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Required Practical 2 Bio

The document outlines a practical experiment to observe mitosis in root tips and calculate the mitotic index using an optical microscope and various reagents. It details the equipment needed, the step-by-step method for preparing and observing the samples, and includes a risk assessment for handling hazardous materials. The objective is to count the number of cells in mitosis to determine the mitotic index, which is the ratio of cells undergoing mitosis to the total number of cells observed.

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30 views

Required Practical 2 Bio

The document outlines a practical experiment to observe mitosis in root tips and calculate the mitotic index using an optical microscope and various reagents. It details the equipment needed, the step-by-step method for preparing and observing the samples, and includes a risk assessment for handling hazardous materials. The objective is to count the number of cells in mitosis to determine the mitotic index, which is the ratio of cells undergoing mitosis to the total number of cells observed.

Uploaded by

gurnoor.pawra
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as DOCX, PDF, TXT or read online on Scribd
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Required Practical 2: Observing mitosis

Mitosis is a type of cell division for cell repair and growth, it produces two genetically identical
daughter cells. The objective of this practical is to count the number of cells in mitosis to
calculate the mitotic index.

Mitotic Index = (number of cells in mitosis)/ total number of cells

Equipment:

 Optical microscope
 Microscope slides
 Cover slips
 Water bath
 Hydrochloric acid (5 mol dm-3)
 Toluidine blue O stain
 Distilled water
 Scalpel & forceps
 100 ml beaker
 Root tip
 Stopwatch
 Watch glass

Method:

1. Cut 2cm of root tips – root tip is cut as mitosis occurs at the meristem and the
objective of this practical is to observe cell division.
2. Prepare water bath containing hydrochloric acid and place root tips into beaker
containing solution – the root tip is heated with acid to macerate cells and break cell
walls.
3. Start stopwatch and leave root tip in heated hydrochloric acid solution for 10 minutes.
4. After 10 minutes, rinse root tip in distilled water in the watch glass.
5. Place root tip on slide and add stain – Toluidine blue O stains tissues rich in DNA and
RNA, therefore making processes in mitosis visible.
6. Place cover slip and apply pressure – this is done so cells can be squashed into a single
layer of cells to allow light to pass through. Do not rub the slide.
7. Look through eyepiece lens and use the lowest objective lens first. Use the coarse and
fine focusing wheel to focus image.
8. Record the number of cells visible and the number of cells taking place in mitosis, in a
table of values.
9. Repeat the same process for several fields of view.
10. Calculate the mitotic index based on the data recorded.
Hazard Risk Precaution Risk Level
Hydrochloric acid Corrosive Use of PPE, ensure goggles are worn to Low
prevent any splashes of the acid into the eye.
In case of skin contact, rinse affected area
with water immediately. Spillages should be
neutralized using sodium bicarbonate to
prevent any further damage.

Sharp tools (e.g Cuts, Ensure the direction you are cutting in is Low
scalpel) scratches facing away from the body. Keep scalpel away
(can allow from edge of surfaces and hold them pointing
bacteria into downwards when collecting equipment. Do
the not leave any sharp objects unattended.
bloodstream) If injury does occur, rinse minor scratched and
cuts in cold water. For any severe injuries,
elevate cuts and apply pressure and let your
teacher know.
Toluidine blue O stain May cause Avoid contact with skin and wear eye Low
irritation to protection e.g goggles. In case skin comes in
skin. contact, wash the affected area immediately
and rinse with cold water.

Risk Assessment:
References:

AQA, n.d. A-level Biology: Paper 2. Available at:


https://filestore.aqa.org.uk/resources/biology/AQA-7401-7402-P2.PDF [Accessed 29
March 2025].

Save My Exams, n.d. Recognising the Stages of Mitosis. Available at:


https://www.savemyexams.com/a-level/biology/aqa/17/revision-notes/2-cell-structure/2-
3-cell-division-in-eukaryotic-and-prokaryotic-cells/2-3-5-recognising-the-stages-of-mitosis/
[Accessed 29 March 2025].

National Center for Biotechnology Information (NCBI), 2025. Hydrochloric Acid. PubChem.
Available at: https://pubchem.ncbi.nlm.nih.gov/compound/Hydrochloric-Acid [Accessed
29 March 2025].

University of Manchester, n.d. Sharps General Risk Assessment. Available at:


https://www.staffnet.manchester.ac.uk/media/eps/chemistry-intranet/health-and-
safety/Sharps_General_Risk_Assessment.doc [Accessed 29 March 2025].

Edvotek, n.d. Toluidine Blue Stain O. Available at:


https://www.edvotek.com/site/pdf/Toluidine%20Blue%20Stain%20O.pdf [Accessed 29
March 2025].

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