BIO210L Manual - Part One - Spring 22-23

BIO210L
Cells and Molecules Laboratory

Course Packet
Spring 2022-2023
Table of Contents
Introduction: Guidelines for Writing a Lab Report ................................................................................................ 3

Introduction, Rules, and Regulations................................................................................................. 11
Experiment 1: Microscopes and Microscopy .................................................................................... 15
Experiment 2: Microscope Anatomy, Use, and Care ........................................................................ 23
Experiment 3: Cell Staining............................................................................................................... 29
Experiment 4: Miscellaneous Staining and Cell Diversity ................................................................ 34
Copyright © - AUST 22-23

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Introduction: Guidelines for Writing a Lab Report
The guidelines below will drive you through the process of writing a lab report. Make sure to read
those guidelines properly and incorporate them into your report.
A- Important notes
- The reports are to be presented in a descriptive format. Assume that the reader has little
background regarding specifics of your experiment, but has a general knowledge and interest in the
subject matter. The format should be similar to basic scientific papers.
- When writing a report, it is utterly forbidden to copy (plagiarize) any type of information from
any type of source, including the lab manual. Plagiarism consists in copying any content from an
information source, whether a book, a website, a teaching manual, or even thesis reports written by
classmates or colleagues. Even copying a short sentence from a resource is considered as
plagiarism. Plagiarism will lead to a zero on the lab report.
- When using an information resource, you have to keep the scientific meaning of it but you have to
rephrase using you own words. You are not allowed to copy the information as it is provided
by the source or it will be considered as plagiarism.
- Never use slang and conversational English expressions. Those are not tolerated in formal writing.
For instance, you are not allowed to write “u” instead of “you”.
- Your writing should be in full sentences and easily understood. It should conform to the
conventions of standard written English (sentence form, grammar, and spelling). One reason for
emphasizing clarity is that writing and thinking are closely related.
B- The content of the lab report

The lab report contains a cover page and the following seven sections:
- Introduction (with the objectives of the experiment stated at the end of the introduction)
- Materials (divided into” Equipment” and “Reagents”)
- Procedure
- Results
- Discussion
- Acknowledgements
- Literature (alphabetical by author)
B.1. Cover page

The cover page contains the title of the report, your name, the date, and the name of your laboratory
instructor. See below the format of the cover page.

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American University of Science and Technology
(Size: 18, Font: Times New Roman)(One line, Line spacing = “Single”)
Faculty of Health Sciences
(Size: 14, Font: Times New Roman) (One line, Line spacing = “Single”)
Department of Laboratory Science and Technology
(Size: 14, Font: Times New Roman)
(9 lines, Line spacing = “Single”)

BIO210L: Cells and Molecules Laboratory

Experiment I
Bile Salts Enhance Lipase-Digestion of Fats
Name: Your Name

Instructor’s Name: Dr. XXX (Size: 14, Font: Times New Roman)
Date Experiment Done: October 20, 2013

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B.2. Introduction
The report begins with an introduction; it is a brief background about the topic that was covered
during the lab session. The introduction provides information from the more general to the more
specific.
1- Focus the introduction on the major aspects of the lab topic. For instance, if the laboratory
session is about DNA sequencing, then your main highlights of the introduction would be to
explain DNA structure, the main techniques of sequencing, the technique specifically used
during the lab session, and its applications in the field of biology.
2- Do not include any information that goes beyond the scope of the lab topic: be concise,
brief, and read the sentences at least twice to make sure you are not including useless or
repetitive information or words.
3- Introduction should be no longer than 350 words and split into 2 to 3 paragraphs (one
paragraph per aspect described).
4- Use the present and the past tenses depending on the phrasing.
5- Do not include tables, bullets, lists, figures or any such subdivisions in an introduction (unless
specifically requested by the instructor). Introduction should be written in paragraphs.
6- The introduction should never start on the cover page, it always starts on a new page.
7- The objectives of the lab session should be included at the end of the introduction. Objectives
describe the exact purpose of the lab session. For instance, if the lab session is related to DNA
sequencing, then the aim of the lab session would be written as follows:
“More specifically, the purpose of the lab session was to make a real time sequencing of a 1kb
fragment of a DNA extracted from human skin tissue”.
B.3. Materials
1- The materials section is divided into “Equipment” (plastic ware and glassware) and “Reagents”
(chemical and biological reagents).
2- You can make a table or a list for each of the equipment and reagents.
3- For each equipment used, specify the exact name and brand of it (e.g., “sequencer machine from
Applied biosystems reference AB120”). For each glassware or plastic ware used, specify their
capacity (“1.5ml Eppendorf tubes” not “tubes” simply).
4- For each reagent used, specify the concentration of the stock solution you have used.
5- Do not include in this section the functions of each material used, this information will be
provided in the discussion section.

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B.4. Procedure
1- The aim of this section is to describe the full procedure used during the experiment by
listing the steps performed in order. Anyone who reads your report should be able to
perform the experiment exactly the way you did. Do not include results here.
2- You can number the steps or use bullets. Do not write the procedure as one bulk paragraph.
3- For each step of the procedure, specify the machine used, the reagents used, reaction time,
and parameters used.
4- For the reagents used, you specify the volume and the concentration of the stock solution
you have used.
B.5. Results
In this section you write down the exact results you have obtained and observed during the
experiment. Clearly phrase in your own words the results you obtained. Do not leave a result up to
the interpretation of your instructor. Use figures whenever relevant to the comprehension of your
results; make sure to add a number to every figure, as well as a legend.
1- Use the past tense when describing your results.
2- Use the passive voice when describing your results.
3- In this section do not discuss the results. Only give an objective description of the results
you obtained.
4- Tables and figures are numbered and described using a one-sentence legend.
5- Do not include figures unless they are of interest to explain the results and use figures of
decent dimensions (make sure the figure does not cover the entire page).
B.6- Discussion
In this section make conclusions based on your results, data, and personal observations.
1- Use the past tense and present tense. Use the past tense when referring to something you did in
the lab. Use the present tense when referring to known facts relevant to your results.
For example, results should be reported in the past tense ("The solution turned blue..." or "The
optical density of the culture doubled in 20 minutes..."). Established facts can be reported in the
present tense ("Logarithmic growth occurs in bacteria following..." or "Restriction
endonucleases cleave double-stranded DNA..."). Be consistent in a given section with the use of
tense, avoid switching back and forth from past to present tenses.
2- Do not use the personal forms “We, us, ours...”.
3- For each step, specify the role(s) of the reagents used. Be brief and concise.
4- Organize your discussion from the more general to the more specific.
5- Whether you obtained positive or negative results, make an interpretation of those results.
6- In case of negative results, troubleshooting is a must. Go back to the materials and equipment
used, as well as the steps of the procedure to make interpretations of what could be the reason
for those results. Most importantly, search the literature to discuss the results. Do not limit
yourself to the content of the lab manual.

