LABORATORY

MICP

Bachelor of Science in Nursing: MICP LAB

LAB MODULE LAB UNIT WEEK

1 1 - PRELIM PERIOD 1

Basic Laboratory Equipment and Procedures in the Study of Bacteria

 Implement a study habit to read and comprehend the following prior to online class proper:
unit objectives; lab module and lab learning videos.
 Actively search for unfamiliar medical terminologies and relate to discussions.
 Establish effective teacher- student interactions through participation in the synchronous
online class discussion. through LMS discussion board or through Online Lab Class chat box. Ask
relevant questions.
 Answer and submit lab unit tasks online if there is any.
 For additional direction read study guide prior to class proper

At the end of this unit, the students are expected to:

1. Describe different Types of Microscopes

2. Identify and describe the basic laboratory equipment and functions.
3. Explain how to use and care for a compound light microscope.
4. Learn how to use a low power, high dry, and oil immersion lens to focus and observe specimens.
5. Learn to distinguish between prokaryotic and eukaryotic microorganisms.

MODULE OUTLINE
Basic Laboratory Equipment

 Types of Microscopes
 Equipment Used in Microbiology Laboratory

1
  Basic parts of a compound light microscope and their functions
 Proper care and handling of a compound light microscope
Procedures in the Study of bacteria
 Focus and view specimen using low power, high dry and oil immersion lens
Prokaryotic & eukaryotic microorganisms
 Prokaryotic and eukaryotic microorganisms

LAB GUIDE

This lab module was adapted from the book.

Microbiology and Parasitology. A Textbook and Laboratory

Manual for the Health Sciences, 2nd Edition

by Dr. Quiles and Dr. Bartolome. (C & E Publishing)

Read pages pages 4 through 10

Types of Microscopes

 In ancient Greek, ‘mikro’ means ‘of minute size’ and ‘skopion’ refers to ‘means of viewing’.
 Microscopes can be classified in a number of different ways; based on the source of light (light,
electron etc), arrangement, number of lenses (simple, compound), or method of interaction
between the sample and lens (probe, laser etc). Simple microscopes use the power of a single lens
to magnify a given sample. While compound microscopes use an objective lens to collect an image
enhanced by a secondary system of lenses.

Here are types of microscope, their specific qualities, and uses microscopes:
 Compound  A microscope that uses multiple lenses to magnify the image of a
microscope. sample. A compound microscope is used to examine materials at high
magnification (40-1000x), which is accomplished by combining the
effects of two lenses: the ocular lens (in the eyepiece) and the
objective lenses (close to the sample).
 The total magnification is derived by multiplying the ocular lens
magnification by the objective lens magnification.
 Brightfield  Also known as the Compound Light Microscope.
Microscope  It's an optical microscope that creates a dark image against a bright
background by using light rays. It is the most common microscope
used in biological, cellular, and microbiological laboratory research.
 Bright field microscopy is best suited to viewing stained or naturally
pigmented specimens such as stained prepared slides of tissue
sections or living photosynthetic organisms.
 Darkfield  Dark-field microscopy is perfect for illuminating unstained samples
microscope against a dark background, making them appear brightly lighted. A
special condenser in this type of microscope scatters light and causes
it to reflect off the material at an angle. The condenser is designed to
create a hollow cone of light instead of illuminating the sample with a
filled cone of light.
 It is more useful in examining external details, such as outlines, edges
 ideal for viewing objects that are unstained, transparent and absorb
little or no light.

 Phase contrast  Phase contrast is a light microscopy technique used to enhance the
Microscope contrast of images of transparent and colourless specimens. It
enables visualization of cells and cell components that would be
difficult to see using an ordinary light microscope.
 can also use dark field in the research of live bacterium, as well as
mounted cells & tissues.
 Differential  Known as Nomarski microscopy or imaging, differential interference
Interference contrast microscopy takes advantage of differences in the light
Contrast microscope refraction by different parts of living cells and transparent specimens
(DIC) and allows them to become visible during microscopic evaluation.
 type of imaging is that the three-dimensional

 Magnifying properties of a light microscope are merged with the

Fluorescence Microscope fluorescence emitting properties of molecules. Fluorescence
microscopy uses a high-intensity light source to trigger fluorescent
molecule called as fluorophore in the sample under investigation.
 generally used in combination with antibodies

