MISCROSCOPE BASICS  17th Century

o compound microscope system was

 THE MICROSCOPE (History of Microscope) invented
 Ancient times  incorporates more than one lens
o man has wanted to see things far - image magnified by one lens
smaller than could be perceived with can be further magnified by
the naked eye another
 16th century – led to the  since its invention, it has made
construction of a magnifier tremendous contributions to the
composed of a single convex lens progress of science
 in turn, led to the eventual  Robert Hooke (17th Century)
development of microscope - using a compound
 Most Famous Early Pioneers microscope, he discovered
o Digges of England the fact that living things are
o Hans and Zacharias Janssen of composed of cells
Holland  In the medical world
 Anton van Leeuwenhoek o Louis Pasteur of France
o became the first man to make and  used compound microscope to
use a real microscope discover yeast fungus
o grounded and polished a small glass o Karl Joseph Eberth
ball into a lens with a magnification  German bacteriologist
of 270X  employed a compound
 used it to make the world’s first microscope in his discovery of
practical microscope eberthella typhosa
 had only one lens; referred to as  Robert Koch
a single-lens microscope o used a compound microscope to
 convex glass lens – attached to a discover tubercle and cholera bacilli
metal holder; focused using th
 19 Century
screws o saw dramatic progress in the
o after his historic invention development of microscope
 continued to devote himself to o contributions:
studies based on the microscope  Carl Zeiss – devoted significant
 his discovers included effort to the manufacture of
- bacteria microscope
- animalcules  Ernst Abbe – carried out a
- spermatozoa theoretical study of optical
o constructed a total of 400 principles
microscopes during his prolific  Otto Schott – conducted
lifetime research on optical glass
 THE MICROSCOPE  stage
 Etymology - square platform with
o micro (Latin) – small opening at center
o skopos (Greek) – to look at - where slide is placed during
focusing
 stage clips
- paired structures on either
side of stage
- used to hold slide in place
 coarse adjustment knob
- bigger wheels used to adjust
low power objective in
focusing
 fine adjustment knob
- smaller wheels used for
delicate focusing in
 Parts of the Microscope connection with high-power
A. Mechanical Parts (Framework) objective and oil immersion
 body tube - used to make specimen more
- cylindrical structure arising clear and vivid
vertically from handle  inclination screw
- holds dust shield and - found at junction of pillar
nosepiece with objectives at and handle
lower end - used to tilt microscope
 draw tube  pillar
- upper and smaller end of - short piece of metal attached
body tube bearing eyepiece to one end of base
or ocular - supports microscope
 revolving nosepiece  base
- circular structure at lower - may be Y or U-shaped stand
end of body tube to which - supports microscope
objectives are attached B. Illuminating Parts (Light Control
 dust shield System)
- thin circular structure at  mirror
lower end of body tube used - found below stage, near base
to protect objectives and - provided with concave and
specimen from dust plane surfaces
 arm / handle - used to collect and direct
- curved metallic part arising light to specimen
from pillar  Abbe condenser
- used in holding microscope - lens found immediately
beneath hole of stage
 - used to concentrate light  special oil is placed on object
rays on specimen being studied
 iris diaphragm  has highest degree of
- found below stage consisting magnification
of regularly arranged circular  Kinds of Microscope
blades o Simple / Single
- used to regulate a central  magnifies an object with a single
opening to decrease or lens or enlarges an object solely
increase light reflected on through angular magnification
object  provides viewer an erect
C. Magnifying Parts enlarged virtual image
 ocular / eyepiece  use a single convex lens or
- found on draw tube through groups of lenses
which operator peeps during  ex: magnifying glasses, loupes,
actual focusing and eyepieces for telescopes and
 objectives microscopes
- tube-like structures attached o Compound
to revolving nosepiece  light illuminated
 Types of Objectives  image seen is two-dimensional
o Low-power Objective (LPO)  most commonly used
 shortest tube marked 10X or 6X  can view individual cells, even
affording lowest ocular living ones
magnification o Stereoscopic
 usually shorter than other  best-suited for observation in
objectives low-to-middle magnification