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B.7. Acknowledgments
Acknowledge those who assisted you in any way.
B.8. Literature
Lastly, include a literature section that lists all the resources you have used to organize the content
of the lab report. These citations refer readers to a complete list of references at the end of your
report.
1- List your resources in alphabetical order using the last name.
2- Literature citations are generally used in the introduction and the discussion. They provide
more background and support any conclusions made in your report.
3- Resources can be books, websites, lab manuals, articles, or reviews.
C- Template for a lab report

Below is a template of a laboratory report, refer to it while writing your own. Make sure to follow
all instructions closely. In this example of a lab report, the purpose of the lab session was to test for
the effect of the absence/presence of bile salts on the efficiency of milk digestion by a pancreatic
lipase. This is how a report should be written:

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BIO 210L (Size: 8, Font: Times New Roman) BIO 210L Bile Salts Enhance Lipase-Digestion of Fats
Introduction
The pancreas secretes various enzymes into the small intestine. One of these enzymes, pancreatic
lipase, digests dietary fats into products such as glycerol and fatty acids. However, fat is insoluble
in water-based chyme (liquefied food processed by the stomach); and in the intestines the fats cling
together, providing little surface area for attachment of the enzymes. This prolongs the time it takes
the lipase to digest the fat.
In order to speed up the fat digestion process, bile salts, secreted by the gall bladder into the small
intestine, act as a detergent which break up the fat droplets in the watery chyme thus increasing the
surface area for enzymatic digestion by lipase. In other words, bile is an emulsifying agent.
Emulsification of fats is achieved upon exposure to bile salts, which allow pancreatic lipase to
digest the fat more efficiently. In order to demonstrate that bile salts enhance the digestion of fats,
the digestion of milk fat was tested by pancreatic lipase in the presence and absence of bile salts.
Materials
a- Equipment
- Eppendorf plastic tubes 1.5ml and 2ml
- Centrifuge (Eppendorf 13.0)
- pH meter
b- Reagents
- Dehydrated pancreatin reconstituted in water at 1 g/100ml
- Dried grains of bile salts
Procedure
1. Three groups of test tubes were set up; 3 replicates were set up in each group in order to provide
an adequate sample size.
2. 3 plastic test tubes for each of the 3 groups tested were placed in a rack and labeled as follows:
1.1-1.3 for group 1, 2.1-2.3 for group 2, and 3.1-3.3 for group 3.
3. To each group of 3 test tubes, the following reagents were added in the following order
usingplastic droppers:
Group 1: 3ml of whole milk + 5ml of water + 3 grains of bile salts
Group 2: 3ml of whole milk + 5ml of pancreatin solution
Group 3: 3ml of whole milk + 5ml of pancreatin solution + 3 grains of bile salts
4. Tubes were incubated at 3° C for 1 hour. During that hour, the pH was tested every 20 min
using a pH meter and the first measure was determined at time zero (beginning of the
experiment).
5. Values were then organized in a chart for each of the time points: the average pH was
calculatedfor each set of replicates and the 4 time point measurements of the pH were specified.

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Results
The pH did not decrease during the 60-minute incubation period in the negative control group
#1which only contained milk and bile salts (Table 1). In groups #2 and #3 the pH did decrease as
the digestion of fats progressed and fatty acids built up in the test tubes.
After 20 minutes the pH decreased in group 2 from 8.5 to 7.5, while there was a greater change in
tube 3 (from pH 8.5 to7.0). At 40 min incubation, the pH of the solutions in both groups #2 and #3
had dropped to 6.5 and did not decrease further at 60 minutes. The data for all 3 groups is visually
plotted in Figure 1.
Table I: Mean* pH of whole milk during incubation with bile salts and/or pancreatin.
Time (min.) Group 1 (+B.S.1) Group 2 (+Pancreatin) Group 3 (+B.S.+ Pancreatin)

0 8.3 8.5 8.5
20 8.4 7.5 7.0
40 8.3 6.5 6.5
60 8.3 6.5 6.5
* mean pH of 3 sample tubes per group

1
Bile Sales
Discussion
The dehydrated pancreatin was reconstituted in water immediately before using. This solution
contains the pancreatic lipase enzyme which is used to digest the milk fats. The negative control
tube which only contained bile salts and milk, did not exhibit a pH change, which a result that is
expected. Therefore, it was concluded that bile salts alone did not digest milk fats. Moreover, it was
observed that as fatty acid concentration increases, pH decreases. Digestion of milk fats occurred in
tubes 2 and 3, based on the observation of fatty acid by-product accumulation, as measured by a
decrease in pH. The production of fatty acids occurred faster in group #3 as evidenced by the data
at 20 minutes. This suggests that an enzyme present in the solution of group #3 aided in the

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digestion of the fats. Since the concentration of pancreatin was identical in both groups #2 and #3,
the addition bile salts must have contributed to the digestion of the fats by breaking up those fat
droplets into smaller particles which increased the availability of substrate in
group #3. The action of bile salts enhanced the rate of the lipase-fat digestion. The data from tube
#1, as a negative control, demonstrates that bile salts do not digest fats; therefore, bile salts must
only act to emulsify the fats thus enabling pancreatic lipase to act more efficiently. This conclusion
is also supported by work done by Patton and Carey (1979) and reported in an article by Bowen
(1996).
It was also determined that either the enzymatic activity of pancreatic lipase is inhibited below
pH6.5, or that the fat substrates were depleted by 40 minutes in tubes 2 and 3, since there was no
change in pH between 40 and 60 minutes. Further experimentation will reveal which of these two
possibilities occurred. Fox (1996) reported that the optimal activity of pancreatic lipase occurs at a
pH of 8. Therefore, it is most likely that the lack of change in pH between 40 and 60 minutes in
tubes 2 and 3 was due to the fact that the accumulation of fatty acid end products produced an
excessively acidic environment (pH 6.5 at 40 minutes), thereby inhibiting further enzyme activity.
This may suggest that pancreatic lipase is denatured in weakly acidic conditions between pH 7.0and
6.5. However, if this is not the case, then an alternative conclusion may be that depletion of
substrate occurred during the experimental period. This experiment should be re-run using cream or
vegetable oil which contain significantly more fat than whole milk.
Acknowledgments
I thank my instructor for her help with the t-test and my lab partners for helping me understand this
experiment.
Literature
- (APA format must be used here) Bowen, R. 1996. Absorption of Lipids. Website at
URL:http\\www.arbl.cvmbs.colostate.edu/hbooks/pathphys/digestion/smallgut/absorp_lipids.html.
- Fox, S.I. 1996. Lab Guide - Human Physiology, Concepts and Clinical Applications. 7th ed. W.C.
Brown, Dubuque, IA.
- Mader, S.S. 1995. Human Biology, 4th ed. W.C. Brown, Dubuque, IA.
- Martini, F.H. and M.J. Timmons. 1997. Human Anatomy. 2nd ed. Prentice Hall, Upper Saddle
River, NJ.
- Patton, J.S., Carey, M.C. 1979. Watching fat digestion. Science 204: 145.

