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Activity 1

Review of Microscopy
Group 3
Castasus, Nathaniel
Dela Cruz, Jonabelle
Del Rosario, Louise
Diosomito, Allen
Prepared slides
Introduction
Microscope




MICROLAB

Philippines


MICROLAB

Philippines

History



Parts &
Functions



MICROLAB

Philippines

Types of
Microscope


Importance



Objectives
& Materials


Procedures



Worksheet




MICROLAB

Philippines


MICROLAB

Philippines


MICROLAB

Philippines


MICROLAB

Philippines


MICROLAB

Philippines
Microscope
mikrs "small" and skopen, "to look" or "see"
is an instrument used to see objects that are too
small for the naked eye.
Microscopy
The science of investigating small objects using such
instrument. Microscopy is the technical field of using microscopes
to view samples and objects that cannot be seen with the unaided
eye.
History of the Microscope
Roman's made the objects appear
larger through glasses

Salvino D'Armate made the
first eye glass,
The earliest simple forms
of magnification were
magnifying glasses,
usually about 6x - 10x
Who invented the microscope?

With the advancement of technology and improved
optics, the microscope as we know it today came into
being.
270x
(9x)
Parts & Functions
What makes microscopy a crucial
importance in the field of
microbiology?
Access to the existence of microorganisms

To observe, to manipulate, and to examine diversity of
microorganism through dimensions

Comparison between the different representative microorganisms
in shapes and sizes








- SIMPLE MICROSCOPE
- LIGHT COMPOUND MICROSCOPE
- ELECTRON MICROSCOPE
- SCANNING ELECTRON MICROSCOPE
COMMONLY USED
MICROSCOPES
OBJECTIVE S
REVIEW THE CONCEPT OF MICROSCOPY
CALCULATE THE MEASUREMENT OF
MICROORGANISMS
COMPARE THE SIZES AND SHAPES
MICROBIAL DIVERSITY
MATERIALS
COMPOUND MICROSCOPES
MONOCULAR
BINOCULAR
OCULAR MICROMETER
STAGE MICROMETER
PREPARED SLIDES
Aspergillus
Amoeba
Euglena
Bacillus

IMMERSION OIL
LENS PAPER
XYLENE
COTTON
PROCEDURES
MAGNIFICATION

Microscope was
plugged in
Light was switched
ON
The prepared slide
(Amoeba) was
placed on stage
The specimen area
of the slide was
placed over the
center of the stage
aperture
Coarse knob
adjustment was
raised until image
appears
Image was focused,
iris diaphragm
reduced for best
contrast.
Image was observed
in High Power
Objective. Image
sharpened.
Eyes kept at certain
distance from
eyepiece.
Amoeba proteus
400x
RESOLUTION
Bacillus subtilis slide
viewed under Low
Power Objective
Bacteria was focused
under High Power
Objective
Oil Immersion Objective
was shifted in place.
Focused using fine
knob
Small drop of
immersion oil was
applied on the
center of stage
aperture.
Front lens was
immersed in oil and
touched the slide.
Image was focused
using fine adjustment
knob.
After using OIO, the oil
was blotted with lens
paper with xylol.
Measurement of Specimen
A microscope with
ocular micrometer was
used.
The grid lines were
observed in upright
position using LPO.
The stage micrometer
was placed on the
stage and focused on
scale
The first division
coincided with the
division on stage scale.
Number of divisions were
counted on the ocular
micrometer subtended by
number of divisions on
stage micrometer.
Calibration factor for
the ocular unit was
computed.
Answers to Worksheet


Specimen no. 1
Amoeba proteus


Total Linear Magnification:
100x (LPO)
1. Draw the specimen under (a) LPO and (b) HPO
Worksheet
Specimen no.1
Amoeba proteus

Total Linear Magnification:
400x (HPO)
2. How does a microscope magnifies the
image of an object?

3. What characteristic of a glass lens is
responsible for its magnification?

SPECIMEN NO.2
Bacillus subtilis
Total linear magnification:
1000x (OIO without oil)
4. Draw the specimen (a) without Oil Immersion and (b) with Oil Immersion
Specimen no.2
Bacillus subtilis
Total Linear Magnification:
1000x (OIO with oil)
5. Why is it necessary to add immersion oil
on the slide being focused when using
OIO?
Objectives Number of OM
division subtended
by SM division
Number of SM
division subtended
by OM division
Value of 1 OM
division (in
microns)
LPO 20 100 50
6. Compute for the value 0f 1 OM division and fill in Table 1.1.
X(OU) = Y(SU)
6. Compute for the value 0f 1 OM division and fill in Table 1.1.
Objectives Number of OM
division subtended
by SM division
Number of SM
division subtended
by OM division
Value of 1 OM
division (in
microns)
Scanner 8 100 125
LPO 20 100 50
HPO 20 25 12.5
OIO 20 10 5
Total Magnification: 100x (LPO)
Objectives Size of 1 box
Scanner 125 microns
LPO 50 microns
HPO 12.5 microns
OIO 5 microns
1. Find the length
and width.

2. What is the
diameter?

250 microns and
100 microns

250 microns x 100
microns

Total Magnification: 400x
(HPO)
Objectives Size of 1 box
Scanner 125 microns
LPO 50 microns
HPO 12.5 microns
OIO 5 microns
1. Find the length
and width.

2. What is the
diameter?

250 microns x 100
microns

250 microns and
100 microns

Total Magnification: 1000x
(OIO)
Objectives Size of 1 box
Scanner 125 microns
LPO 50 microns
HPO 12.5 microns
OIO 5 microns
1. Find the length
and width.

2. What is the
diameter?

5 microns x 5
microns

5 microns and 5
microns

7. Calculate for the dimensions of the representative
microorganisms using the value of 1 OM and fill in Table 1.2.
Microorganisms Dimension
Amoeba 250 microns x 175 microns
Apergillus (sporangium) 12.5 microns x 12.5 microns
Bacillus 5 microns x 2 microns
Euglena 37.5 microns x 25 microns
8. When calibrating the ocular micrometer for use with the oil immersion
lens, you find that 10 SU coincided 60 OU.

(a) What is the length of each ocular unit?

X = no of boxes occupied by an organism
Y= no of lines occupied by an organism
OU = ocular units
SU = 10 microns (Constant)

X(OU) = Y(SU)

(OU) = Y(SU)/X


(OU) = 10(10)/60

(OU) = 1.67 microns
(b) What is the diameter and length of a microorganisms if its
diameter and length in OU are 2 and 20 respectively? (Show
calculations)

Given:
Length of each box (OU) = 1.67 microns
Diameter = 2 boxes
Length = 20 boxes


Diameter = (1.67 microns) (2 boxes)
= 3.34 microns

Length = (1.67 microns) (20 boxes)
= 33.4 microns





9. Identify the etched numbers in the
objective lenses







LPO HPO OIO
Focal length
(mm)
16 mm 4 mm 1.8 2.0 mm
Resolution
(microns)
1.1 microns 0.02 microns 6.22 microns
Magnification 10x 40x 100x
Numerical
Aperture
0.25 0.55 0.65 1.25 -1.4
Working
distance (mm)
4 8 mm 0.5 07 mm 0.1 mm
Conclusion

The microscope is a very powerful tool for understanding the
size, structure and function of different microorganisms that cannot
be seen with the unaided eye and that are not within the resolution
range of the normal eye. It uses different lenses to bend light to
reach the preferred magnification ranging from 40x 1000x
(compound microscope) understanding the capabilities and
limitation of a microscope is important if one is to get the best
results from a microscope.