Microscopy

Optical Microscopy
Oldest and simplest of microscope,
using visible light and lenses.
Basic configuration single and
compound lens
Reflected or transmitted light is used as
source
Magnification between 10 1400x
Basically any types of sample can be
observed

Metal

optical microscope view of a crosssection through the baked

comparison
tube
Source:
met-tech.com/images/steel-heater-tools

Polymer

Optical Microscope image of buckled

PMMA film over patterned PDMS
holes.
Source: www.pse.umass.edu/acrosby/YuriEbata.htm

Ceramic matrix Composite

Mat. Res.vol.10no.2So CarlosApr./June2007

Source: www.scielo.br/scielo.php?pid=S1516-1439200700...

Semiconductor Packaging

Source: www.caltexsci.com/CX-3000.htm

Basic configurations
1 .ocular lens, or eyepiece
2 .objective turret
3 objective lenses
4 coarse adjustment knob
5 fine adjustment knob
6 object holder or stage
7 mirror or light (illuminator)
8.diaphragm and condenser
Source: wikipedia.

Image formation

Resolution
The closest spacing of two points
which can be clearly be seen through
microscope to be separate entities
(Goodhew et al , Electron Microscopy and Analysis, 3rd Edition)

Resolution limited by diffraction

effect
Whenever light passes through
aperture, diffraction occurs

Airy rings

R1 =d1/2 = 0.61/ sin , = refractive index, = semi angle

sin = NA, numerical aperture
Good microscope can have resolution up to 150 nm using green
light ( =400 -550nm)
sin = nearly unity using an oil immersion objective lens

 Usually, an of 550 nm is assumed,

corresponding to green light. With air
as medium, the highest practical NA
is 0.95, and with oil, up to 1.5. In
practice the lowest value of d
obtainable is about 200 nm.

Depth of field
Measure of how much of the object we are
looking remains in focus at the same time.
(governed by the lenses in the microscope)
Image is accurately in focus when the object lies
in the approriate plane (e.g surface of sphere)
h= 0.61/ sin tan
Increase h by decreasing
convergence angle

Aberrations in optical
system
Lens aberration- chromatic and
monochromatic aberrations
Improve chromatic abr.- combining
lenses with diff refractive indices or
using filter at source
Improve monochromatic abr.-reduced
only the central portion of lens are
used.

Chromatic Abrr.

Monochromatic Abr

astigmatism

Variants of Microscope
Differential interference contrast
microscopy (DIC), or Nomarski microscopy
Phase contrast microscopy- e.g study cell
cycle
Fluorescence microscope
Inverted microscope
Petrographic microscope/polarized
miccroscope-e.g detailed descriptions of
rocks

Improved resolution
microscopy
Vertico SMI (Spatially Modulated
Illumination)
Up to 10nm

Near-field scanning optical microscopy

(NSOM/SNOM)
lateral resolution of 20 nm and vertical
resolution of 25 nm

Stimulated Emission Depletion microscopy,

or STED microscopy
Up to 5.8 nm can be achieved