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CGP Online Edition

36 37

ICAL
PRACT S
SKILL Cell
Cell Division
Division —
—Investigating
InvestigatingMitosis
Mitosis PRACT
ICA
SKILL L
S

It’s time to dust off your lab coat and get out your safety specs. Here are all the techniques you need to study mitosis.
Youneed
You’ll CantoUseknowAhow Graticule
to stain rootand Micrometer
cells on touse
slides and how to Calculate the Size and
an optical microscope of Cells ...
graticules.

1 ) You need to be able to calculate the size of the cells you’re looking at.
Root Tips Can be Stained and Squashed to Observe Mitosis
That’s where the eyepiece graticule and stage micrometer come in — they’re a bit like rulers.
You
2) need
An eyepiece how to prepare
to know graticule is fittedand
ontostain root tip. inIt’sorder
the aeyepiece like to stages
observe theruler
a transparent of numbers
with mitosis. ,Make
but nosure
units.
you’re
3) The stage safety
wearing goggles
micrometer is and
placeda lab
oncoat before—you
the stage it isstart. You should
a microscope alsowith
slide wearangloves when
accurate using
scale stains
(it has .
units)
and it’s used to work out the value of the divisions on the eyepiece graticule at a particular magnification.
1 ) Cut 1 cm from the tip from a growing root (e.g. of an onion). It needs to be the tip
4) This means
because thatwhere
that’s when growth
you takeoccurs
the stage
(andmicrometer
so that’s where awaymitosis
and replace it with the slide containing your
takes place).
tissue sample,
If you’re you’ll be able
using ethano-orcein to measure
to stain the
the cells, the tipssize
will of
alsothe cells.
need to beHere’s
fixed in an example
ethanoic acid.:
2) Prepare a boiling tube containing 1 M hydrochloric 1 ) Lineacidupand
the put
eyepiece a water and
it in graticule baththeatstage 60 ˚Cmicrometer.
.
3) Transfer
4.5 the root tip into Eyepiece graticuletube and incubate
the boiling 2) Each for abouton5 the
division minutes .
stage micrometer is 0.1 mm long.
4) Useeyepiece
a pipette to rinse the root tip well with cold
0 1 0 20 30 40 50
3) water . Leave
At this the tip to
magnification, dry on on
1 division a paper
the stage towel .
micrometer is the
divisions = same as 4.5 divisions on the eyepiece graticule .
5) Place the root tip on a microscope slide and cut 2 mm from the very tip of it. Get rid of the rest.
1 stage
a mounted 4) spread
To work theoutcells
the outsize of 1 division on the eyepiece graticule,
6) Usedivision 1 0 needle 20 to break the 30 tip open and thinly.
Stainedwith RootFeulgen's
Cells
you need to divide 0.1 by Root 4.5: cells stained reagent
7) Add a few drops of stain and leave it for a few minutes. 1 division on eyepiece graticule = 0.1 ÷ 4.5 = 0.022 mm
The stain will make the chromosomes easier to see under a
1 division = 0.1 mm 5) So if you lookTelophaseat a cell under the microscope at this magnification
microscope. There are loads of different stains, all with crazy
and it’s 4 eyepiece divisions long, you know it measures:
names (toluidine blue Stage O , ethano-orcein, Feulgen stain
micrometer 4 × ... 0.088 mm. | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | |
0.022 =Prophase

| | | | | | ||
If you’re using the Feulgen stain, you’ll need an extra rinse. The eyepiece graticule will need to be

| | | | | ||
Anaphase re-ca librated at different mag nifications.
8) Place a cover slip over the cells and push down firmly to | | | | | | | | | | | | | | | | | | | |
| | | | | | | | | | | | | | | |

