BIOLOGY I

LABORATORY MANUAL

Name: ______________________________________________

Group: ______________________

MSc Cecilia Y. Garibay Flores

July 2021
 Biology I Lab Manual

Content

Introduction 2

Lab Course Materials 2

General Rules 3

Lab and Field Safety Rules 4

Lab Grading Summary 5

Lab Grading Rubric 6

Lab Practices Calendar 7

1. The microscope as a Tool in the Biology Lab 8

2. Biomolecules: Carbohydrates 13

3. Biomolecules: Proteins and Lipids 16

4. Biomolecules: DNA 19

5. Cell Types: Prokaryotic and Eukaryotic Cells 21

6. Cell Transport: Osmosis 24

7. Mechanisms of energy generation: Photosynthesis 26

8. Mechanisms of energy generation: Cellular Respiration 31

Bibliography 35

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 Biology I Lab Manual
Introduction

The Biology I course aims to address topics of interest to students and form in them a biological
culture that allows them to reflect and participate actively and consciously in taking decisions in the
personal and social sphere, in a way that favors the preservation of life as a fundamental value of the
human being.

The Biology I lab practices are intended to integrate the theoretical knowledge acquired in the
classrooms with the laboratory; a place where the students develop their capacities on their own. The
laboratory is the testing site for the new ideas and hypotheses that advance science and the student
will be there to actively learn, reaffirm their knowledge and generate new learning.

Lab Course Materials

● Cotton Lab Coat

● Laboratory Manual (printed): It is each student’s responsibility to print out the entire document
– black and white, double-sided is fine.
● Specific materials will be asked for each lab practice in advance
● PreLab Work that includes a proposed hypothesis.

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 Biology I Lab Manual
General Rules

1. Come to the lab! Three unexcused absences and you fail the lab of the Biosciences course.
You cannot pass the Bioscience course if you fail the laboratory portion due to absence, so
make it a priority to attend the lab! If you know you are going to miss a lab, notify your teacher
immediately. Missed labs should be made up within one week. Documentation for your
absence must be provided to make up a lab and complete any assignments. While you can
make up labs outside of normal class time, quizzes may only be taken in a regularly scheduled
lab.

2. Read the lab manual before the session and bring your printed copy. Reading the lab manual
will provide insight into what you are doing in the lab. It gives you essential background
information and describes the protocols you will follow to complete lab experiments. If you read
the lab manual before coming to class, you will finish the lab faster and do better on
assessments. In addition, there will be a prelab work that is a requirement to do the laboratory
practice. There will be questions from the lab manual on each week’s lab quiz. You should take
notes on your manual

3. Watch the weekly video that links the lab to practice. These videos are intended to give each
lab context and meaning. They are examples of how you will encounter biology outside of the
classroom. The videos will be the basis for most of the content on your lab quizzes, so make
sure you give yourself adequate time to read each week. We encourage you to take notes on
these videos. Watching the video and reading the lab manual should take about 45
minutes/week.

4. Turn in your assignments through Classroom by the time the lab begins. All assignments must
be submitted through Classroom, email submission is not acceptable. In-class assignments
(exit passes) are due at the end of your lab section (or when you make up the lab) and cannot
be turned in later. Absence from the lab or scheduled travel is not an excuse for late
assignments.

5. Do your own work! Even though you will be working with others in the lab, every assignment in
this class (including exit passes) is your own work. Plagiarism will not be tolerated as stated in
the Harkness Institute Academic Rules. If there is evidence that any part of an assignment is
plagiarized, the work will be sent to the Ethics Committee for review. If you are found in
violation of Harkness Institute Academic Rules, you will receive a failing grade for the entire
Introduction to Biosciences course.

Laboratory and Field Safety Rules

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 Biology I Lab Manual
1. Conduct yourself in a responsible manner at all times.

2. Lab and safety information and procedures must be read ahead of time. All verbal and written
instructions shall be followed in carrying out the activity or investigation.

3. Eating, drinking, gum chewing, applying cosmetics, manipulating contact lenses, and other
unsafe activities are not permitted in the laboratory.

4. Working in the laboratory without the instructor present is prohibited.

5. Unauthorized activities or investigations are prohibited. Unsupervised work is not permitted.

6. Removing chemicals or equipment from the classroom or laboratory without instructor

permission is prohibited.

