 microscope per group 1 microscope slide per group View slide at 40X Count 100 cells One group member will observe and one will mark done observations for 50 cells. Then switch and observe remaining 50 cells on a different part of the slide.

1 interphase 2 interphase 3 early prophase 4 mid prophase 5 late prophase 6 metaphase 7 early anaphase 8 anaphase 9 early telophase 10 telophase

Microscope Usage
Handle microscope with 2 hands One hand on the base and one hand holding arm of scope Focus at 4X first Only time you use course focus (large knob) Then focus at 10X using fine focus (small knob) Switch to 40X and focus using fine focus

Lab Make table of your small groups observations: Interphase

Prophase Metaphase Anaphase Telophase

20 20 20 20 20

Pool class data into table

Fill in excel document in class

Meta Telo Observer Interp Prop phas Anap phas To (Group) hase hase e hase e tal Group 1 0 Group 2 0 Group 3 0 Group 4 0 Group 5 0 Group 6 0 TOTAL 0 0 0 0 0 0

Lab Report
Introduction: Background about cell cycle

What are the stages and what is happening in each stage Relative amount of time spent in each stage

Hypothesis

Lab Report
Methods: Prepared slides Count cells/ determine stage Discuss calculations

Lab Report
Results:
Include

table of class data for each phase Calculate mean, S.D., and S.E. of CLASS DATA for each phase Generate table of small group results
Interphase Prophase Metaphase Anaphase Telophase 20 20 20 20 20

Lab Report
Calculate amount of time cells spent in each stage

of mitosis.
Example: Calculate percentage of time spent in a specific stage from class data

Interphase: 756/1200 X 100 = 63% 16 hours (time it takes to complete cell cycle) x 0.63 = 10.08 hours 0.08 hours x 60 minutes/1 hour = 4.8 minutes 0.8 minutes x 60 seconds/1 minute = 48 seconds Answer = 10 hours 4 mins 48 seconds in interphase

Use percentage to calculate time spent in stage

Lab Report
Calculate variability measurement Used to measure

variability of class in determination of stages

Higher number indicates more variability

Use class data to calculate variability Calculate for each stage Variability = S.E./Mean

Lab Report
Results Make pie chart of the percentage of time spent in each stage Use class data for chart
Telophase, 6% Anaphase, 9%

Metaphase, 7%

Prophase, 15% Interphase, 63%

Lab Report
Discussion Meaning of lab
Why might certain phases take longer, etc. Importance of cell cycle

Lab Report
Conclusion Hypothesis supported? Why/ why not

Lab Report

Introduction ___ Describe cell cycle ___ Discuss cell cycle Methods ___ Counting ___ What the numbers tell us Results ___ Class Data and Calculations (1.5 pts) ___ Individual Data (0.5 pt) ___ Pie Chart ___ Conversion from percent to time Discussion ___ Discuss meaning of the lab Conclusion ___ Hypotheses