RATIFICATION PAGE Complete report of Basic Biology with title “How to Use Microscope”, which made by: Name : Shally Rahmawaty Reg. Num (ID) : 1312440001 Class : Physics ICP A Group : II After checked by assistant and assistant coordinator, so this report accepted, Makassar, November 2013 Assistant Coordinator Assistant Adi Putra Rahman Nur Hafri Mulyani ID: 091414021 ID : 1114040204
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RATIFICATION PAGE
Complete report of Basic Biology with title “How to Use Microscope”,
which made by:
Name : Shally Rahmawaty
Reg. Num (ID) : 1312440001
Class : Physics ICP A
Group : II
After checked by assistant and assistant coordinator, so this report accepted,
Makassar, November 2013
Assistant Coordinator Assistant
Adi Putra Rahman Nur Hafri Mulyani
ID: 091414021 ID : 1114040204
Known,
Lecture of Responsibility
Sitti Zaenab, S.Pd, M.Pd
ID: 19810302 200912 2 003
CHAPTER IINTRODUCTION
A. Background
So many things that can not be seen with naked eye. Examples
constituent structure of living things. Bodies of living things (organisms) are
composed of cells. The cell is the smallest unit of an organism. Due to the very
small size, the cell can not be seen with the eyes directly so it takes a special tool
to be able to see it. This has led researchers to look for a tool that can be used to
see objects that are very small. Until then found that the microscope can be used
to see objects as small as a cell size.
Microscope is an instrument designed to make fine details visible. A
microscope is an object used to see objects that are too small to be viewed using
naked eyes. The purpose of a microscope is to enlarge and make the tiny objects
visible and it is mainly used in scientific research.
Historically, the study of cell biology could not have happened without the
invention of microscopes because cells were not known to exist before Antonin
van Leeuwenhoek and Robert Hooke saw them in their primitive microscopes.
The early simple “microscopes” which were really only magnifying
glasses had one power, usually about 6x – 10x. One thing that was very common
and interesting to look at was fleas and other tiny insects. These early magnifiers
were hence called “flea glasses”. The “simple microscope” or magnifying glass
reached its highest state of perfection, in the 1600’s, in the work of Anton von
Leeuwenhoek who was able to see single-celled animals (“animalcules”) and
even some larger bacteria. Meanwhile, the cells were first discovered by Robert
Hooke in 1655 by using a simple microscope. Robert Hooke saw cork cells the
cells of the incision.
The microscope consists of several components, namely optical and
mechanical components that have different functions. Before performing the
observations, we must know the function the parts of the microscope to help us in
making observations. In addition, we also need to know how to use, how to clean
and how to keep a microscope.
A. Purpose
The objective in this experiment is mean to skillfully use and safely
biological microscope to see the simple preparations.
B. Benefit
The benefits of this practicum is to determine, recognize microscope parts and
their functions, and how to use it. And understand the elements contained in the
object of research.
CHAPTER IIPREVIEW OF LITERATURE
The cell theory: Naturalists and scientists continued observing the living
microscopic world using glass lenses. In 1838, German scientist Matthias Schleiden
carefully studied plant tissues and concluded that all plants are composed of cells. A
year later, another German scientist, Theodor Schwann, reported that animal tissues
also consisted of individual cells. Prussian physician Rudolph Virchow proposed in
1855 that all cells are produced from the division of existing cells. The observations
and conclusions of these scientists and others are summarized as the cell theory. The
cell theory is one of the fundamental ideas of modern biology and includes the
following three principles:
1. All living organisms are composed of one or more cells.
2. Cells are the basic unit of structure and organization of all living
organisms.
