Mitosis

Lab#2
Why is an onion root tip good for studying
mitosis?

Onion root tips are extremely popular for viewing the

different phases of mitosis because the chromosomes are
large and and very dark when stained. ...
The real size of a typical onion root tip cell is about .075mm
(75 micrometers).
The aim of cell cycle
• To distribute complete and accurate replicas of the
genome to daughter cells 2.
• A. Interphase- is the stage
between two successive
cell divisions.
• It occurs before mitosis
begins.
 Interphase includes 3 phases as follows:

1.Gap 1 (G1) phase-

(1-5 hr.)
-the cell increases in size
-RNA and proteins including
enzymes need for making
DNA are synthesized.
2. Synthesis (S) phase
(5-12hr)
- DNA is synthesized
and
- chromosomes are
replicated.
3. Gap 2 (G2) phases-
(12-15hr)
-continues to synthesize
RNA and proteins
-cell continue to increase
in size.
The checkpoints are surveillance mechanism and
quality control of the genome to maintain genomic
integrity.
DNA replication checkpoint that arrests cell
cycle at G2/M transition until DNA replication
is complete.

•Spindle checkpoint The spindle checkpoint

arrests cell cycle at M phase until all
chromosomes are aligned on spindle. This
checkpoint is very important for equal
distribution of chromosomes.
•Morphogenesis checkpoint detects
abnormality in cytoskeleton and arrests cell
cycle at G2/M transition.
During interphase:
 INTERPHASE
Animal Cell Plant Cell
1.PROPHASE

Centrioles- or poles) begin to move to opposite end of

the cell.
Spindle fibers form between the poles
 METAPHASE
Animal Cell Plant Cell

Photographs from: http://www.bioweb.uncc.edu/biol1110/Stages.htm

 3. ANAPHASE
Chromatid (or pairs
of chromosomes)
separate and
begin to move
towards the
opposite ends of
the cell.
 ANAPHASE
Animal Cell Plant Cell

Photographs from: http://www.bioweb.uncc.edu/biol1110/Stages.htm

4. TELOPHASE
Two new nuclei form.
Mitosis
 Telophase
Animal Cell Plant Cell

Photographs from: http://www.bioweb.uncc.edu/biol1110/Stages.htm

Harvesting and Fixing the Root Tips
Obtain test tube, pair of scissors, watch glass, and fixative (9 parts 45% acetic acid and 1 part HCL).
Cut off four root tips, each approximately 1 cm long; Fill test tube 3cm full of fixative.
Place four root tips into the test tube of fixative and incubate at
50 degrees Celsius for six minutes. Then dump heated fixative
and tips into watch glass.
Cut all excess from the root tips except for 2mm at the very tip
of the root (end that was not cut from the plant).
Place two drops of aceto-orcein stain on top of the 2mm root
tip.
Let stain soak into root tip for two minutes.
Squash the root tip, on each slide, pressing straight down so as
not to overlap the cells.
Place two more drops of stain upon the root tip and wait for another two
minutes.
Then place cover slip flat upon the root tip, making certain not to move
the cover slip horizontally.
Press the cover slip gently with a pencil eraser, again only straight down
without moving the cover slip.
Soak up extra stain from slide around cover slip with a paper towel
without moving cover slip.
Observe and record steps of cell division under microscope (at 400x)
Sample views at 400x: