EXPERIMENT 1 : MICROSCOPE – EXPLORING AND VIEWING

OBJECTIVE :
1) To identify the parts of compound and stereoscopic microscopes and be proficient in their
correct use in biological studies.
2) To learn the technique to measure the size of a specimen using a stage micrometer and an
ocular micrometer.

ABSTRACT :
This lab experiment shows the students that it is very important to learn the tools and functions that
will be used to perform an experiment. In this lab, students will be able to know the differences
between the compound microscope and stereoscopic microscope. Furthermore, students will also be
able to learn how to calibrate the measurement to ensure correct measurement and calculation of the
sample with a microscope. They will also measure the size of a specimen using a micrometer and an
ocular micrometer such as leaf, pond water, insects, and prepared slides. On this last task, students
will examine all the prepared slides under the best magnification, drawing a cell at the different
magnificatioms levels and measuring cells.

INTRODUCTION :
A microscope is an instrument used to see objects that are too small to be seen by the naked eye.
Microscopy is the science od investigating small objects and structures using the instrument. The
stereoscopic is an optical microscope for low magnification observation of a sample, typically using
light reflected from the surface of an object rather than transmitted through it. Next,compound
microscope uses a lens close to the object being viewed to collect light which focuses a real image of
the object inside the microscope. That image is then magnified by a second lens or group of lenses
that give the viewer an enlarged inverted virtual image of the object. The objectives for the
experiments to acquire the skill of calibrating prior to measure field of view and cell size using the
ocular and stage micrometer. The prepared slides can be viewed with a microscope to produce an
enlarged image and others specimens such as leaf, pond water and insect can be viewed with a
dissecting microscope.

MATERIALS
-Compound microscope
-Stereoscopic microscope

-Specimens (leaf, pond water, insects, prepared slides)

- Distilled water

-Cover slips

-Tissue paper

-Glass slides

-Forceps

-Razor blades
-Ocular and stage micrometer

-Dropper

-Petri-dishes

-Oil immersion

Task 1 : Identify the parts of compound an stereoscopic microscopes

Method
1. Carefully obtain the stereoscopic and compound microscope from the display cabinets. One
person from each group will go over to the microscope storage area and properly transport
one microscope to our working area. Be sure to handles microscope with care as they are
delicate, expensive tools and must never experience rough handling or be dropped. Carry the
microscope with both hands which are one hand on the arm and the other under the base of
the microscope.
2. One person from our group will pass over one of the glass slide.
3. Turn on the microscope and place the slides on the stage and fasten it with stage clips. You
can push down the back end of the stage clip to open it.
4. The nosepiece slide is rotated to the lowest-power objective. For an example, 4x objective
lens.
5. The coarse adjustment knob is adjusted to the largest diameter, allowing the greatest amount
of light to pass through.

6. Slowly turn the coarse adjustment so that the objective lens goes up (away from the slide).
Continue until the image comes into focus. Use the fine adjustment, if available, for fine
focusing. If you have a microscope with a moving stage, then turn the coarse knob so the
stage moves downward or away from the objective lens.
7. Move the microscope slide around so that the image is in the centre of the field of view.
8. Now, you should be able to change to the next objective lenses with the only minimal use of
the focusing adjustment. Use the fine adjustment, if available. If you cannot focus on your
specimen, repeat steps 4 through 7 with the higher power objective lens in place. (Precaution:
Do not allow the objective lens to touch the slide)
9. When finished, raise the tube (or lower the stage), click the low power lens into position and
remove the slide.
RESULT