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Introduction, Rules, and Regulations

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Responsible behavior in a lab aims at protecting yourself as well as your colleagues from any
contamination, intoxication, or injury. Irresponsible behavior in a lab is not tolerated as it puts other
people’s health at risk and might lead to severe consequences.
A- General Biology Lab Safety Rules:
1. Open-toed shoes are not permitted.

2. Smoking, drinking, and eating are totally prohibited.
3. Never wear loose jewelry while working in a laboratory.
4. Personal items unrelated to the lab work are forbidden in the lab. Instead, use your locker to
store your personal items.
5. Before entering the laboratory, cover any cut on your hands with a bandage.
6. Make sure to tie your hair.
7. Some chemicals used in the laboratory are absorbed through contact lenses. It is advisable to
remove contacts before lab sessions or wear tight fitting goggles when appropriate.
B- While Working in the Lab:
1. Always wash your hands with soap before and after each lab session.
2. Gloves must be worn throughout the experimental work.
3. Benches must be cleaned before and after each laboratory session using 70% EtOH prepared in
distilled water.
4. Do not touch any equipment or area with contaminated gloves, especially when using ethidium
bromide.
5. Wear eye or face protection when using the transilluminator.
6. Safety goggles must be worn in the lab when instructed.
7. Use the laboratory equipment properly, following the manufacturer’s instructions. Do not use
any equipment unless you have been made aware of its manipulation and safety procedure. If
you do not know how to use a device, ask for your instructor’s or lab assistant’s help.
8. At the end of the lab session, Glassware and instruments must be cleaned with tap water,
then distilled water, and placed on the bench next to the sink for air drying.
9. Other materials that do not require any cleaning must be returned to their proper storage area.
10. No lab material of any kind may leave the laboratory.
C- Manipulation of Chemicals:
1. Whenever using a chemical, make sure to read the pictogram displayed on the container, box,
bottle, or tube.
2. Always wear gloves when manipulating any bottles, even if properly sealed:
- Never hold a container/tube from its cap or neck, always grab tightly the bottle itself.
Copyright © - AUST 22-23 Page 12

- Never carry more than one container in one hand.

- Make sure the bottle/container/tube is properly sealed when carried from one bench to another.
- When you are done with using a chemical, immediately seal it. Always make sure to
close bottles/containers/tubes properly.
3. Never pipette out solutions with your mouth: use the mechanical or electric pipettor.
4. After you use any chemical, enzyme, or solvent, return it to its storage place (fridge, freezer,
orbench shelf).
5. Dispose of all chemical waste in the proper waste container. Never pour any chemical into the
sink without permission from your instructor.
D- Safety Procedures Related to the Manipulation of Equipment:
1. Care must be taken when handling and carrying microscopes. Always carry microscopes with
both hands and drop off slowly on the bench.
2. Never push a microscope on any surface: vibrations severely alter the mechanism and the lenses.
3. When putting away a microscope:
- Gently clean the lens with the appropriate soft wipes.
- Reduce light intensity and then switch of the lamp. Never switch of the lamp without
reducingthe light intensity first.
- Set the nosepiece to the lowest magnification level.
- Clean the immersion oil from the lenses and the stage.
- Unplug the electric cable when you finish using the microscope.
4. Make sure that gas nozzles and water faucets are switched off before leaving the lab.
5. Be cautious when using hot plates: make sure that everyone in the lab is aware of the plate
being hot, especially after use. Leave a note and do not carry or move until temperature goes
down.
6. Tie back hair when using Bunsen burners and be extremely cautious when working next to the
flame. Do not place any flammable chemicals around the burner.
7. When using a centrifuge, make sure to balance the rotor by placing the tubes/falcons in facing
slots. Make sure that the centrifuge is not noisy otherwise it indicates imbalance and the rotor
might break. In such case stop the centrifuge immediately or unplug it.
8. Care must be taken when working with models. All models must be reassembled (or
disassembled when working with molecular models) and returned to their proper storage area
at the end of each lab period.
9. Chairs should be pushed under the benches at the end of the lab session.

E- Biohazard and Chemical Waste Disposal:
1. Always dispose of chemical waste in the allocated storage containers.

2. Never pour any chemical into the sink unless indicated by your instructor.
3. Solid biohazard waste is systematically collected in the red-bag lined biohazard waste bags.
4. Dispose of broken glass into the Glass Disposal Box.
5. Dispose of slides, toothpicks, and any other small sharp object, whether contaminated or not
with biological tissue, into the sharps’ container.
6. Even though simulated blood and sterile blood used in some experiments are considered non-
hazardous, it should be treated as such. Dispose of any material that has contacted human or
animal tissue into the Biohazard Waste Box.
F- In Case of Accidents in the Lab:

1. In case of accidents related to broken material or liquid spill on the floor or bench, notify your
instructor immediately. Do not clean up broken glass or spilled chemicals yourself.
2. If any toxic chemical, antibiotic, or carcinogen is spilled on your skin or cloth, immediately
remove the contaminated cloth and wash skin area thoroughly under tap water for at least 5
minutes. Spills on legs can be rinsed in the sink. Do not hesitate to use the lab shower in the
laboratory on large spills. Do not rinse contaminated area with alcohol.