 forms general outline or wider range (approx. 5X – 120X)
portion of object  zoom lens is generally used
o Medium-power Objective (MPO)  camera can be mounted
 longer tube usually marked 30X  advantages
o High-power Objective (HPO) - provide long observation
 lower marked either 40X, 43X, distance
45X - generally employs a zoom
 longer than LPO lens, making it easy to
 forms bigger image of object in determine an observation
focus point
 used to examine / enlarge  disadvantages
specimens that are so small - require additional light
under LPO source
o Oil Immersion Objective (OIO) - effective for observation
 affording a much higher under relatively low
magnification being marked 90- magnification only
100X - most cannot take photos
 - if it can take photos, it is  Transmission Electron
difficult to set up shooting Microscope (TEM)
conditions - first type of EM to be
o Electron developed
 scientific instruments that use a - developed by Max Knoll and
beam of highly energetic Ernst Ruska (Germany, 1931)
electrons to examine objects on - produces image that is a
a very fine scale projection of entire object
 examination can yield the ff (including surface and
information: internal structures)
- topography - incoming electron beam
 surface features of an interacts with the sample as
object or how it looks it passes through the entire
 texture thickness of it
 direct relation between - objects with different
these features and internal structures can be
materials properties differentiated because they
(hardness, reflectivity, give different projections
etc.)  Scanning Electron Microscope
- morphology (SEM)
 shape and size of - debuted in 1942 with first
particles making up the commercial instruments
object around 1965
 direct relation between - when used, bulk biological
structures and materials samples are first coated with
properties (ductility, a metal that readily reflects
strength, reactivity, etc.) electrons
- composition - coating provides conducting
 elements and compounds surface for electrons to avoid
that the object is charging of sample; incoming
composed of and their electron beam is condensed
relative amount into a small beam – scanned
 direct relationship over the object
between composition - image is formed by electrons
and materials properties that bounce off the surface
(melting point, reactivity, of specimen; collected onto
hardness, etc.) the imaging screen
- crystallographic information - observer sees picture of
 how atoms are arranged surface of sample without
in the object any internal information
 o Digital o never leave it in a tilted or inclined
 uses optics and a charge-coupled position unattended
device (CCD) camera to output a o if OIO is used, clean lenses with
digital image to a monitor xylene after use
 optical image is projected  How to Focus a Microscope
directly on CCD camera; entire o proper way to focus
system is designed for monitor  first, start with lowest power
image objective lens
 Use and Care of Microscope  while looking from side, crank
o transport from cabinet to working lens down as close to specimen
area as possible without touching it
 hold arm with one hand o look through eyepiece lens and
 other hand supports base focus upward only until image is
o place at least 6 inches away from sharp
edge of table  if you can’t get it in focus, repeat
o before using, clean first (with lens process again
paper / soft piece of cloth / gauze)  once image is sharp with low
 eyepiece power lens
 objectives - you should be able to simply
 condenser click in next power lens
 mirror - do minor adjustments with
o always see to it that mirror is focus knob
directed towards source of light o if it has a fine focus adjustment
o place LPO on center of stage by  turning it a bit should be all
moving revolving nosepiece until a that’s necessary
soft click is heard  continue with subsequent
o look into it and check is there is objective lenses and fine focus
enough light each time
o adjust mirror and diaphragm to  Magnification
secure proper amount of light o number tells operator how many
o place slide with specimen on stage times the focused specimen is
and always bear in mind to clip it magnified
o never tilt it when using a wet mount o computation – M = MPE x MPO
preparation or immersion oil  M – magnification
o focus using coarse adjustment knob  MPE – magnifying power of
 always focus upward or away eyepiece
from slide  MPO – magnifying power of
o shift to HPO or OIO, if necessary, objective
without elevating body tube
 focus using fine adjustment knob
only