STEREOSCOPIC MICROSCOPE

1 2
8
5
4

9
6
7

COMPOUND MICROSCOPE
1

5
4
6
8
7
9
23
10
TASK 2 : MEASURING THE SIZE OF A SPECIMEN USING A STAGE
MICROMETER AND AN OCULAR MICROMETER AND IMMERSION
OIL MICROSCOPY (SPINAL CORD )
METHOD
1. The stage micrometer is placed on the stage of the microscope.
2. After the focus on the stage micrometer had been done, the eyepiece is rotated until the ocular
micrometer is aligned to the stage micrometer.
3. Using the 10x objective lens, the ocular lens is adjusted so that it is aligned directly to the stage
micrometer. At 25 ocular divisions, it occupied 0.1 mm on the stage micrometer.
4. Using the information obtained, the calibration factor for 10x objective lens had been acquired.
5. Repeat Step 3 and 4 using objective lens of 4x, 10x, 40x and 100x.
6. After obtaining calibration factor for each objective lenses, the estimated size of the sample or cell
by using the information of the length of the sample or cell in O.D. and use it in the calibration factor,
depending on the objective lens used.
7. For immersion oil microscopy, the turret was rotated to 40x objective, located the desired portion of
the specimen spinal cord in the center of the field. The specimen was refocused very carefully as
sharply as possible. ( Do not alter focus for the following steps )
8. The critical step- Turret was rotated partially so that 40x and 100x objectives straddle the specimen.
A small dropped of oil was placed on the slide in the center of the lighted area.
9. The turret was rotated 100x oil immersion objective touches the oil and clicked into place. Focus
only with fine focus. The specimen will come into focus easily. With more than one specimen on a
slide, the focusing was not altered, a drop of oil was placed on the second specimen and laterally slide
the slide until it is in place. (Never go back to the 10x 40x objectives after we have applied oil to the
specimen since oil can ruin the lower power objectives.
RESULT

i) Calibration Factor

 40x Total Magnification

40ocd=1000mm
1ocd =25 mm

 100x Total Magnification

1ocd = 10 mm

 400x Total Magnification

1 ocd = 2.5 mm

 1000x Total Magnification

1 ocd = 1.0 mm
INSECT ( ANT )
10x Magnification lens
Cell size : Antenna 300 mm equal to 3ocd

LEAF
40x Magnification
Cell size : 275 mm equal to 11 ocd
POND WATER
10x magnification
Cell size : 2000mm equal to 20 ocd
We can find fibre in the cell.

SPINAL CORD
4x Magnification
Size : 7500 mm equal to 30 ocd
DISCUSSION
The compound microscope is the most common type of microscope, contains several parts
with the specific function. Each part of the microscope has their own role in order to make
microscope function as explained in the result. Total magnification of an image can be
calculated by multiplying the magnification of the objective lens by the magnification of the
ocular lens. For example, the magnification of the ocular lens is usually 10x, while the
magnification of objective lens is 40x. To get the total magnification is, 10x multiply by 40x
which the answer is 400x. So the total magnification is 400x. The standard objective lenses
fixed on the turret of the microscope are 4x, 10x, 40x and 100x objectives. The 4x, 10x and
40x lenses are collectively known as the dry lens. The exception about 100x objective is
known as the oil immersion as it requires the use of immersion oil because you cannot use
immersion lens without oil. Never get oil on any other lenses. When you already use the 100x
objective, you cannot turn back to use 4x, 10x, 100x. In order to measure the size of
magnified object, one must use an ocular micrometer which is located at one of the lenses of
the microscope.Since the conversiom factor different for each magnification, the calibration
factor of different objective lense must be calculated individually, the calibration of the
microscope is measured when the ocular and stage microscope are aligned together. In our
microscope the scale of ocular microscope is 1 to 5ocd while the stage micrometer consists
0.01mm in each space. The calibration of the microscope is 40ocd equal to 1000micrometer.
Therefore, 1ocd is equal to 25micrometer. The calibration that stated above is a scale to
measure the specimens. For example, the observation on the specimen of pond water contains
a fiber is measured at the tip of the fiber at 10× magnification give a result 20ocd.Then the
20ocd is multiplied with 100micrometer because 1ocd is equal to the 100micrometer. Thus,
the result is 2000micrometer. The technique is repeated for all specimens based on their
magnification.

CONCLUSION
In conclusion, the microscope can help us to observe the thing, which is not visible to the
naked eye, allows us to see objects magnified. There are many other types of microscope
other than compound microscope and dissecting microscope. All of the parts have their own
functions. By calibrating the ocular micrometer with the stage micrometer, one could
somewhat accurately sum up or estimate the cell size of the given sample.

