THE COMPUND MICROSCOPE

By: Jhean Alphonsine T. Means

The Compound Microscope…

- Is an optical instrument having more than one lens that is used to enlarge small
objects.
- Has a resolution of 0.2µm (1,000x better than the human eye)?
- It consists of a two-lens system:
Objective Lens
Ocular/Eyepiece Lens

Terms used in Microscopy:

1. Parfocal: Having multiple lenses that have corresponding focal points in the same
plane
2. Working distance: is the distance between the objective lens and the specimen. At
low magnification, the working distance is relatively long. As you increase the
magnification the working distance decreases dramatically.
3. Field of View: Sometimes abbreviated "FOV", it is the diameter of the circle of light
that you see when looking into a microscope. As the power gets greater, the field of
view gets smaller
4. Micrometer: ls called a micron it is the metric linear measurement used in microscopy.
There are 1000 microns in a millimeter. If something is 1.8mm long then it can also
be expressed as 1,800 microns (or micrometers) long.
*How can eye strain be avoided in microscopy?
- maintain distance between your eye and eye piece of the microscope
- taking breaks
- it is important not to turn the microscope mirror towards the sun or other
sources of intense light, as the mirror amplifies the light.
*When is the concave surface of the mirror used?
- Concave mirrors are placed at the bottom of microscopes to catch light which
can be turned in any direction. Concave mirrors reflect light inward to one focal
point. They are used to focus light.
- It's purpose to reflect light from a light source and amplify the light.
Features of the Compound Microscope:
• Magnification – is the number of times an object has been enlarged
* The total magnification is derived by multiplying the magnifying power of the objective
lens and the magnifying power of the ocular lens. *
• Revolving Power/Resolution – refers to the ability of the lenses to distinguish two
separate objects at a specified distance apart as two distinct objects
Components of the Compound Microscope:
• The three basic, structural components of a compound microscope are the head, base
and arm.
Head/Body: houses the optical parts in the upper part of the microscope
Base: (of the microscope) supports the microscope and houses the
illuminator
Arm: connects to the base and supports the microscope head. It is also
used to carry the microscope.
Optical Components of the Compound Microscope:
• There are two optical systems in a compound microscope: Eyepiece Lenses and Objective
Lenses:
Eyepiece or Ocular: is what you look through at the top of the microscope.
Typically, standard eyepieces have a magnifying power of 10x. Optional eyepieces
of varying powers are available, typically from 5x-30x.
Objective Lenses are the primary optical lenses on a microscope. They range from 4x-
100x and typically, include, three, four or five on lens on most microscopes. Objectives
can be forward or rear-facing. One of the most important parts of a compound
microscope, as they are the lenses closest to the specimen.
Eyepiece Tube: holds the eyepieces in place above the objective lens. Binocular
microscope heads typically incorporate a diopter adjustment ring that allows for the
possible inconsistencies of our eyesight in one or both eyes. The monocular (single eye
usage) microscope does not need a diopter. Binocular microscopes also swivel
(Interpupillary Adjustment) to allow for different distances between the eyes of different
individuals.
Diopter Adjustment: Useful as a means to change focus on one eyepiece so as to correct
for any difference in vision between your two eyes.
Coarse adjustment: Brings the specimen into general focus.
 Fine adjustment: Fine tunes the focus and increases the detail of the specimen.
Nosepiece: A rotating turret that houses the objective lenses. The viewer spins
the nosepiece to select different objective lenses.
Specimen or slide: The specimen is the object being examined. Most
specimens are mounted on slides, flat rectangles of thin glass.
The specimen is placed on the glass and a cover slip is placed over the
specimen. This allows the slide to be easily inserted or removed from
the microscope. It also allows the specimen to be labeled, transported,
and stored without damage.
Stage: The flat platform where the slide is placed.
Stage clips: Metal clips that hold the slide in place.
Stage height adjustment (Stage Control): These knobs move the stage left
and right or up and down.
Aperture: The hole in the middle of the stage that allows light from the
illuminator to reach the specimen.
On/off switch: This switch on the base of the microscope turns the
illuminator off and on.
Illumination: The light source for a microscope. Older microscopes used
mirrors to reflect light from an external source up through the
bottom of the stage; however, most microscopes now use a
low-voltage bulb.
Iris diaphragm: Adjusts the amount of light that reaches the specimen.
Condenser: Gathers and focuses light from the illuminator onto the specimen
being viewed.