Experiment 1: Microscopes and Microscopy

A- Introduction
The microscope is an instrument used to observe magnified images of small objects difficult to
observe with the naked eye. It is derived from the Greek word "micro" (tiny) and "scope" (to view).
Microscopy is the science of investigating small objects using microscopes.
Microscopes are fragile instruments. Users must avoid damaging the devices by following a strict
procedure. Carefully bring a microscope to your workspace as per the below instructions:
* Always carry microscopes with both hands, one supporting the base from below, and the other
gripping the arm of the microscope
* Place the microscope gently on a table, a bench, or any horizontal surface.
* Never push a microscope on any surface: vibrations severely alter the mechanism and the lenses
B- Major types of microscopes

One of the most important tools used by biologists is the microscope. It is an exquisitely sensitive
arrangement of lenses that, when appropriately used, provides extremely accurate imaging of both
live and fixed tissues.
a- Light microscope:
The light microscope is the basic tool for cell biologists, with technical improvements allowing the
visualization of ever-increasing details of cell structure. The light microscope makes use of visible
light.
Contemporary light microscopes magnify objects up to about a thousand times (1000x) their size.
Most cells are between 1 and 100 µm in diameter and can be observed using light microscopy to
identify larger sub-cellular organelles, such as nuclei, chloroplasts, and mitochondria. However, the
light microscope is not powerful enough to reveal fine details of cell sub-structures. In such cases,
resolution (the minimal distance required to distinguish two points) becomes more important
than magnification. Images can be magnified as much as desired, but such magnification does not
increase the level of detail that can be observed. The limit of resolution of the light microscope is
approximately 0.2 µm; two objects separated by less than this distance appear as a single image,
rather than being distinguished from one another.
A light microscope is called “simple” when it has a single objective or “compound” when it has
several objectives. The three major types of light microscopes are: the brightfield, the darkfield,
and the phase-contrast:
* The brightfield microscope produces an image where the object appears dark on a bright
background. The object appears dark due to the absorbance of the visible light by the object. Often,
more detailed images can be obtained by adding a staining reagent to the tissue observed (cells,
sections of organs, or other) that is being observed.
* In the dark-field microscope, the light passes directly through the cell. Ability to distinguish
different components of the cell depends on the contrast resulting from the absorption of visible
light by the cell components. This microscope contains a special condenser that directs the light
into the sample. Only the light scattered from the sample will enter the objective lens and produce
an image. This image produces a light object on a dark background.
* An alternative microscope is the phase-contrast microscope, in which small phase shifts in the
light passing through a transparent specimen are converted into amplitude or contrast changes in

the image. It uses optical systems that convert variations in density or thickness between different
parts of the cell to differences in contrast that can be seen in the final image. Live, unstained
organisms, are seen clearly with this microscope, as well as internal cell parts such as mitochondria,
lysosomes, and the Golgi apparatus.

Refer to the below figure that displays the anatomy of a light microscope
There are slight variations among different microscope models and brands, but they are all similar
in basic structure.
Fig.1. Anatomy of a compound light microscope.

b- Fluorescence microscope
The fluorescence microscope is widely used because it is a sensitive method for studying the
intracellular distribution of molecules. A fluorescent dye is
used to label the molecules of interest, usually proteins,
within live or fixed cells. The fluorescent dye is a molecule
that absorbs light at a given wavelength and emits light at
another wavelength. The specimen is illuminated at a
wavelength of light that excites the fluorescent dye, thus
emitting a light that is detected using appropriate filters. One
frequent application is to label antibodies with fluorescent
dyes and target those antibodies against a specific protein and
detect the distribution of that protein within the cell. The
most widely used fluorescent dye is the green fluorescent
protein (GFP) isolated from the jellyfish. Fig. 2. Fluorescent GFP-Positive Cells
c- Confocal microscope
Confocal microscopy allows images of increased contrast and detail
to be obtained by analyzing fluorescence from only a single point in
the specimen. A small point of light, usually supplied by a laser, is
focused on the specimen at a particular depth. The emitted
fluorescence light is then collected by a detector, such as a video
camera. Before the emitted light reaches the detector, however, it must
pass through a pin-hole aperture (called a confocal aperture) placed at
precisely the point where light emitted from the chosen depth of the
specimen comes to a focus. Consequently, only light emitted from
the plane of focus can reach the detector. Scanning across the
specimen generates a two-dimensional image of the plane of Fig. 3. Different
focus, a much sharper image than that obtained with standard Fluorescent Staining in
fluorescence microscopy. Moreover, a series of images obtained at blood cancer cell
different depths can be used to reconstruct a three-dimensional image
of the sample.
d- Inverted microscope
An inverted microscope is a microscope with both the light source and condenser located on top
(above the stage and pointing down), while the objectives and turret are located at the bottom part
(below the stage and pointing up). Inverted microscopes are useful for observing live cells or
organisms located at the bottom of a container (e.g. cells in a tissue culture flask) under more
natural conditions than on a glass slide, as is the case with a conventional microscope.
e- Stereomicroscope
Stereomicroscopes, also called dissecting microscopes, are actually two compound microscopes
that focus on the same point from slightly different angles. This allows the specimen to be viewed
in three dimensions. As opposed to compound microscopes, the image is upright and laterally
correct (not upside down and backwards). Stereomicroscopes have a relatively low power of
magnification, usually below 100x. One can also manipulate the tissue while observing it, such as
performing surgeries (hence the name "dissecting microscope").

f- Electron Microscope
The energy source used in the electron microscope is a beam of electrons. Since the beam has an
exceptionally short wavelength, it strikes most objects in its path and increases the resolution of the
microscope significantly. The electrons travel in a vacuum to avoid
contact with deflecting air molecules, and magnets focus the beam
on the object to be viewed. An image is created on a monitor and
directly viewed on a screen. Electron microscopy is the tool of
choice to observe extremely small structures such as viruses or sub-
compartments of a cell.
The most traditional form of electron microscope is the transmission
electron microscope (TEM). To use this instrument, ultrathin slices
of microorganisms or viruses are placed on a wire grid and then
stained with gold or palladium before viewing. The densely coated
parts of the specimen deflect the electron beam, and both dark and Fig. 4. TEM of blood cancer cells.
light areas show up on the image.
The scanning electron microscope (SEM) is the most contemporary form of electron microscope.
Although this microscope gives lower magnifications than the TEM, the SEM permits three-
dimensional views of microorganisms and other objects. Whole objects are used, with gold or
palladium staining.
Fig.5. SEM of blood cancer cells.