REFERENCES
Groshon, N. (2017). Calibrating Your Microscope. Retrieved October 2, 2018 from
https://www.mccrone.com/mm/calibrating-microscope/
Zamboni, J. (2018). What Is the Function of a Microscope. Retrieved October 1, 2018 from
https://sciencing.com/function-microscope-6575328.html
The complete Microscope Guide. (2013). Retrieved September 29 2018, from Best
Microscope Reviews, Information & Microscopy Research:
http://www.microscopemaster.com/
 FACULTY OF APPLIED SCIENCES

EXPERIMENT 1 : MICROSCOPE – EXPLORING AND VIEWING

NAME : AQILAH BINTI AZMAN

STUDENT ID: 2018440126

GROUP : AS2461B
DATE OF EXPERIMENT : 28 SEP 2018
DATE OF SUBMISSION : 5 OCT 2018
LECTURER’S NAME : PROF MADYA DR MANSUR AHMAD
OBJECTIVE:
1. Identify the reducing sugar using Benedict’s reagent
2. Identify the presence of starch using IKI stain

ABSTRACT:

The experiment is designed to test macromolecules. One example for macromolecules is

carbohydrate. In order to do the test, specific reagents are used. The result will show specific
colour changes based on the macromolecules and the types of macromolecules can be
verified.

For experiment of simple sugar, Benedict’s solution is used as the reagent. A prediction is
made that glucose and maltose are reducing sugar and will change the blue colour of
Benedict’s reagent to brick red precipitate and orange colour respectively.

For experiment to test sugar in Pepsi drink, IKI stain and Benedict’s reagent are used. When
pepsi is mixed with IKI stain, it is predicted to change brown colour solution of pepsi to
orange in colour, and cloudy orange after heating.

For experiment of starch, iodine is used as the reagent. The yellow colour of iodine is
predicted to change to dark blue when put on potato strip.

INTRODUCTION

Biomolecules are molecules that occur naturally in living organisms. The basic types
of molecules are carbohydrates, lipids, proteins and nucleic acids. The chemistry of these
carbon-based molecules is included in the field of organic chemistry with the specific life-
related processes forming the field of biochemistry. Biomolecules are very large molecules
consists of many atoms, that are bound together.
All of these biomolecules are used to run our metabolic processes. Carbohydrates
include both sugar and polymers provide the body with source of fuel and energy, it aids in
proper functioning of our brain, nervous system, digestive and immune system. Deficiency of
carbohydrates in the diet causes fatigue, poor mental function. Carbohydrates serve as energy
sources and provide structural support as in the cell wall or plants. Carbon, hydrogen and
oxygen are the elements found in carbohydrates. Benedict’s reagent is used as a simple test
for determine reducing sugar. A reducing sugar is a carbohydrate possessing either a free
aldehyde or free ketone functional group as part of its molecular structure. Functional groups
are the regions of a molecule that gives it particular properties. A single molecule can have
more than one functional group.
DATA

Table 1: Benedict’s test for reducing sugar

Test tube Contents Observations
1 Water Blue → Blue
2 Glucose Blue → Brick red precipitate
3 Sucrose Blue → Blue
4 Maltose Blue → Orange
5 Starch Blue → Blue

Table 2: Test for sugar in Pepsi drink

Substance Before heating Observations After heating
Pepsi + IKI Brown colour solution Colour slightly Turn to orange
stain change from brown cloudy
to orange
Pepsi + Brown colour solution No change -
Benedict’s
reagent

Table 3: IKI test for starch

Substances Before After
Observations
Potato strip Yellow colour Dark blue colour

.
BEFORE HEATING

glucose maltose
sucrose
water
starch
 glucose
water

CONCLUSION:
In conclusion, reducing sugars are identified by using Benedict’s reagent, when
blue colour of Benedict’s reagent changes into brick red precipitate and orange.
The presence of starch is also identified in the potato strip by using IKI stain as
change the yellow colour of iodine into dark blue colour.

REFERENCES:
waterUrry,
1) Campbell, Reece, glucose
Cainsucrose
Wasserman, Minorsky,
starch Jackson.
maltose(2017).
Biology A Global Approach. Pearson
2) http://en.m.wikipedia.org/wiki/Biomolecule