Unit 1 Revision Microscopy

Total magnification = Magnification of Eye Piece lens x Magnification of Objective lens

The eyepiece lens usually has a x10 magnification and the greatest magnification of the objective lens is usually x100, so
therefore the greatest total magnification is usually x1000.

Magnification is the degree to which the object is enlarged.

Numerically, it is the image size divided by the actual size of the
object, measured using the same units. It is usually expressed as x10,
x1.5, etc.

Resolution is the degree to which it is possible to distinguish between

two objects that are very close together. The higher the resolution,
the greater the detail you can see.

The Specimen

The material to be viewed on the slide or specimen needs to be thin,

so that light or an electron beam can pass through it. A coverslip is needed to protect the specimen and the lens if they
where to touch.

Stains

Stains can be used in order to distinguish different features present in the specimen.

methyl-blue  check cells

iodine  plant cells

acetic orcein  nuclei and chromosomes

Differential staining

Used to distinguish between two types of organisms that would otherwise be difficult to identify.

It can also be used to differentiate between different organelles of a single organism within a tissue sample.

How to use a microscope

1.Lower the stage and mount the slide using the clips.

2.Make sure the objective lens is set to the lowest magnification one.

3.Carefully turn the coarse focusing know to bring the stage up to the top.

4.Looking through the microscope slowly lower the stage (using the coarse
focusing knob) until you can see the specimen. You might need to move the
slide to get it nice and centre.

5.Use the fine focusing knob to bring the specimen into sharp focus.

6.Change the magnification by the choosing the next objective lens up, you
should only have to change the focus (on the fine focusing knob) and not touch
the coarse focusing knob.

Calibration of Microscope

1. Line up the eyepiece graticule and the stage micrometer.

2. Each division of the stage micrometer is 0.1mm long.
3. At this magnification 1 division on the stage micrometer is equal to 4.5 divisions on the eyepiece graticule.
4. To work out the size of one eyepiece graticule division you divide the length of one stage micrometer division by the
number of eyepiece graticule divisions.
5. So, if you were to look at an object under the microscope at this magnification you times the number of divisions long
it is by 0.022.