7. If you do not understand how or why to do a task, ask your instructor for help. If there is any
doubt in your mind, ask your teacher.

8. Dress appropriately for laboratory work by protecting your body with clothing and shoes. This
means that you should use hair ties to tie back long hair and tuck into the collar. Do not wear
loose or baggy clothing or dangling jewelry on laboratory days.Sandals or open-toed shoes are
not to be worn during any lab activities.

9. Know the location of all safety equipment in the room. This includes eye wash stations, the
deluge/safety shower, fire extinguishers. Know how to USE all safety equipment in the room,
and know the location of the Emergency Evacuation Route map.

10. Wash your hands with soap and water after handling any chemicals, glassware or touching any
surface in the lab area before leaving the lab area.

11. When an activity or investigation requires the use of laboratory gloves for hand protection, the
gloves shall be appropriate for the hazard and worn throughout the activity.

12. Avoid inhaling fumes that may be generated during an activity or investigation.

13. Never fill pipettes by mouth suction. Always use the suction bulbs or pumps.

14. Do not force glass tubing into rubber stoppers. Use glycerin as a lubricant and hold the tubing
with a towel as you ease the glass into the stopper.

15. Proper procedures provided by the instructor shall be followed when using any heating or flame
producing device. Never leave a flame unattended.

Laboratory Grading Summary

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 Biology I Lab Manual

Your lab grade is 20% of your Introduction to Biosciences course grade.

The criteria that will be evaluated in this Lab are the following:

Criteria Description Weight

PreLab Work A mandatory requirement to do the lab practice 15%

Exit Pass A summary written individually or an exit quiz 25%

Lab Report One per team 50%

One per Lab practice
See the Lab Report Rubric

Lab Performance Attendance, punctuality, appropriate dress, 10%

attitude, etc.

TOTAL 100%

LABORATORY REPORT RUBRIC

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 Biology I Lab Manual
Not
Excellent (4 pts) Good (3 pts) Adequate (2 pts) Needs Work (1 pt) attempted
(0)
1. Includes the title of the lab practice. One of the "excellent" Two of the "excellent" Introduction present,
2. Date of the la practice and submission conditions is not met, conditions is not met, no exemplary
Title and ID
date two conditions met one is met conditions met
data
3. Names of the members of the team
4. Course/class
1.Includes a short explanation of the One of the "excellent" Two of the "excellent" Introduction present,
theoretical basis or principle that was conditions is not met, conditions is not met, no exemplary
demonstrated in the experiment. two conditions met one is met conditions met
2. Includes the general procedure and if
Introduction
something was changed.
3. Includes an experimental question.
4. It is properly cited
5. 150-200 words
1. Includes a list of the lab materials and Description included, The description gives Would be difficult to
equipment. some steps are vague generalities, enough for repeat, reader must
2. Includes a list of reagents, the safety or unclear reader to understand guess at how the
Material and
measures to handle and dispose them how the experiment was data was gathered or
Methods
3. Description or flow chart of the conducted experiment
process is included (It could be repeated conducted
by another scientist)
1. Results and data are clearly recorded Results are clear and Results are unclear, Results are
and organized. labeled, trends are not missing labels, trends are disorganized or
2. Includes pictures and tables that obvious or there are not obvious, poorly recorded, do
Data and
summarize the results, so it is easy for minor errors in disorganized, there is not make sense ; not
Analysis
the reader to see trends. organization enough data to show the enough data was
3. All appropriate labels or descriptions experiment was taken to justify
are included. conducted results
1. Compare and contrast your results
with the theory.
2. Justifies if the hypothesis was
Discussion
accepted or not.
3.Discusses applications or real-world
connections.
1. Summarizes data used to draw 3 of 4 of the "excellent" 2 of the 4 excellent 1 of the 4 excellent
conclusions conditions is met conditions met conditions met
2. Conclusions follow data (not wild
Conclusions
guesses or leaps of logic),
3. Hypothesis is rejected or accepted
based on the data.
Student did not
Some of the conditions
follow directions,
Lab report submitted as directed, and on Most of the excellent met, directions were not
Format and practiced unsafe
time. Directions were followed, stations conditions were met; explicitly followed, lab
Lab procedures, goofed
were cleaned. All safety protocols possible minor errors in stations may have been
Protocols around in the lab, left
followed. format or procedures left unclean or group not
a mess or equipment
practicing good safety.
lost
3 bibliographies in APA
Bibliography 4 bibliographies in APA style 2 bibliographies No bibliography
style
No grammar or spelling mistakes.
WOW No typing mistakes.
points: Use the same font type and size)
-1/100 penalty for each mistake.