How does a Microscope work?

• All of the parts of a microscope work together.
• The light from the illuminator passes through the aperture, through the slide, and through
the objective lens, where the image of the specimen is magnified.
• The then magnified image continues up through the body tube of the microscope to the
eyepiece, which further magnifies the image the viewer then sees.
 • Mechanical Parts – used to support and adjust the parts
Base
Body Tube
Arm
Revolving Nosepiece
Stage and Slide Holder
Diaphragm lever
Course Adjustment knob
Fine Adjustment knob

 Illuminating Parts – used to provide lights

Mirror
Electric Lamp
Iris Diaphragm
Condenser

 Magnifying Parts – metal cylinders attached below the nosepieces and contains especially
ground and polished lenses
Low Power Objective
- magnifies about 10x
- shortest lens
- provides 4-fold magnification and is frequently called the 4X
High Power Objective
- magnifies about 40x
- achieves higher levels of magnification
- used to view smaller specimens such as cell structures which cannot be seen at
lower levels of magnification
OIO / Oil Immersion Objective
- give the highest magnification, usually 97% or a 100%.
- used wet either with cedar wood oil or synthetic oil
- are used only at very large magnifications that require high resolving power.
- Objectives with high power magnification have short focal lengths,
facilitating the use of oil. The oil is applied to the specimen (conventional
microscope), and the stage is raised, immersing the objective in oil.

When is the oil immersion objective used and what kind of oil is used?
• a technique used to increase the resolving power of a microscope
• achieved by immersing both the objective lens and the specimen in a transparent oil of
high refractive index.
 • Immersion oils are transparent oils that have specific optical and viscosity characteristics
necessary for use in microscopy.
OTHER TYPES OF MICROSCOPES:
• Optical microscope: A microscope that allows easy observation of 3D objects at low
magnification.
• Electron microscope: These microscopes emit electron beams, not light beams, toward
targets to magnify them.
• Scanning probe microscope (SPM): This microscope scans the surface of samples with a
probe and this interaction is used to measure fine surface shapes or properties.
• Others: X-ray microscope, ultrasonic microscope, etc.

DIFFERENT TYPES OF MICROORGANISMS SEEN UNDER HPO:

BACTERIA
• Bacillus subtilis - is also known as hay bacillus or grass bacillus. It is a gram-positive
bacterium and commonly found in soil and the GI tract of ruminants and humans. Bacillus
subtilis is rod-shaped and typically 4-10 microns long.
- Discovered by Christian Gottfried Ehrenberg
- is only known to cause disease in severely immuno-compromised patients and can
conversely be used as a probiotic in healthy individuals. The bacteria rarely cause food
poisoning.
- used to produce lipases, amylase and proteases, enzymes used in breakdown of complex
molecules and fats in waste water treatment and remediation
- bacterium is resistant to extreme temperatures, it can with stand high cooking
temperatures.
- It is predicted that it spends most of it time inactive and in spore form. When the
bacterium is active though, it produces many enzymes. One enzyme contributes to the
plant degradation process.
- Used in treatment of human diseases and to stimulate IgM, IgG and IgA antibodies in
immune suppressed cancer patients
- Bacillus subtilis also produces a toxin called subtilisin. Subtilisin can cause allergic
reactions if there is repeated exposure in high concentrations.
 - commercial applications of Bacillus subtilis include cleaning agents in detergents, in de-
haring and batting in the leather industry, in the production of special Japanese and
Korean food, starch modification, the de-sizing of textiles, and other specialized
chemicals.
- Bacilli cause an array of infections from ear infections to meningitis, and urinary tract
infections to septicemia. Mostly they occur as secondary infections in immunodeficient
hosts or otherwise compromised hosts. They may exacerbate previous infection by
producing tissue-damaging toxins or metabolites that interfere with treatment.
• Spirillum: is a genus of Gram-negative bacteria within family Spirillaceae. are thought to
have been first described by van Leeuwenhoek in the 1670s and later by Muller.
- large chemotrophic spirilla 1.4 to 1.7 micrometers in diameter and up to 60 micrometers
in length. They have rigid helical cell structure as opposed to the flexible cell structure of
spirochetes.
- have large, elongated rigid spiral structure, wavy
- cause bacterial diarrhea, adenocarcinoma and peptic ulcers.
- normal flora of the pharynx, mouth and lower GI
- There are two species in Spirillum with standing in nomenclature,
Spirillum winogradskyi: is a gram-negative, bacterium from the genus of Spirillum
which was isolated from the sulfidic sludge of a municipal wastewater treatment
plant.
Spirillum volutans: is a gram-negative, bacterium from the genus of Spirillum
which occurs in freshwater. Spirillum volutans is one of the largest bacteria
species.
• Staphylococcus aureus
- Microscopic appearance:
Cocci in grape-like clusters by irregular cell divisions in three plains
- Clinical significance:
Frequently found as part of the normal skin flora on the skin and nasal passages
and pharynx
It is estimated that 20% of the human population are long-term carriers of it.

