Phase Contrast

Microscope
 PHASE CONTRAST MICROSCOPE
 In 1934 Frits Zernike, a Dutch Physicist
created an optical design that could
transform differences in phase to
differences of amplitude.
 This phase contrast technique proved
to be such an advancement in
microscopy that Zernike was awarded
Nobel prize (physics) in 1953.
 OPTICAL FUNDAMENTALS
 Lightpassing through different parts of the glass window
would actually travel through different distances of glass
depending on the glass thickness at a particular point, as
shown in the diagram
 The peaks and troughs of the waves no longer match
up
 We say these waves have a difference in 'phase‘
 phase differences arise because the light travels
through different distances depending on the thickness of
the glass at each point.
 a phase contrast microscope is used to transform phase
differences into intensity differences, increasing contrast.
CONDENSER ANNULUS
(annular diaphragm)
situated below the
condenser
Made up of opaque flat
black plate with
transparent circular
disc having circular
annular groove
 PHASE PLATE
A transparent disc placed in
back focal plane of objective.
*negative phase plate
*positive phase plate
The portion of the phase plate upon which
the condenser annulus is focused is termed
the conjugate area, while the remaining
regions are collectively referred to as
the complementary area.