Experimental Part:
1. Place the microscope with the ocular lens (or eyepiece) pointing toward you. The ocular lens,
which has a magnification power indicated on it, is the tube you look through to observe the
magnified image of the specimen. The ocular lens may have a movable pointer incorporated in
it. Do not remove the eyepiece from the body tube.
2. Locate the arm of the microscope. It is the upper body structure that supports the eyepiece and
the assembly of lenses. Grasp the microscope from both its arm and base when carrying it.
3. The objectives are essentially tubes that contain lenses of various magnification powers. The
magnification power is engraved or painted on each objective and is indicated by the number
located before the “x” symbol. The shortest objective has a magnification power of 4x
(pronounced “four times”, meaning four times the actual diameter of the specimen). The 4x
objective is sometimes called the scanning lens because its lower magnification gives an
overall view of the specimen. The next longer objective is the low power 10x objective. The
third objective known as high power may be either 43x or 45x. Some microscopes may have a
fourth objective, which is the longest. It is an oil immersion lens that has a magnification of
100x. An oil immersion objective requires that immersion oil is applied on the slide, in a way
that the objective lens becomes immersed in the oil once placed on top of the slide. The
immersion oil increases the resolving power since it has an index of refraction that is similar to
the glass index. Oil immersion will not be used during this lab session as it requires some
practice.
The total magnification of a specimen is determined by multiplying the power of the objective
by the power of the ocular lens. The ocular lens typically has a magnification power of 10x.
With the scanning lens in position, a specimen would be magnified a total of 4x10, or 40x.
The objectives are supported by the nosepiece. Change objectives by turning the nosepiece
right or left until you feel the next objective clicks into place and points straight down.
4. The stage is the horizontal tray that supports the specimen to be viewed. The specimen is
always placed on a microscope slide. For observation, the specimen must always be positioned
on top of the opening that is located at the center of the stage. Most microscopes are usually
equipped with stage clips that hold the slide in place by slipping it under the clips. Before
observing a slide, wipe it with 70% EtOH using the appropriate tissue.
5. The condenser is below the stage. It collects and condenses light coming from below into a
focused beam and shines through the hole and specimen on the stage. Light then passes
through the objective and other internal lenses forming a magnified image visible through the
ocular.
6. The Iris diaphragm associated with the condenser functions similarly to the iris of the eye, i.e.,
it regulates the amount of light passing through the microscope lenses, producing various
degrees of image brightness. Since many tissue specimens are nearly transparent, there may be
a need to reduce the light level to observe structures. Your microscope may have a small level
below the stage that moves to open and close the iris diaphragm. The iris diaphragm may also
be a rotating wheel with various size holes that allow more or less light to pass through the
specimen. Determine which kind of iris diaphragm your microscope has.
7. Focusing knobs are located on both sides of the microscope. The larger knob next to the body
of the scope is the coarse adjustment knob used for general focus upon the specimen. The
smaller knob is the fine adjustment knob used for focusing on small details.

8. The illuminator is located directly below the condenser. It contains a light bulb, which is the
light source of your microscope. The on-off switch may be just behind the illuminator, or it may
be found on the electrical cord.
Some microscope models have below the condenser a small circular mirror that can be adjusted
to reflect light into the condenser.
9. The base is the lower foundation of the microscope, and it sits on the table.

Experiment 2: Microscope Anatomy, Use, and Care

A- Introduction
The purpose of this session is to become familiar with the practical use of the brightfield
microscope and learn how to use its features in order to optimize the quality of the image observed.
B- Practical use of the microscope

Whenever using a microscope, one has to make sure that the below is strictly applied.
When putting a microscope away:
* Remove the slide from the holder.
* Gently wipe the lens with the appropriate soft wipes. Clean the lenses by gently wiping them in a
circular motion of the lens paper. If the lens is still soiled, notify your instructor.
* Reduce light intensity and then switch off the lamp. Never switch off the lamp without reducing
the light intensity first.
* Bring down the stage to its lowest level by using the coarse knob.
* Set the nosepiece to the lowest magnification level. The short 4x objective should always be in
place (pointing straight down) when the microscope is stored. This reduces the risk of hitting an
objective on a slide when starting a focus.
* In case oil immersion was used, wipe the 100x objective with a soft wipe and 70% EtOH.
* Unplug the electric cable and wind it around the base.
* Cover the microscope with a dust cover.

Experiment 1: Adjusting the focus and determining the image orientation.
Purpose
To become familiar with adjusting the focus on a specimen and
observe how the image produced by the light microscope is inverted.
A slide with a letter “e” already mounted on it will be used.
Materials Required
- Compound brightfield microscope
- Slide with a letter “e” mounted on it (slide is ready to use)
Procedure
1. Place the microscope (do not push it!) with the ocular pointing toward you. If the microscope
was properly stored, lowest objective should already be in position and stage should be at its
lowest level.
2. Plug in the electrical cord, turn on the illuminator, increase the light intensity.
3. Place the slide on the stage so that the letter is positioned right side up and centered over the
hole. Place the stage clips over the ends of the slide to hold it in position.
4. While looking through the objective, slowly turn the coarse adjustment knob clockwise to bring
the stage closer to the objective. Stop turning the knob when the “e” letter is in focus. Never
force an adjustment knob after it stops turning. The fine adjustment knob is usually not required at
low magnifications. At higher magnifications, always adjust the coarse adjustment knob first and then
the fine adjustment knob. The space between the slide and the objective is called working
distance. The field of view is the area of the slide or specimen that you can see through the ocular lens.
If you wear glasses; you might find microscopy observations more convenient without them.
5. Look at the magnified image of the letter.

What is the orientation of the “e” letter in the image in comparison to its position on the slide?
6. While looking into the ocular lens, gently push the letter slide to the left.
Which direction does the letter move?
7. Change the position of the iris diaphragm to produce various degrees of brightness, and then
adjust it to what is comfortable for your eyes.
8. Change objectives to the next higher power and adjust the focus. Now note the working distance
between the objective and slide. Did it decrease or increase?
9. Center the letter “e” within the field of view. Can you see the entire letter within the field?
10. Without moving the slide, use the next higher power objective and focus on the letter using both
adjustment knobs. Can you see as much of the letter as you did under the previous objective?
11. Change to the next higher power and focus on the letter using the adjustment knobs. Can you
see as much of the letter as you did when using the previous objective?
12. Move to the lowest power objective into position and focus on the letter “e”. Note the
brightness of the field. Now change to the high power objective and note the brightness of the
field. Is the field of view brighter under lower or higher power?