3. Cells arise only from previously existing cells, with cells passing copies of
their genetic material on to their daughter cells. (Glencoe, 2008)
Cell walls were first seen by Robert Hooke in 1665 as he looked through a
microscope at dead cells from the bark of an oak tree. But it took the wonderfully
crafted lenses of Antoni van Leeuwenhoek to visualize living cells. Imagine Hooke’s
awe when he visited van Leeuwenhoek in 1674 and the world of microorganisms
what his host called very little animalcules was revealed to him. (Campbell, 2010)
A microscope is an instrument designed to make fine details visible. The
microscope must accomplish three tasks: produce a magnified image of the specimen
(magnification), separate the details in the image (resolution), and render the details
visible to the eye, camera, or other imaging device (contrast). Because of the limited
ability of the eye’s lens to change its shape, objects brought very close to the eye
cannot have their images brought to focus on the retina. The accepted minimal
conventional viewing distance is 10 inches or 250 millimeters (25 centimeters). More
than five hundred years ago, simple glass magnifiers were developed. These were
convex lenses (thicker in the center than the periphery). The specimen or object could
be focused by use of the magnifier placed between the object and the eye. These
“simple microscopes”, along with the cornea and eye lens, could spread the image on
the retina by magnification through increasing the visual angle on the retina. The
“simple microscope” or magnifying glass reached its highest state of perfection, in
the 1600’s, in the work of Anton von Leeuwenhoek who was able to see single-celled
animals (“animalcules”) and even some larger bacteria. The image produced by such
a magnifier, held close to the observer’s eye, appears as if it were on the same side of
the lens as the object itself. Such an image, seen as if it were ten inches from the eye,
is known as a virtual image and cannot be captured on film. These magnifiers had
severe limitations in specimen positioning, illumination, lens aberrations, and
construction. (Mortimer, 2003)
Three important parameters in microscopy are magnification, resolution, and
contrast. Magnification is the ratio of an object s image size to its real size. Light
microscopes can magnify effectively to about 1,000 times the actual size of the
specimen; at greater magnifications, additional details cannot be seen clearly.
Resolution is a measure of the clarity of the image; it is the minimum distance two
points can be separated and still be distinguished as two points. For example, what
appears to the unaided eye as one star in the sky may be resolved as twin stars with a
telescope, which has a higher resolving ability than the eye. Similarly, using standard
techniques, the light microscope cannot resolve detail ner than about 0.2 micrometer (
m), or 200 nanometers (nm), regardless of the magnification. The third parameter,
contrast, accentuates differences in parts of the sample. Improvements in light
microscopy have included new methods for enhancing contrast, such as staining or
labeling cell components to stand out visually. (Campbell, 2010)
Optical microscope consists of two, namely biological microscopes and stereo
microscopes. Biological microscope is used for observation of transparent thin
objects. Irradiation is given from below with natural light or lights. This biological
microscope eyepiece and generally has an objective lens with a magnification power
as follows :
1. 4x objective with 10x eyepiece, 40x magnification
2. Ocular 10x by objective 10x, 100x magnification
3. Ocular 40x by objective 10x, 400x magnification
4. 100x objective with a 10x eyepiece, magnification 1000x
Objective of the most powerful optical microscope objective emersi 1000x
called, because of its use must be with emersion oil and how to wear it especially
well. Stereo microscope is used for observation of objects that are not too large,
transparent or not. The rays can be set from above or from below with natural light or
lights. Has two objective and two ocular, in order to obtain a three-dimensional image
with the observation of two-piece eye. Strength is not too strong magnification 1x or
2x general objective with 10x eyepiece or 15 times. (Tim penyusun, 2013)
In a light microscope (LM), visible light is passed through the specimen and
then through glass lenses. The lenses refract (bend) the light in such a way that the
image of the specimen is magnified as it is projected into the eye or into a camera.
Until recently, the resolution barrier prevented cell biologists from using standard
light microscopy to study organelles, the membrane-enclosed structures within
eukaryotic cells. To see these structures in any detail required the development of a
new instrument. In the 1950s, the electron microscope was introduced to biology.
Rather than light, the electron microscope (EM) focuses a beam of electrons through
the specimen or onto its surface. Electron microscopes have revealed many organelles
and other subcellular structures that were impossible to resolve with the light
microscope. But the light microscope offers advantages, especially in studying living
cells. A disadvantage of electron microscopy is that the methods used to prepare the
specimen kill the cells. (Campbell, 2010)
Another type of microscope, the scanning tunneling electron microscope
(STM), involves bringing the charged tip of a probe extremely close to the specimen
so that the electrons “tunnel” through the small gap between the specimen and the tip.
