1. Mitotic Cell Division

2. Introduction
The growth and development of every organism depends on the precise replication of the genetic material during each cell division.
Cell division is the process by which cells reproduce themselves.

3. Objectives
Learn preparing and staining procedure to identify the stages of mitosis in onion root tip.
To differentiate between the different stages of mitosis.

4. Types of Cell Division Mitosis
Mitosis is the process by which a eukaryotic cell separates the chromosomes in its cell nucleus into two identical sets, in two separate nuclei.
It is generally followed immediately by cytokinesis,
The outcome of this process: two new daughter cells with the same number and kind of chromosomes as the parent cell. .

5. New body (somatic) cells are formed by mitosis.
Meiosis ( Reduction division )
produces progeny cells with one-half the genetic content and number of chromosomes as parent cell
The formation of male and female gametes in animal cells or spores in plant cells is by meiosis.

6. Mitosis
Purpose
Mitosis occurs in order for organisms to grow and develop.
In order to replenish dead or dying cells such as skin cells, and cells in the digestive tract.
Karyokinesis
process of nuclear division (division of genetic material).
Cytokinesis
Process of dividing cytoplasm/cell.

7. Mitosis occurs only in eukaryotic cells and the process varies in different species. For example:
animals undergo an "open" mitosis, where the nuclear envelope breaks down before the chromosomes separate.
Fungi and yeast undergo a "closed" mitosis, where chromosomes divide within an intact cell nucleus.
Prokaryotic cells, which lack a nucleus, divide by a process called binary fission.

8. Errors in mitosis can either kill a cell through apoptosis or cause mutations that may lead to certain types of cancer.

9. The Cell Cycle
The life of a cell is divided into three stages known as the cell cycle:
1. Interphase: cell carries out normal functions and prepares to divide.
2. Mitosis: nucleus divides splits into two.
3. Cytokinesis: cell and contents divide into two daughter cells.

10. Interphase The cell prepares itself for cell division.
This phase consist of the G1 (first gap), S (synthesis), and G2 (second gap) phases.
The chromatin is diffuse.
protein synthesis,
DNA synthesis,
Replication of other cellular structures.

11. S phase: where each chromosome is duplicated and consists of two sister chromatids joined together by a centromere.
Now, the nucleus and cell increase in
size, and chromosomes are fully extended.

12. Mitosis There are 4 main phases: Prophase, Metaphase, Anaphase,
Telophase.
Cytokinesis (division of the cytoplasm) follows and one cell becomes two.
the nucleus has to migrate into the center of the cell before mitosis can begin.

13. Mitosis: Prophase Major processes during this phase:
Chromosomes condense and form visible bodies.
Chromosomes become thicker, shorter, and easily visible when stained under the light microscope.
Two “sister chromatids” join near their middle at a structure called the centromere.
The nucleolus and the nuclear membrane disappear.
The mitotic apparatus the spindle, begins to organize within the cell.
Each chromosome forms two kinetochores at its centromere, one attached at each chromatid.
A kinetochore is a complex protein structure and it is the point where microtubules attach themselves to the chromosome

14. Mitosis: Metaphase
Chromosomes become aligned at midpoint or equator between poles of the cell.
Are at their thickest and shortest structure.
They are easily identified as two longitudinally double sister chromatids.
Chromatids are connected (at their centromeres) to the spindle apparatus, which has formed between the two centrioles located at the poles of the cell.

15. Mitosis: Anaphase The centromere replicates and splits
The sister chromatids begin to separate and migrate to the opposite poles separate of the cell.
It "cleans up" the after effects of mitosis
For humans, with a diploid number of 46 chromosomes, there will be 46 chromosomes moving toward each pole.
Onions have 16 diploid chromosomes and, therefore 16 chromosomes move to each pole.

16. Mitosis: Telophase Chromosomes now uncoil
Nuclear envelope reappears and surrounds the chromosomes
Cytokinesis
The cytoplasm and all its contents are divided between the 2 daughter cells (cytoplasmic division), membrane creates between the 2 new daughter cells
In plants, such as the onion root tip cells, this is seen as the formation of a cell plate
A new nuclear membrane, using the membrane vesicles of the parent cell's old nuclear membrane, forms around each set of separated daughter chromosomes.
Cytokinesis is technically not even a phase of mitosis, but rather a separate process, necessary for completing cell division

17. Cell Cycle

19. Stages of mitosis in onion root tip cells

20. Mitosis in Root Tip
In a growing plant root, the cells at the tip of the root are constantly dividing to allow the root to grow.
Because each cell divides independently of the others, a root tip contains cells at different stages of the cell cycle.
This makes a root tip an excellent tissue to study the stages of cell division.

24. Experiment

25. Materials Slides & cover slips Microscope Fresh onion root tips
Fixative ( methanol-acetic acid 3:1 v/v)
Forceps
1 M HCl
Razor blade
Stain
Paper towel, or absorbent paper

26. Method Cut 2-3 mm of onion root
Use forceps to transfer an onion root tip into the cup of HCl.
Leave for 4 minutes
Transfer the root tip to the cup containing fixative and leave it for 4 minutes.
Then place the root tip on a slide.
Cover the root tip with a few drops of stain for 2 minutes
Cover the root tip with one to two drops of 45% acetic acid
Put a cover slip over the root, put a paper towel or other absorbent paper and with your thumb firmly press on the cover slip.

27. Observe your preparation under the low power (X10) of a microscope
Search the slide to find cells in various stages of cell division, once you have located cells in division, change to high power (X40) & try to observe several stages of division.
Record the number of cells in each stage. Count at least three full fields of view. You should have counted over 200 cells.
Record your data in the table
Calculate the percentage of cells in each phase and record in the table

28. Interphase
Prophase
Anaphase
Metaphase
Telophase

29. Animation http://www.youtube.com/watch?v=s1ylUTbXyWU