This is an important point to remember when using high power; the field becomes dimmer
(darker) with each increase in magnification. It is advised to use stronger light under high
power by adjusting the iris diaphragm. However, too much light with high power can wash out
fine details in the image.
13. Bring down the stage, remove the letter “e” slide from your microscope and return it to its
allocated place.
Experiment 2: The concept of depth of field.
Purpose
Become familiar with adjusting the focus on a specimen and observe how the image produced by
the light microscope is inverted. A slide with crossed-colored threads already mounted on it will be
used.
Depth of field is another characteristic of microscopy that changes with magnification. Depth of
field is vertical depth of an image that remains in sharp focus under a particular magnification. As
an example, a magnified image with great depth of field allows the viewer to observe several layers
of cells on a slide, all appearing in clear focus. A magnified image with lesser depth of field forces
the viewer to focus up and down through the cells, in order to see each layer in clear focus.
Materials Required
- Slide with crossed-colored threads (slide is ready to use)
Procedure
1. Place the slide that contains the threads on the stage and adjust the clips.
2. Using the scanning objective (4x), move the stage until the threads intersect at the center of the
field of view. Observe the three threads in the area where they intersect.
Are all three threads equally in focus under lowest power?
Can you specify the overlapping (superposition) order of the threads?
If yes, which thread sitson top and which one sits at the bottom?
3. Change to the low power objective (10x) and focus on the point of intersection. Are all three
threads equally in focus?
Can you specify the overlapping (superposition) order of the threads? If yes, which thread sits
on top, and which one sits at the bottom?
4. Change to the high- p o w e r objective (40x) and focus on two intersecting threads. Are
both threads equally in focus?
Can you specify the overlapping (superposition) order of the threads? If yes, which thread sits
on top, and which one sits at the bottom?
In order to see the individual threads clearly, is more focusing required under high or lowpower?

Experiment 3: Preparation of a wet mount
Purpose
To become familiar with the wet mount which consists in preparing a sample between a slide and a
cover slip with the addition of water between the sample and the cover slip.
Materials Required
- Slides and cover slips
- Onion epidermis
- Plastic Pasteur dropper
Procedure
One of the major advantages of the light microscope is that it permits the observation of
microscopic live tissues or organisms. For instance, the microscope enables us to observe
unicellular organisms swim about in the same way as we might watch a duck swim on a pond.
Living cells or any fresh material can be observed under the microscope by making a wet mount
on a slide.
1. Obtain a clean slide and cover slip.
2. Using forceps and a scalpel, remove a thin layer of epidermis from the inner side of an onion
slice and lay it flat on the center of the slide (beware with the way you use the scalpel).
3. Add two drops of water on top of the epidermis layer.
4. Hold the cover slip at an angle by two of its corners and touch one edge of the slide as in Fig.2.
5. Slowly pull the cover slip edge across the slide until it meets the water. The water will then
spread along the edge of the cover slip.
6. Carefully lower the cover slip over the slide using your fingers or a needle probe. The water
will spread evenly under the cover slip thus preventing any air bubble formation. If you have
any air bubbles under your cover slip, prepare a new wet mount.
Fig. 2. Wet mount preparation procedure.
7. Place the wet mount on your microscope stage and observe it with the lowest power objective
by using the focusing technique.

8. Observe the epidermis under low and high power. Adjust the iris diaphragm to a brightness that
reveals the most detail in the leaf. Note that the onion epidermis resembles a brick wall with
each “brick” consisting of a cell. The obvious boundary that outlines each cell is the cell wall
found in all plant cells, but which is absent in all animal cells.
9. When done, clean the wet mount slides and dispose of the cover slip in the yellow sharp box.

Experiment 3: Cell Staining

Experiment 3: Cell staining
Definition
Cell staining is a technique used to observe, analyze, and describe the structure and the
components of tissues using a microscope. By using different stains, one can stain certain cell
components, such as the nucleus or the cell wall, or the entire cell. Cells may also be stained to
highlight metabolic processes or to differentiate between live and dead cells in a sample. Most
stains are used on fixed (non-living) cells, while only some can be used on living cells.
Purpose of the staining technique

A tissue is stained and then observed under the microscope for the following reasons:
* It enhances the visualization of the sub-components of a cell or tissue.
* It accentuates specific morphological features.
* It differentiates between live and dead cells within a sample.
* It highlights metabolic processes.
Functions of the staining dyes

Histological staining involves the use of dyes to highlight specific intra- or extra-cellular
components within the tissue. A dye is a colored substance that has an affinity to the substrate
to which it is being applied. It is generally applied in an aqueous solution, and may require a
mordant to improve the fastness of the dye. These enhancing compounds are most often metal
salts. The mechanism of action of the mordants is not clear, but it presumably involves coordination
bonding between the metal and the dye, and then further coordination between this complex and the
tissue. A vast array of dyes and associated staining protocols are available for laboratory use. Each
dye is targeted toward different cellular structures.
The chemistry of staining: types of dyes

Staining procedures provide conditions which promote the binding of a given dye to specific
cellular or extracellular components. The utility of a staining procedure lies in its ability to bind
dye only to selected structures, highlighting these structures in contrast with the rest of the section
(the section is the piece of tissue applied onto the glass slide). To accomplish this, each procedure
makes use of a subset of possible interactions between the dye and the cellular components.
Ionic bonds are the predominant mode of interaction between tissues and dyes. They result from the
attraction between positive and negative charges. In solution, acidic groups [carboxylic (COO-); or
sulfonic acids (SO3-), or others] will lose a proton and become negatively charged (anionic). They
bind to basic tissue components. Eosin falls into this category of dyes and shows up in the basic
parts of the cell, i.e., the cytoplasm. Basic groups [generally amines (NH2+)] will accept a proton to
become positively charged cations. They bind to acidic tissue components such as nuclei.
The pH of the solution determines the extent to which any chemical group is protonated or
deprotonated, and a dye may have many such groups on its surface. Thus, altering the pH of a
staining solution will alter the charges on the dye and the tissue molecules, and therefore alter the
staining pattern. Crystal violet, methylene blue, and safranin fall into the basic dye category.
Neutral groups also exist. These groups are produced by the interaction of an acid and basic dye
producing a large molecule. These dyes are usually soluble in alcohol and possess some of the
staining properties of both dyes. E.g., Giemsa

Sources of dyes
Natural sources: The majority of natural dyes are from plant sources (berries, bark, leaves), wood,
fungi, and lichens. Plant-based dyes such as woad, indigo, saffron, and madder are also widely used
in cell staining.
Synthetic sources: Synthetic dyes quickly replaced the traditional natural dyes. They cost less, offer
a vast range of new colors, and impart better properties to the dyed materials.
Staining of a tissue mounted on a slide