Confocal Microscope  Known as confocal laser scanning microscopy (CLSM) or laser confocal
scanning microscopy (LCSM).
 an optical imaging technique for increase optical resolution &
contrast of a micrograph by means of using spatial pinhole to block
out-of-focus light in image formation
 uses lasers & fluorescence to create a 3 -dimensional image of a
sample
Electron Microscope  This is a microscope that uses a beam of accelerated electrons as a
source of illumination. As the wavelength ofan electron can be up to
100,000 times shorter than that of visible light photons, electron
microscopes have a higher resolving power than light microscopes
and can reveal the structure of smaller objects
 Scanning Electron Microscope (SEM) is a type of electron
microscope that produces images of a sample by scanning the surface
with a focused beam of electrons. The electrons interact with atoms in
the sample, producing various signals that contain information about
the surface topography and composition of the sample.
 Transmission electron microscopes (TEM) are microscopes that use a
particle beam of electrons to visualize specimens & generate ahighly-
magnified image. TEMs can magnify objects up to 2 million times
 The chief differences between the two are that the TEM gives a two-
dimensional picture of the interior of the sample while the SEM gives a
three-dimensional picture of the surface of the sample
Scanning Probe  Scanning probe microscopy is used to create images of nanoscale
Microscope surfaces and structures or manipulate atoms to move them in specific
patterns. It involves a physical probe that scans over the surface of a
specimen gathering data that is used to generate the image or
manipulate the atoms.
 forms images of surfaces using a physical probe that scans the
specimen.
 Scanned-probe microscopy (SPM) produces real-time images of highly
magnified, three-dimensional formed specimens. SPM uses a tiny
probe to scan the object's surface, bypassing the limitations of
electron and light microscopy. SPM refers to a range of related
technologies for imaging and measuring surfaces down to molecules
and atom groups on a micro scale.

Other Types of Microscope

Cryo-electron The device is a modification of transmission electron microscopes
Microscope (TEM). While TEMs use a beam of electrons to examine samples, most
biological materials degrade in these conditions.

This allows for high resolution of biomolecules by freezing them. The

device also uses electron beam which does not affect biological
structures thus enabling visualization of proteins, DNA and other bio-
molecules as they move and perform function.

In 2010, these devices enabled the visualization of atoms in a virus.

Researchers have used the cryo-electron microscopy to examine impact

of COVID-19 vaccine on human cells and found that it helps the cells
develop spike proteins similar to those found on the virus, which in turn
allows the body to build some immunity.
Acoustic Instead of using light, acoustic microscopes study samples using sound
microscope waves. This allows for non-invasive sample imaging. Ultra-sound is the
 most popular acoustic microscope modification.

Digital Microscopes
To perform live imaging, digital microscopes, which were initially
presented in 1986, use a digital camera and a computer. Some devices
have eyepieces, while others are entirely computer-controlled. The
computer can examine image aspects that are not visible to the naked
eye, such as distance measurements, fluorescence strength, and
minute thickness fluctuations.

Dino-Lite digital Dino-Lite digital microscopes are a recent innovation. These are
microscopes handheld devices, smaller than a pen that are capable of up to 500X
magnification.

Now Answer your LAB WOOKSHEET:

Pages 457 - 459.

in Microbiology andParasitology: Textbook and

laboratoryManual for the Health Science , 2nd Edition
by Bartolome & Quiles (C & E PUBLISHING)

Parts, Use Care Of a Microscope And Other Lab Equipment/Instruments

Because the microscope is such a valuable device, proper care and maintenance are essential.

1. Carry the microscope with both hands, one firmly clutching the arm and the other firmly grasping the
base. Avoid sudden jars.

2. Maintain the microscope and lens systems' cleanliness: Do not put your fingers on the lenses in any
way. If the lenses become dusty, clean them lightly using lens paper. Never leave a slide on the
microscope when it is not in use.

3. To keep the microscope from breaking, follow these steps: Never force the adjustments. All
adjustments should work freely and easily. Never allow an objective lens to get stuck in the slide or
cover slip, or even touch it.When viewing through the microscope, never use the coarse adjustment to
focus lower. Always keep your head to the side and your gaze parallel to the slide so that downward
movement is stopped before the objective reaches the slide. Never swap objectives or eyepieces
between microscopes, and never remove the front lenses from the objectives under any
circumstances.Never hold two microscopes at the same time.

4. Proper storage of the microscope:Make sure the immersion oil is no longer present in the
lens.Turn the stage all the way down and place the low power (10X) objective in place.Make sure the
slide holder does not extend past the microscope's left edge.The electrical cord should be wrapped
around the cord hanger.

