Phase contrast is a light microscopy technique used to enhance the contrast of images of

transparent and colourless specimens. It enables visualisation of cells and cell components that would
be difficult to see using an ordinary light microscope.

The specimen is illuminated with a ring of light coming from the annular diaphragm.

Two things happens when light passes thru specimen:

Phase shift depending on the thickness of specimen.

Refraction in different direction because of uneven surface.

- if the refracted light hits the phase ring in the objective, the phase shift of the light increases
and that region of the specimen is seen as dark.

- but if the refracted light fails to hit the face ring then that region of the specimen is seen as
bright.

PHASE CONTRAST MICROSCOPY – can give us very crisp clear details of the overall structure and
shape of the specimen that we are looking under the microscope.

- it is used to enhance contrast to study unstained specimens.

- it allows us to view specimens without staining them, so we can see transparent samples
without staining them.

Annular ring is like a filter – its surrounding portion is totally black and made up of materials
which will not allow any light to pass. It also has slits which allows the light to pass.