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 Biology I Lab Manual

Lab Practices Calendar

Number Lab Practice PreLab Work Exit Pass Week

1 The Microscope as a Tool in Yes Yes

the Biology Lab

2 Biomolecules: Carbohydrates Yes Yes

3. Biomolecules: Yes Yes

Proteins and Lipids

4. Biomolecules: DNA Yes Yes

5. Cell Types: Prokaryotic and Yes Yes

Eukaryotic Cells

6. Cell Transport: Osmosis Yes Yes

7. Mechanisms of energy Yes Yes

generation: Photosynthesis

8. Mechanisms of energy Yes Yes

generation: Cellular
Respiration

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 Biology I Lab Manual
1. The Microscope as a Tool in the Biology Lab

Team:

Introduction
The microscope is the most important instrument for biologists. It enables scientists to investigate
worlds not visible to the human eye. This is an opportunity to learn what it is and how to use the
microscope. The resolution power of microscopes increases up to 4000 times the actual size of the
observed object.
Thanks to the microscope, countless organisms have been studied, such as the cell. Its contribution to
Science has been and is of incalculable value and its development in these last three centuries has
allowed
expand the field of research, becoming the basic instrument to open borders in the field of Biology.

The parts that form in a compound microscope are grouped into three systems:
a) Mechanical System: It comprises the following parts: foot, column, arm, microscope tube, coarse
screw, micrometer screw, tweezers or clips and stage.

b) Optical System: It comprises two lens systems: the objectives and the eyepiece.

c) Lighting System: It serves to give, reflect, condense and control the amount of light that will pass
through the specimen to be observed and is placed under the stage. Understand the lamp, condenser
and diaphragm or iris.

Objectives:
Students will be able to:
● Identify the parts and the care that must be taken when using the microscope.
● Distinguish through experimental activity, the functioning of each of the microscope
components.
● To do simple preparations for microscopy

PreLab Work
Watch the following video: https://youtu.be/SUo2fHZaZCU
Draw a flow sheet of the use of a microscope.

Hypothesis:

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Materials and Equipment
1 Microscope
3 Coverslips
3 Slides
3 Samples to observe

Procedure
1. Identify the microscope parts:

Objective Coarse focus Stage Stage clips Base

knob

Eyepiece lens Arm Fine focus knob Light source Diaphragm

(ocular)

2. Preparing a wet mount of the letter "e”.

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 Biology I Lab Manual
● With your scissors cut out the letter "e" from the newspaper.
● Place it on the glass slide
● Cover it with a clean cover slip.
● Using your eyedropper, place a drop of water on the edge of the cover slip where it
touches the glass slide. The water should be sucked under the slide if done properly.
● Turn on the microscope and place the slide on the stage; making sure the "e" is facing
the normal reading position (see the figure above). Using the coarse focus and low
power, move the body tube down until the "e" can be seen clearly.
● Describe the relationship between what you see through the eyepiece and what you
see on the stage
● Calculate the total magnification.

3. Preparing a temporary mount of an onion peel

● Separate the epidermis (transparent layer) to a piece of onion, place it in a slide by
adding a drop of water and then placing a coverslip, make sure there are no air bubbles
or excess liquid.
● Place the preparation on the microscope stage, taking care to center the object at
observation, then fix it with the platen clamps to prevent the slide.
● Place the lowest magnification target in a vertical position, bring it as close as possible
to the
● preparation by means of the coarse screw, check this operation laterally, never looking
through the eyepiece.
● Look through the eyepiece as close to the front lens as possible, raising the tube of the
microscope with the coarse screw until an image is obtained; this is done with a smooth
movement so that you focus the image well.
● Once the image is located, if it lacks precision, turn the fine focus coarse until you get
the right focus.
● Immediately turn the revolver to change the objective for the one with the highest
magnification, taking care that it is well centered in the notch.
● Describe the relationship between what you see through the eyepiece and what you
see on the stage
● Calculate the total magnification.

Observations and Results

Sketch your observations using at least 2 different objectives and calculate the total magnification.