- Staphylococcus is one of the five most common causes of infections after injury
or surgery.
- discovered by Sir Alexander Ogston in pus from surgical abscesses
- S. aureus is transmitted through air droplets or aerosol.
 - Of the variety of manifestations S. aureus may cause:
Minor skin infections, such as pimples, impetigo etc.
It may cause boils (furuncles), cellulitis folliculitis, carbuncles
It is the cause of scalded skin syndrome and abscesses
It may lead to lung infections or pneumonia
Brain infections or meningitis
Bone infections or osteomyelitis
Heart infections or endocarditis
Generalized life threatening blood infections or Toxic shock syndrome (TSS),
bacteremia and septicaemia
• Streptococcus : is a genus of coccus (spherical) Gram-positive bacteria that - groups in
chains that resemble a string of beads.
- are members of the normal flora (throat) and group of spheroidal
bacteria; are nonmotile
- They can spread through airborne droplets when someone with the infection coughs or
sneezes, or through shared food or drinks
Streptococci are divided into two key groups:
- Group A strep: are often found on the surface of the skin and inside the throat. They
are a common cause of infection in adults and children.

They can be spread in droplets in the coughs or sneezes of someone with an

infection, or through direct contact with an infected person or contaminated
object.
- Group B strep: usually live harmlessly inside the digestive system and in the
vagina.
can sometimes cause urinary tract infections (UTIs), skin infections, bone
infections, blood infections and pneumonia, particularly in vulnerable people,
such as the elderly and those with diabetes.
A strep B infection during pregnancy can also cause miscarriage or stillbirth, but
this is rare.
PROTOZOA
• Amoeba proteus: appears as an irregular, colorless, translucent mass of living animal-like
jelly or protoplasm that is constantly changing its shape by sending out and withdrawing
finger-like processes, the pseudopodia / “FALSE FEET”. Previously called Chaos diffluens.
- Discovered by Johann Rösel von Rosenhof, who named his discovery "Der Kleine
Proteus" ("the Little Proteus")
- The average size of an Amoeba proteus varies around from 0.2 to 0.3 mm in
diameter but larger forms have been found measuring up to .5 mm in diameter
which is visible to the eye.
-it possesses no cell wall, it has a thin delicate outer membrane called the plasma lemma.
- Amoeba proteus prefers to habitat clean ponds of highly oxygenated fresh water. It
is found in large food webbed ecosystems that contain lots of algae and plants. Since it is
adverse to light it will take cover under anything that provides shade.
- Its ability to change shape is from the pseudopodia, which are common in eukaryotic
microbes. The pseudopodia grants the microbe an ability to extend and contract into any
possible shape.
- The Amoeba proteus exists in fresh-water aquatic environments (ponds, streams,
puddles), or moist environments dominated by soil or plants.
- The Amoeba proteus is non-pathogenic, and is not capable of causing disease in other
organisms. But can serve as a host for a pathogenic infection.
- Of six species found in the human alimentary tract, Entamoeba histolytica causes amebic
dysentery, amoebic meningoencephalitis, and amoebic liver abscess.
Amoebic dysentery is caused by eating food or drinking water that has been
contaminated by amoebic cysts found in raw sewage. Also called as amoebiasis.
Amoebic meningoencephalitis is a rare but terrible disease caused by the
Naegleria fowleri amoeba. In this disease, the amoebas attack the central nervous
system.
Amoebic liver abscess: is the most frequent extraintestinal manifestation of
Entamoeba histolytica infection. an important cause of space-occupying lesions of
the live