This instrument has enabled scientists to create three-dimensional computer images
of objects as small as atoms. Unlike TEM and SEM, STM can be used with live
specimens. The atomic force microscope (AFM) measures various forces between the
tip of a probe and the cell surface. (Glencoe, 2008)
CHAPTER III
OBSERVATION METHOD
A. Time and Place
Day/date : Friday, November 22nd 2013
Time : 04.00-05.50 PM
Place : Green House of Biology, FMIPA UNM
B. Tools and Materials
1. Tools provided by laboratory
a. Biological microscopes
b. Toolbox contains of:
1) Glass object
2) Glass cover
3) The cup petri
4) Tweezers
5) Pipette
2. Tools provided by student
a. Razor blade
b. Flannel fabric
c. Roll tissue
d. Drawing book and pencil
e. Toothpick
3. Materials provided by laboratory
a. Distilled water
b. Filter paper or blotting paper
c. Cotton
4. Materials provided by student
a. Leaf of Allium cepa
b. Leaf of Hibiscus rosa-sinensis
c. Leaf of Sechium edule
C. Work Procedure
1. Setting up the microscope
1.1 The microscope is placed on the desk right in front of us.
1.2 Body of microscope cleaned with flannel. Never rub the lens with
flannel.
1.3 The tool box is opened, remove the cup containing the stained glass
objects and glass cover. Glass objects are cleaned with a cotton cloth or
filter paper.
1.4 On the table there are only a microscope work, toolbox with its contents,
the handbook and notes, materials for the lab. Other excluded at other
places that have been provided.
2. Regulate the entry of light into the Tubus
2.1 The state of lab space considered, where the direction of the brighter
light (from the front, left, or right), Mirror microscope light source is
directed to. Diaphragm plate is opened or rotated at a medium-size hole
position. Microscope condenser which has set its position to approach
the preparation and use flat mirrors. To microscope without condensers
used a concave mirror.
2.2 The position of the objective lens revolver arranged so that most short-
facing bench preparations until a click.
2.3 Tubus reduced to the distance to the objective end of the preparation
table or Tubus down 5-10 mm maximum.
2.4 We telescoped through the eyepiece with the left eye without squinting
(need practice) will appear white circular field.
2.5 Prepared microscope used to observe preparations.
3. How to adjust the distance of the lens with the preparation
3.1 With hands, coarse adjustment rotated or macrometer regulator towards
masters fingers, Tubus down, distance objective with a smaller
preparation table, do the opposite.
3.2 Glass objects containing preparations on the table amid hole desk
placed, glass objects clamped with clip so as not to shake.
3.3 Should be noted the distance objective with glass objects no more than
10 mm. If the distance was great, then rotated to lower micrometer
tubus while viewed from the side closer to the tip of the objective glass
objects up to a maximum 5-10 mm.
3.4 We telescoped through the eyepiece while hand turning macrometer by
raising tubus slowly.
3.5 Magnification of eyepiece and objective is checked. Calculated the
magnification of shadow.
3.6 Removed preparations that have been observed.
4. Making simple preparations
Observe the cotton fiber
4.1 The cleaned glass objects were taken and held evenly as possible.
4.2 Spilled glass of clear water or a drop of distilled water in the midst of.
4.3 With tweezers, one crate material removed and placed amid water
droplets.
4.4 The hand that helds the cover glass door between the master with the
index finger on the opposite side or edge.
4.5 Side of the cover glass on glass objects touched by water droplets with
a slope of 450 then released so right cover water droplets. Excess water
that seeps on the edge of the glass is absorbed by the filter paper.
4.6 Artificial preparations have been made, fitted on a table and observed
preparations as steps 3.2, 3.3, 3.4, and 3.5.
5. Observed enlargement
5.1 Shadow that appears to be raised again. Position preparations or tubus
not be touched.
5.2 Nosepiece rotated such that the objective lens longer (stronger)
perpendicular to the preparation table until there was a click.
5.3 We telescoped while turning the micrometer until the image appears
larger.
5.4 At the end of the activity using a microscope, there are some things
must be considered:
a. Mixture should not be stored on the table preparations, should be
excluded.
b. Mixture should be cleaned with a wet filter paper or cotton cloth
(glass objects + glass cover). Store in a petri dish and put in the gear
box.
c. Body of microscope is cleaned with a flannel cloth. Lowering tubus
low as possible.
d. Stored microscope into its box.
e. All the equipment has been used, cleaned with cotton cloth and kept
in a box.
f. Equipment itself, kept themselves to be used in the next activity.
g. The rest of the materials are not used anymore discarded rubbish