Drawing is a very important skill in biology and is considered a type of data collection because
drawings help to record data from specimens.
Drawings can highlight the important features of a specimen. A drawing is the result of a long period of
observation at different depths of focus and at different magnifications

Drawing Materials: All drawings should be done with a sharp pencil line on white, unlined paper.
Diagrams in pen are unacceptable because they cannot be corrected.

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 Biology I Lab Manual

Use a ruler to draw straight, horizontal lines. The labels should form a vertical list.
Technique: Lines are clear and not smudged. Avoid ‘feathery’ pencil lines and gaps. There are almost
no erasures or stray marks on the paper. Color is used carefully to enhance the drawing.

Accuracy: Draw what is seen; not what should be there. Avoid making “idealized”drawings. Do not
necessarily draw everything that is seen in the field of view. Draw only what is asked for. Show only as
much as necessary for an understanding of the structure – a small section shown in detail will often
suffice. It is time consuming and unnecessary, for example, to reproduce accurately the entire
contents of a microscopic field.

Title: The title should state what has been drawn and what lens power it was drawn under (for
example, phrased as: drawn as seen through 400X magnification)

Scale: Include how many times larger the drawing is compared to life size and a scale line that
indicates relative size.

Letter “e”

______________

________________

Eyepiece magnification X Objective magnification = Total magnification

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 Biology I Lab Manual

Onion peel

______________

________________

Eyepiece magnification X Objective magnification = Total magnification

Conclusions
Write your conclusions based on your results, the objectives of the lab practice and if your hypothesis
was accepted or not.

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 Biology I Lab Manual

2. Biomolecules: Carbohydrates

Team:

Introduction
There are four important types of large carbon-based molecules in living organisms- proteins,
carbohydrates (sugars & starches), lipids (fats), and nucleic acids.
Proteins, carbohydrates, and fats serve as nutrients in the food that we eat. Carbohydrates are used
by living organisms as an important source of energy. Simple carbohydrates are made of carbon,
hydrogen, and oxygen atoms in a 1:2:1 ratio. This ratio means that for every one carbon atom present
in the carbohydrate, there are two hydrogen atoms and one oxygen atom present. The monomers of
carbohydrates are referred to as monosaccharides.
Common examples of monosaccharides include glucose, fructose, and lactose. Sucrose, or table
sugar, and lactose, the sugar found in milk, are double sugars made from two monosaccharides.
Benedict’s Solution can be used to test for the presence of a monosaccharide, like glucose. In the
presence of glucose, Benedict’s Solution turns from blue to orange when heated. When many
monosaccharides join together, the resulting molecule is called a polysaccharide. Important
polysaccharides include cellulose, starch, and chitin.
Lugol’s Solution or Iodine can be used to test for the presence of a polysaccharide, like starch.In the
presence of starch, Lugol’s Solution turns from yellowish brown to purple or black.

Objectives
The student should be able to:
Identify the functional groups for each of the biomolecules that react in the following biochemical tests:
Benedict’s test and Iodine test.
Describe the mechanism of reaction for the Benedict’s test and the Iodine test.
Interpret the results when presented with data for each of the biochemical tests.

PreLab Work
Investigate the structure of five monosaccharides classifying them as hexoses or pentoses.
Investigate the structure of four polysaccharides indicating their monomeric units
Investigate what is the composition of the Fehling reagent, the Benedict reagent and the Lugol

Hypothesis:

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 Biology I Lab Manual

Material and equipment

4 Test tubes
Porcelain plaque with 6 wells
Glucose solution
Sucrose solution
Starch solution
Unknown carbohydrate sample
Alcohol lamp
Water Bath at 80°C
Benedict or Fehling Reagent
Iodine solution (Lugol)
Stirring rod
Thermometer
2 Pasteur pipettes

Procedure
Monosaccharide test
1. Label 4 test tubes as follows:
Test tube #1: distilled water
Test tube #2: glucose solution
Test tube #3: sucrose solution
Test tube #4: starch solution

2. Add 1ml of the Benedict or Fehling Reagent solution to each test tube.
3. Record your observations in the results table
4. Place all the test tubes in the water bath at 80°C (be careful not to get burned) during 5 min
5. Record your observations in a table in the results section.

