01 Basic Microscopy and Animal Cell Structure

BASIC MICROSCOPY AND ANIMAL CELL STRUCTURE
INTRODUCTION
Cells, as the basic unit of life, form the basic units of all living organisms. All animals are composed of
cells, a principle that was elucidated by Theodor Schwann in 1839. Most of the cells that compose
animal tissues and organs (with the exception of some egg cells) are small and invisible to the naked
eye. However, with the aid of a microscope, one can observe the basic animal cell structure. Cell
parts that can be observed using a compound light microscope include the cell membrane, nucleus,
and cytoplasm. Microscopy, therefore, is a very important laboratory technique in the study of animal
specimens.
OBJECTIVES
At the end of the laboratory activity, the students should be able to:
1. Determine the magnification of a microscope depending on the specific objective in place;
2. Estimate the size of a microscopic object;
3. Prepare a stained smear of animal cells; and
4. Identify an animal cell and their cell parts.
MATERIALS
Microscope
Transparent ruler
Ocular micrometer disc
Glass slide
Methylene blue
Dropper
Toothpick
Alcohol lamp
Matches/lighter
Prepared slide of a nervous tissue
METHODS
A. Magnification, Field of Vision, and Calibration Constant
1. Magnification: Calculate the total magnification of your microscope when using the a) low
power objective (LPO) and b) high power objective (HPO).
2. Diameter field of vision: Place a transparent millimeter (mm) ruler under the LPO. Estimate
the diameter of the field of vision in mm by counting the number of millimeter divisions you
see through the microscope. Record this measurement and convert to micrometer units (1
mm = 1000 m).
Calculate the field of vision with the high power objective using the following formula:
Diameter of field at high power = Magnification at low power x Diameter of field at low power
Magnification at high power
3. Calibration Constant. Replace your ocular with a micrometer eyepiece or insert a
micrometer disc inside the ocular. Place a ruler with millimeter divisions on the stage and
determine the number of micrometer spaces within one millimeter division when the LPO is
used. Make three measurements and get the mean number of micrometer spaces per
millimeter.
Compute for the calibration constant, C, for that micrometer eyepiece using the following
formula:
C = 1/X, where X = mean number of micrometer spaces per millimeter
B. Preparation of a Stained Smear
1. Put a drop of methylene blue on a clean glass slide.
2. Obtain squamous cells from the walls of your oral cavity using the blunt tip of an unused
toothpick. Mix the tip of the toothpick into the drop of methylene blue on the glass slide.
3. Pass the slide through the flame until the drop dries completely.
4. Observe squamous cells under LPO and HPO. Draw your observation. Label the following
structures: cell membrane, nucleus, and cytoplasm.
C. Estimating the Size of a Microscopic Object
1. Place the stained smear of squamous cells under the microscope.
2. Estimate the size of a cell by counting the number of micrometer spaces that span its entire
diameter. Multiply this by the calibration constant. This will give the size in millimeters.
Convert this to micrometers.
3. Repeat the measurements thrice and get the average.
D. Observation of Animal Cells
1. View a prepared slide of nervous tissue under the microscope. Observe the slides under both
LPO and HPO. Draw and label the following representative parts of the neuron as seen under
the microscope: cell membrane, cytoplasm, nucleus, dendrites, and axon.
RESULTS
A. Magnification and Field of Vision
Table 1. Magnification of Microscope No. _____
Objective Lens
Ocular Lens
Magnification
Scanning power objective
Low power objective (LPO)
High power objective (HPO)
Objective Lens
Magnification
Table 2. Diameter Field of Vision of Microscope No. _____

Objective
Diameter Field of Vision
Lens
(in m)
Low power objective (LPO)
High power objective (HPO)
Table 3. Calibration Constant of Microscope No. _____
Trial Number Number of Micrometer Spaces per mm
1
2
3
Mean number of micrometer spaces per mm:
Calibration constant, C:
Total
Magnification
B. Preparation of Stained Smear
Specimen: _______________
Magnification: x ___________
Specimen: _______________
C. Estimating the Size of a Microscopic Object

Table 4. Size of a Squamous Cell
Trial Number Calibration
Constant
1
2
3
Number of Micrometer
Spaces
Size
(in mm)
Size
(in m)
Specimen: _______________
Average size (in m): _______
D. Observation of Animal Cells
Specimen: _______________
Specimen: _______________
QUESTIONS
1. How would an eyepiece of 5X change total magnification in relation with the two objectives
lenses (i.e., LPO and HPO)?
2. Which objective lens covers a wider field, LPO or HPO? Explain.
3. What cells parts did you observe in the stained smear of squamous cells? In the prepared
slide of nervous tissue? Describe these parts as seen under the microscope.
REFERENCES
Angtuaco SP, Carumbana EE, Pena-Muralla RM, Yago RE. 1994. Zoology: A Laboratory Manual.
Quezon City: Ateneo de Manila University Office of Research and Publications. 109 p.
Hickman CP, Roberts LS, Keen SL, Eisenhour DJ, Larson A, Anson H. 2011. Integrated Principles of
th
Zoology. 15 ed. New York (NY): McGraw-Hill. 842 p.

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BASIC MICROSCOPY AND ANIMAL CELL STRUCTURE

Table 2. Diameter Field of Vision of Microscope No. _____

B. Preparation of Stained Smear

C. Estimating the Size of a Microscopic Object

2. Which objective lens covers a wider field, LPO or HPO? Explain.

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