There are 4 major steps involved in slide staining:
1- Permeabilization: sample is treated with a surfactant to dissolve cell walls and allow a
largeramount of dye molecules to penetrate the cell.
2- Fixation: it serves to "fix" or preserve cell or tissue morphology through the preparation
process. In this step, a chemical fixative is added to create chemical bonds between proteins and
enhance their rigidity. Common fixatives include formaldehyde, ethanol, methanol, and picric acid.
3- Mounting: it involves slicing then attaching samples onto a glass microscope slide for
observation.
4- Staining: it consists in the application of a stain to the tissue sample. This process may involve
immersing the sample (before or after fixation or mounting) in a dye solution and then rinsing and
observing the sample under a microscope.
5- To complete the mounting and protect the tissue, cover it with a cover slip.
Fig. 1: To mount a tissue, (A) Apply the mounting medium on top of the tissue section. (B) Hold the cover slip at 45°
thus allowing the drop to spread along its edge. (C) Lay slowly the cover slip on top of the tissue.
General types of staining

Simple stain: makes use of a single dye and reveals basic cell shapes and arrangements.
Differential stain: makes use of two or more dyes thus allowing the discrimination among
organisms based on structural differences.

Various applications of the staining technique
a- Histology
It is the study of the microscopic anatomy of cells and tissues in plants and animals. Knowledge of
histology shows how various cells are organized to form tissues and how each cell and tissue diplay
modifications according to their functional characteristics. Knowledge of histology is crucial to the
proper diagnosis of certain diseases like cancer.
Hematoxylin and eosin stain (H&E stain) is the most

commonly used staining in histology. Hematoxylin is a
natural compound extracted from a species of tree found in
Mexico and the West Indies. The extracted compound is
oxidized to produce hematein, which is the active staining
component of the hematoxylin stain. Therefore,
hematoxylin must be 'ripened' by oxidation before it can
be used. Hematoxylin staining requires the use of a
mordant (most commonly aluminum salts), targets the Fig. 2. H&E tissue stain.
cell nucleus, and produces a dark blue color.
Hematoxylin is used in combination with eosin because eosin stains the cytoplasmic organelles in
varying shades of pink, red or orange. The combination of the two stains will provide
morphological information about the tissue studied.
b-Hematology
It consists in the study of blood, blood-forming organs and blood diseases. Hematology includes
the study of etiology, diagnosis, treatment, prognosis, and prevention of blood diseases.
The May-Grünwald Giemsa is the most commonly used test in hematology and is the traditional
nuclear dye. It contains two dyes: methylene blue and eosin Y. Upon oxidation, methylene blue
produces a colored compound known as Azure that is blue violet and stains mainly acidic cell
components. Therefore, nuclei will be stained in varying shades of purple. In contrast eosin is pink
and stains basic structures.
Fig. 3. May-Grünwald Giemsa staining of a blood smear.
c- Cell culture
It is the complex process whereby cells are grown under
controlled conditions. In practice, the term "cell culture"
refers to the culturing of cells derived from eukaryotes.
However, there are also cultures of plants, fungi, and
microorganisms, including viruses, and protists. Cells are
grown under appropriate conditions (typically, 37°C, 5%
CO2 for mammalian cells) in a cell incubator. Culture
conditions vary widely from one cell type to another. Fig. 4. Principle of staining with Trypan blue.

The dye exclusion test is used to determine the number of viable cells present in a cell
suspension. It is based on the principle that live cells possess intact cell membranes that exclude
certain dyes, such as trypan blue, Eosin, or propidium. In contrast, non-viable cells will absorb the
dye and stain in blue. The most popular dye reagent used in the trypan blue. In this test, a cell
suspension is mixed with trypan blue and then visually examined to determine whether cells took
up or excluded the dye.
d- Bacteriology
It consists in the study of bacterial microorganisms. This sub-
category of microbiology involves the identification,
classification, and characterization of bacterial species.
The Gram staining technique is used to differentiate bacteria

into two main classes, gram-positive and gram-negative. A heat
fixed bacterial smear is stained with crystal violet (methyl
violet), treated with 3% iodine/potassium iodide solution,
washed with alcohol, and counterstained.
Fig. 5. Gram-stained bacteria.
e- Molecular Biology
It is the branch of biology that addresses the molecular basis of biological activity. This field
overlaps with other areas of biology and chemistry, particularly genetics and biochemistry.
Molecular biology mainly aims at understanding the interactions between the various systems of a
cell, including the interactions between the different types of DNA, RNA, and protein biosynthesis.
The most commonly used stain for detecting DNA/RNA is Ethidium Bromide (EtBr).
Ethidium Bromide is a DNA intercalating agent that inserts

itself between the base pairs of the double helix. EtBr
possesses a UV absorbance maxima at 300 and 360 nm and
re-emits this energy as a yellow/orange light centered at 590
nm. The major drawback of EtBr is that it is a potent
mutagen. Therefore, solutions must be handled with caution
and decontaminated prior to disposal.
f- Parasitology
It is the study of parasites, their hosts, and the relationship
between them.
Iodine is used to mainly stain the nuclei of cysts if present.
Cysts will absorb the iodine and appearlight yellow-gold in
color.
Fig. 6. Parasitic cyst stained with iodine.

Experiment 4: Miscellaneous Staining and Cell Diversity

A- Staining Techniques:
1- Staining of an Onion Epidermis Sample:
Purpose:
Plant cells carry a great variety of specialized structures that have various functions. These cellular
structures are generally referred to as organelles, or “little organs,” that are comparable to the
organs that keep the human body functioning. A great majority of organelles are colorless making
them essentially invisible under the microscope. Many kinds of special stains have been developed
that color specific cell organelles making them visible under the microscope. Some organelles, such
as the chloroplasts, are an exception because they are naturally pigmented. Indeed, they contain the
green pigment chlorophyll which gives plants their green color. In this experiment, students will
compare stained and unstained slices of onion epidermis to make a real-life observation of the
importance of staining techniques in the field of biology.
Materials Required
- Glass slides
- Glass cover slips
- Forceps
- Onion slices
- Dropper bottle of water
- Dropper bottle of acetocarmine stain
Procedure
1. Prepare a slide, a cover slip, and a small slice of onion.
2. Use forceps to carefully remove a thin layer of epidermis, or “skin”, from the inner curved side of
an onion slice (see Fig. 1.)
3. Place the thin epidermis on the slide and flatten it to avoid formation of any folds or wrinkles.
4. Make a wet mount of the onion epidermis: add a drop of water and then a cover slip on top of it.
5. Observe the onion epidermis under low and high power. Do you see cell walls in the epidermis?
Do you see chloroplasts? Why?
6. In parallel, prepare another slice of onion epidermis and add to it a drop of acetocarmine. Cover
with a cover slip and allow the stain to penetrate the onion skin for three minutes. Observe the
onion cells again under low and high power. The red acetocarmine stain is absorbed by the cell
nucleus making it appear as a small red circle within the cell.
7. Dispose of the slide and coverslip in the yellow sharp box
Fig. 1. Microscopic observation of an onion epidermis stained with acetocarmine.