Polysaccharide test

1. Label the porcelain plate as follows

Well #1: distilled water
Well #2: glucose solution
Well #3: sucrose solution
Well #4: starch solution
2. Add 2 drops of the Iodine’s reagent to each sample.
3. Record your observations in a table in the results section

Unknown carbohydrate sample

1. Record the physical appearance of your sample
2. Do the simple carbohydrate test
3. Do the polysaccharide test.
4. Record your observations and identify the type of carbohydrate is present in your sample.

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 Biology I Lab Manual

Observations
Sketch and describe your observations.

Results

Sample w/Benedict (+ /-) room w/Benedict (+/-) at Iodine solution

temperature 80°C

Distilled water

Glucose solution

Sucrose solution

Starch solution

Unknown carbohydrate

Conclusions
Write your conclusions based on your results, the objectives of the lab practice and if your hypothesis
was accepted or not.

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 Biology I Lab Manual
3. Biomolecules: Lipids and Proteins

Team:

Introduction
Lipids are also made of carbon, hydrogen, and oxygen but the ratio of carbon, hydrogen, and oxygen
atoms is not 1:2:1. Instead, lipids have a much greater number of carbon and hydrogen atoms with few
oxygen atoms present. their individual subunits (monomers) are fatty acids and glycerol. Lipids are
organic compounds that are not soluble in water, examples include fats, oils, and the wax covering
leaves.

The nonpolar bonds that form between the carbon and hydrogen atoms of a lipid cause them to be
hydrophobic or “water repellent” molecules, as opposed to hydrophilic or “water loving” molecules.
This attribute explains why water and oil do not mix. Sudan III Solution can be used to test for the
presence of a lipid. In the presence of a lipid-rich solution and water, Sudan III Solution forms a distinct
layer or clump in the well or test tube (layers or clumps is positive, no layers or clumps is negative).

The large number of carbon to hydrogen bonds also serves to make lipids energy-rich storage
molecules. One gram of a lipid stores twice as much energy as one gram of carbohydrate or protein.
Lipids from animals are referred to as fats and are solid at room temperature, whereas those found in
plants are referred to as oils and are liquid at room temperature. Fats and oils are triglycerides, which
are biomolecules that are composed of glycerol and three fatty acids.

One important relative of triglycerides are phospholipids. Phospholipids differ in structure from regular
triglycerides in that phospholipids are made of a glycerol and two fatty acids. A charged phosphate
group replaces the third fatty acid. The arrangement causes phospholipid molecules to have both
hydrophilic and hydrophobic regions. This feature also makes phospholipids an ideal structural
component of cell membranes.

Proteins are made of monomers called amino acids, which are composed of atoms of carbon,
hydrogen, oxygen, and nitrogen. Proteins are large and complex molecules that combine to form
various components of living organisms such as muscle fibers, enzymes, and hemoglobin. Proteins
are made from specific sequences of amino acids. A string of amino acid monomers joined together by
peptide bonds is called a polypeptide. Biuret’s Solution (Cu2SO4 + NaOH) can be used to test for the
presence of protein. In the presence of protein, Biuret’s Solution turns from blue to violet (violet is
positive, blue is negative).

Objectives
Determine qualitatively proteins and lipids.
Establish experimentally some chemical and physical properties of proteins and lipids

Materials and Equipment

Distilled water
Crystals of Sudan III
Cooking oil
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 Biology I Lab Manual
Mayonnaise
10% egg yolk suspension
10% egg albumin
10% milk
Porcelain plaque with 6 wells
5 test tubes
5 mL pipettes
pears for pipettes
droppers
Spatula
Brown paper bag

PreLab Work
Investigate the physical properties of lipids and proteins

Hypothesis

Procedure
Lipids test

1. Draw 6 circles spaced 5cm apart on a piece of brown paper, paper bag, or brown envelope.

2. Place a drop of each sample on the brown paper or use another small piece of bag to crush the
solid samples into the circle to release its liquid content.

3. Allow paper to dry for 15 min, then turn over the paper to the opposite side to review your
results. Do you see a stain (oil) for the below samples?

4. Create a chart and record your observations.

Sudan III Test for Lipids

1. Place 10 drops of each sample in each well of the porcelain plaque

2. Add 5 drops of Sudan III Solution to each well.

3. Observe and record the results in the Data Table.

Protein Test

1. Label the test tubes with the name of each sample.

2. Place 1ml of each sample in the proper test tube.

3. Slowly add about 10 drops of the Biuret’s solution to each test tube and mix carefully.

4. Observe and record the results in the Data Table.

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 Biology I Lab Manual
Observations
Sketch your experiments and describe them

Results

Sample Brown Paper Test (+/-) Sudan III Test Biuret Test

Distilled water

Cooking Oil

Mayonnaise

Egg Yolk

Egg Albumin

Milk

Conclusions
Write your conclusions based on your results, the objectives of the lab practice and if your hypothesis
was accepted or not.