...Or
squashYouthe Can Use This
tissue. Thiswill Formula
make the ...tissue thinner and Metaphase
allow light to pass through it. Don’t smear the cover slip
If you’re
sideways given(oranyou’llimage
damageof cellstheunder the microscope in the exam,
chromosomes). Interphase
size of image
you can calculate their actual size using this formula: actual size
H E RV E= C O N G E , I S M / S C I E N C E P H O T O L I B R A RY
9) Now you can look at all the stages of mitosis under magnification
|| | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | |
Example:
an optical If the image of a cell
microscope (seemeasures
below). 5 mm
Youand the magnification
should see is | | | | ||
| | | | | | |

You need to be able to recognise cells in the different

| | | | | | |
x something
1 00 , then the that actual sizelike
looks of the
thecell will be: 5 ÷ 1on
photograph 00the= 0.05
right.mm. stages of mitosis — see p. 34 for more info.
| | | | | | ||
|| | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | |

Artefacts Can Get in the Way of Your Observations

You Can Observe Cells Using an Optical Microscope
1 ) Artefacts are things that you can see down the microscope that
You need to know how to use an optical microscope to observe your prepared root tip cells: If you’re asked to draw
aren’t part of the cell or specimen that you’re looking at.
cells undergoing mitosis
12)) Start
They by
canclipping
be anything fromyou’ve
the slide dust, air onto
bits ofprepared bubbles stagefingerprints
the and . , Theunder the microscope,
new organelle Steve
to inaccuracies caused by sobjective
quashing and staining your sample. Eyepiece
2) Select the lowest-powered lens make sure looked
had discovered you write
j ust
3) (i.e. the one
Artefacts are that produces
usually made the lowest
during the magnification).
preparation of your slides and shouldn’t really be downhisthe
like magnification
thumb print.
Coarse the specimen was viewed
therethe
3) Use at all — you’ll
coarse need toknob
adjustment prepare yourthe
to bring rootstage up tocarefully
tip cells just to avoid creating artefacts.
adj ustment under. You’ll also need
below the objective lens. knob to label your drawing.
Artefacts are especially common in electron micrographs because specimens need a lot of
4) Look down the eyepiece (which contains the ocular lens).
preparation before you can view them under an electron microscope. The first scientists to
Use the coarse adjustment knob to move the stage downwards,
use these microscopes could only distinguish between artefacts and organelles by repeatedly High and low power
away from the objective lens until the image is roughly in focus. Fine with one preparation
preparing specimens in different ways. If an object could be seen obj ective lenses
the focus with
5) Adjust technique, the fine adjustment knob , until adj ustment
but not another, it was more likely to be an artefact than an organelle.
you get a clear image of what’s on the slide. knob Stage
6) If you need to see the slide with greater magnification, swap to Light
Warm-Up Questions
a higher-powered objective lens and refocus. P RA
QU E C T I C E
STI
Q1 Why do you need to squash the tissue when preparing a slide of plant root tip cells? ON
S

The Mitotic
Exam Index Is the Proportion of Cells Undergoing Mitosis
Question
You can
Q1 calculate
A sample of cells mitotic
thewas index
prepared to of your cells
observe using
mitosis. total,formula
In this 42 cells: were observed. 32 of those had visible
chromosomes. Calculate the mitotic index for this sample. Give your answer to 2 decimal places. [2 marks]
mitotic index = number of cells with visible chromosomes
total number of cells observed
‘Staining your samples’ — a common problem at the start of exams...
This lets you work out how quickly the tissue is growing and if there’s anything weird going on. A plant root tip is
Wconstantly
ow — I betgrowing
you never
, sorealised there awas
you’d expect so mitotic
high much toindex
know(i.e.
about
lotsusing a microscope.
of cells Still,
in mitosis). In staining
other tissueissamples,
pretty
straightforward
a high mitoticand so’scould
index preparing
mean athat
slide. Using
tissue a graticule
repair is takingisplace
tricky,orbut once
that you
there get your head
is cancerous round
growth in it
theyou’ll be fine.
tissue.
Topic 2A — Cell Structure and Division Topic 2A — Cell Structure and Division

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