2- The May-Grünwald-Giemsa Stain
Purpose
To become familiar with the May-Grünwald-Giemsa stain. This stain is used in the field of
hematology to perform tests on blood samples, one example is the counting of blood cells in a
blood sample or the monitoring of the shape of the cells. It combines two stains: the May-
Grünwald stain and the Giemsa stain. These are neutral mixtures with very distinct properties.
They are not active in an alcoholic medium and only act selectively when released in a buffered
aqueous solution. This releasing induces the precipitation of neutral stains. The May-Grünwald
stains acidophilic elements and the neutrophilic granulations of the leukocytes. The Giemsa
stains the cytoplasm of monocytes and lymphocytes as well as the chromatin of nuclei.
Procedure
1. Fix air-dried smear preparations for 5 minutes in a Coplin jar containing 100% Ethanol.
2. Air dry the slide 2 minutes at room temperature.
3. Stain 5 minutes in a Coplin jar containing undiluted May-Grünwald stain.
4. Wash three times, 2 minutes each, in a Coplin jar containing 1x phosphate-buffered water.
5. Stain 10 minutes in 5% Giemsa stain.
6. Rinse under cold running tap water.
7. Air dry the slide 10 minutes.
8. Examine the slide under l00x magnification power using oil immersion.
Staining characteristics
Nuclei Purple
Cytoplasm
Erythrocytes Deep pink
Reticulocytes Grey-blue
Neutrophils Orange-pink
Lymphocytes Blue; some small lymphocytes deep blue
Monocytes Grey-blue
Basophils Blue
Granules
Neutrophils Fine purple
Eosinophils Red-orange
Basophils Purple-black
Monocytes Fine reddish (azurophil)
Platelets Purple

Fig. 2. Microscopic observation of a blood smear staining by May-Grünwald Giemsa.
Fig. 3. Microscopic observation of a normal erythrocytes vs. sickle celled erythrocytes.
Fig. 4. Microscopic Observation of reticulocytes.

3- The Gram stain

Principle
Bacteria can be classified according to a specific hierarchy that ranges from the domain (least
specific) to the species (most specific). This classification depends on a number of criteria among
which the morphology (shape) of the bacteria analyzed. The most popular test to analyze the
morphology of bacterial cells is the Gram staining. This is the starting point for bacterial
identification procedures. In the late 1800’s, Christian Gram observed that some genera of bacteria
retained a dye-Iodine complex when rinsed with alcohol, while other genera were easily
decolorized with alcohol and could be visualized only after a contrasting counterstain. This
staining procedure defines two bacterial groups: those which retain the primary dyes (“Positive by
Gram’s Method” or “Gram-Positive”) and those which are easily decolorized (“Negative by
Gram’s Method” or “Gram-Negative”).
The difference in dye retention is dependent on physical properties such as thickness, density,
porosity, and integrity of the bacterial cell wall, as well as, to some extent, the chemical
composition:
- Gram-Positive bacteria have thick, dense, relatively non-porous walls, while Gram-Negative
bacteria have thin walls surrounded by lipid-rich membranes.
- Some non-bacterial organisms with thick cell walls (e.g., some yeasts) also stain Gram-Positive.
- Gram-Positive bacteria which have lost wall integrity through aging, or physical or chemical
damage may stain Gram-Negative.
Procedure
Step 1: Prepare a Smear
- suspend some of the material to be stained in a drop of water on a microscope slide.
- spread the drop to about the size of a nickel.
- allow to air dry.
- heat fix by gently warming above a flame or another source of heat.
Step 2: Apply the Primary Stain
- cover the smear with Crystal Violet.
- allow standing for 30 seconds.
- rinse with water to remove excess stain.
Step 3: Apply the Fixing Agent
- cover the smear with Iodine solution.
- allow standing for 1 minute.
Step 4: Rinse
- rinse with water to remove excess Iodine.
Step 5: Decolorize
- drip 95% Alcohol across the slide for 5 seconds exactly.
- the effluent should appear pale or clear.
Step 6: Rinse
- rinse with water to remove the excess of alcohol.
Step 7: Counterstain
- flood the slide with Safranin solution.
- let stand for 30 seconds.

Step 8: Rinse, dry, and observe.
- rinse with water to remove excess stain.
- blot dry.
- observe under 100x magnification using oil immersion.

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BIO210L

Cells and Molecules Laboratory

Introduction: Guidelines for Writing a Lab Report ................................................................................................ 3

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B- The content of the lab report

B.1. Cover page

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(9 lines, Line spacing = “Single”)

(4 lines, Line spacing = “Single”)

(22 lines, Line spacing = “Single”)

Name: Your Name

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C- Template for a lab report

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Time (min.) Group 1 (+B.S.1) Group 2 (+Pancreatin) Group 3 (+B.S.+ Pancreatin)

* mean pH of 3 sample tubes per group

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A- General Biology Lab Safety Rules:

1. Open-toed shoes are not permitted.

B- While Working in the Lab:

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- Never carry more than one container in one hand.

D- Safety Procedures Related to the Manipulation of Equipment:

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E- Biohazard and Chemical Waste Disposal:

1. Always dispose of chemical waste in the allocated storage containers.

F- In Case of Accidents in the Lab:

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B- Major types of microscopes

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Fig.1. Anatomy of a compound light microscope.

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Fig.5. SEM of blood cancer cells.

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B- Practical use of the microscope

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Experiment 1: Adjusting the focus and determining the image orientation.

5. Look at the magnified image of the letter.

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Experiment 2: The concept of depth of field.

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Experiment 3: Preparation of a wet mount

Fig. 2. Wet mount preparation procedure.

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Purpose of the staining technique

Functions of the staining dyes

The chemistry of staining: types of dyes

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Staining of a tissue mounted on a slide

General types of staining

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