• Paramecium: is a genus of unicellular ciliates. Paramecia are widespread in freshwater,
brackish, and marine environments and are often very abundant in stagnant basins and
ponds. Are not known to cause any diseases or infections.
 - known to Antonie van Leeuwenhoek, and were clearly described by his contemporary
Christiaan Huygens.
- "Animalcules which have no visible limbs or tails, and are of an irregularly oblong figure.“
- John Hill
- "Slipper animalcule," illustrated by Louis Joblot which he discovered in an infusion of oak
bark in water.
- Species of Paramecium range in size from 50 to 330 micrometres (0.0020 to 0.0130 in)
in length
- Cells are typically ovoid, elongate, foot- or cigar-shaped.
- The body of the cell is enclosed by a stiff but elastic membrane (pellicle), uniformly
covered with simple cilia, hairlike organelles which act like tiny oars to move the organism
in one direction.
- Paramecia live mainly by heterotrophy, feeding on bacteria, algae and yeasts. (They are
not capable of producing their own food. Therefore, they obtain their energy
requirements by feeding on organic matter or another organism.)
- Paramecium reproduces asexually, by binary fission.
ALGAE
• Spirogyra: is a genus of filamentous charophyte green algae, named for the helical or
spiral arrangement of the chloroplasts
- Belongs in the plantae kingdom classification.
- common names include water silk, mermaid's tresses, and blanket weed and there are
more than 400 species of Spirogyra in the world.
- measures approximately 10 to 100 μm in width and may grow to several centimeters in
length.
- very common in relatively clean eutrophic water, developing slimy filamentous green masses
- Spirogyra can reproduce both sexually and asexually
- made of green algae that have spiral chloroplast arrangements and
- primarily live in freshwater environments
- not known for causing diseases.
• Volvox is a genus of chlorophyte green algae in the family Volvocaceae. It forms spherical
colonies of up to 50,000 cells. They live in a variety of freshwater habitats, and were first
reported by Antonie van Leeuwenhoek.
- Each little alga within the colony bears two flagella, whip-like hairs and have anterior
eyespots that enable the colony to swim towards light.
- Volvox is facultatively sexual and can reproduce both sexually and asexually
- Two common species :
Volvox aureus
- is usually smaller and has less individual cells.
- colonies can also be visible with a size of half a millimeter as small green
dots.
- cells of Volvox aureus are also a bit more egg-shaped.
- the individual cells size are 5 to 8 micron
Volvox globator
- may reach a size of 2 millimeters so they can be easily seen with the naked eye.
If you are short sighted the larger Volvox aureus
- Individual cells are slightly smaller (4 micron
- One of the first things that you notice on Volvox is that most colonies have spheres
inside. These are 'daughter' colonies, called gonads which is a means of asexual
reproduction.
- Volvox also has a sexual reproduction. As in the asexual reproduction special cells are
formed around the equator. These cells develop into special germ cells.
- Male and female colonies form different germ cells. Sperm cells are formed by division.
The female germ cells do not divide. They enlarge to form ova.
- Volvox is a deadly pathogen of humans and related primates. The Volvox life cycle
involves entering a primate through a mucus membrane, from whence it enters the blood
stream, eventually making its way to the heart, brain, liver, or kidneys.
- Once lodged in an organ, Volvox begins converting mammalian tissue into juvenile
Volvoxes through a process called autocolony formation. The affected organ is slowly and
agonizingly devoured from within. Known symptoms include vomiting, neurosis, kurtosis,
sclerosis, gnosis, death and voting Republican.