For this, we will utilize the following interactive resources. CLICK HERE to access YouTube video:

Video Link

Video A Equipment Used in Microbiology https://www.youtube.com/watc

Laboratory/biotechnology lab equipment h?v=8qIMVbWPoXc&t=1s
/Instrument formicrobio lab

Video B Parts & Functions of Microscope https://www.youtube.com/watc

h?v=l4Lz73OMWkM

Video C Care And Use Of The Microscope https://www.youtube.com/watc

h?v=t2ncnZH_YFI

Low Power, High Dry, And Oil Immersion Lens

The light microscope uses a variety of glass lenses to produce a magnified image that is focused before
the eye. the resolution of the light microscope is sufficient to produce excellent images of many of the
important cell structures and organelles

Many of the diseases that have plagued humanity have been discovered at the microscopic level. The
microscope, one of humanity's greatest inventions, has aided in the discovery of causes and, as a result,
remedies for many of these diseases.

Let us now learn how to focus and view specimen using low power, high dry and oil immersion lens. For
this, we will utilize the following interactive resources. CLICK HERE to access YouTube video:
Video Link

Video D How to Focus a Microscope & How the Field of View https://www.youtube.com/watc
Changes h?v=zzamomqlwxU
 Image:MDSmafe fb

Prokaryotes And The Eukaryotes

Some cells are uncomplicated, but others are complex with internal structures called organelles. These
cells can be put into two categories which are the prokaryotes and the eukaryotes, and they have many
differences and similarities. Humans and other multicellular organisms are eukaryotes while Prokaryotic
cells comprise bacteria and archaea. What is the difference between prokaryotic and eukaryotic
microorganisms?

Let us now compare and contrast prokaryotic and eukaryotic cells, their structure. For this, we will
utilize this learning resource.

CLICK HERE to access YouTube video:

Video Link

Video E Prokaryotic Vs. Eukaryotic Cells https://www.youtube.com/watch

?v=RQ-SMCmWB1s
 LAB TASK

Direction: After your instructor reviewed the parts and functions of the compound light
microscopes and other instruments of a micro lab., fill in the table on your workbook

Identify the parts of a compound light microscope, Cite its function. USE YOUR OFFICIAL
LAB WORKSHEET: Pages 457 - 459. in Microbiology andParasitology: Textbook and
laboratoryManual for the Health Science , 2nd Edition by Bartolome & Quiles (C & E
PUBLISHING)

Class discussion: Comprehension and Knowledge.

1. Describe the major differences between light microscopes and electron microscopes

2. Describe how to clean and maintain your microscope, what you will use to clean the
microscope's lens. Things you will need to do after you are done with the microscope at the
end of the lab.

3.Identify and describe the basic laboratory equipment and functions.

A. B. C.

Image from LearningInHand.com

 D. E. F.

. E. F.

4. Distinguish between prokaryotic and eukaryotic microorganisms and compare them usingthe
characteristics listed below:
Prokaryotic Both (Prokaryotic And Eukaryotic) Eukaryotic

When viewed under microscopes :

Which lacks a cell nucleus ?
Which contains many different structures?
The Bacteria is classified as?
Which is a single-celled organism?
Which is Multicellular?
Which possesses a membrane-bound nucleus?
Which contains genetic materials?
Which is a cell?
Which contains ribosomes and cytoplasm?
The Fungi is Classified as?
 Congratulations! You reached this far.
Here are some terminologies used in this module:

TERMINOLOGIES

 Field Of View. The field of view refers to how much you can see through the eyepiece or on the
screen (FOV).
 Immersion Oil. A special technique used in light microscopy that involves the immersion of both
the specimen and the objective lens in a transparent oil that has a high refractive index.
 Internal Structure. The inner pieces and parts that keep organisms alive, help them grow, and
help them reproduce.
 Magnification. The amount by which the dimensions of an image are, or appear to be, magnified
when compared to the same dimensions in the actual sample is known as magnification.
 Optical microscopes. Refers to an optical instrument that uses a lens or an arrangement of lenses to
magnify an object.
 Organelles. Cell organelles are specialized entities present inside a particular type of cell that
performs a specific function.
 Resolution.The ability to see small, close-together things as distinct is referred to as resolution.
REFERENCES

Bartolome and Quiles. (2020) Microbiology and Parasitology: A Textbook and Laboratory
Manual for the Health Sciences. 2nd Edition. C&E Publishing House. Quezon City. QR65. B37
2020

Engelkirk, Paul G. (2019). Burton's microbiology for the health sciences, 8th ed. . Philadelphia :
LWW.616.01 E3 2007

Tortora, G.J. (2014) Microbiology. Pearson: Singapore. 616.9041 T63 2014

www.cdc.com

www.googlesearch

Prepared by: Aida V. Garcia, MAN, RN for MICP