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 Biology I Lab Manual

4. Biomolecules: DNA

Team:

Introduction
All living organisms have DNA, which is short for deoxyribonucleic acid; it is basically the blueprint for
everything that happens inside an organism’s cells. Overall, DNA tells an organism how to develop
and function, and is so important that this complex compound is found in virtually every one of its
cells.Each cell has an entire copy of the same set of instructions, and this set is called the genome.
Scientists study DNA for many reasons: They can figure out how the instructions stored in DNA help
your body to function properly. They can use DNA to make new medicines or genetically modify crops
to be resistant to insects.
During a DNA extraction, a detergent will cause the cell to pop open, or lyse, so that the DNA is
released into solution. Then alcohol added to the solution causes the DNA to precipitate out. In this
activity, strawberries will be used because each strawberry cell has eight copies of the genome, giving
them a lot of DNA per cell.

Objective
Students will be able to extract DNA and test some of its properties.

PreLab Work
Investigate the chemical structure of DNA and the organization within the cell

Hypothesis

Material and Equipment

Rubbing alcohol
Measuring cup
Measuring spoons
Salt
Water
Dishwashing liquid (for hand-washing dishes)
Glass or small bowl
Cheesecloth or filter paper
Funnel
Beaker
Three strawberries
Resealable plastic sandwich bags

Procedure

1. Before getting started, put a 1/4 cup of rubbing alcohol into the freezer.

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 Biology I Lab Manual

2. Cut up your fruit into tiny piece and place them inside the plastic bag

3. Add a tablespoon of dish soap and a teaspoon of salt.

4. Add 1/3 of a cup of water.

5. Seal up your bag and squish all the ingredients together until it makes a pulp.

6. Place a paper filter over the top of your funnel and place it over the beaker.

7. Pour the filtered juice into your test tube.

8. Cover the juice with a 5ml of rubbing alcohol.

9. Take the DNA with a Pasteur pipette and place a sample in a different test tube.

10. Observe it under a microscope.

Observations
Sketch and describe your observations.

Conclusions
Write your conclusions based on your results, the objectives of the lab practice and if your hypothesis
was accepted or not.

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6. Cell Types: Prokaryotic and Eukaryotic cells

Team:

Introduction
Living beings are made up of cells; some multicellular, such as plants and animals, but others like
bacteria, are unicellular and therefore invisible, so they could not be observed until the introduction of
the microscope. There are two kinds of cells: prokaryotes (bacteria) and eukaryotes.
Prokaryotic cells (bacteria) are microorganisms which are not easy to distinguish when microscopy,
this is because they have little contrast, hence the need to stain them in order to increase the contrast
with your surroundings.
Eukaryotic cells can be easily observed under the microscope, just make a thin section of the
specimen and prepare to be able to observe it.

Objectives
The student will be able to observe in the microscope, both prokaryotic cells as eukaryotic cells. And in
turn determine the morphological differences fundamental between the two.

PreLab Work
Write a summary of the characteristics of each type of cell

Hypothesis

Material and Equipment

Fresh Onion
Yoghurt
Dissection forceps
Slides
Cotton
Distilled water
Alcohol lamp
Immersion Oil
Clean lens solution
Violet dye
Lugol
Microscope
Unknown sample

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Procedure

Prokaryotic cell Observation

1. Label one clean slide

2. Place a drop of yoghurt

3. Place a drop of water and carefully mix it

4. Carefully fix the prepared slide in the flame of the alcohol lamp (5 seconds max)

5. Cover the preparation with 5 drops of the violet dye during 1 minute.

6. Rinse carefully with water.

7. Let air dry

8. Observe the preparation with the microscope

Eukaryotic cells
1. Label the slides.

2. Place the sample to be observed in the central part of the slide:

a) Plant Sample: Make a thin section of the sample and cover it with a drop of water.

3. Place the coverslip on top of the slide.

4. Observe under the microscope

Unknown sample:

1. Process the sample to identify the type of cell.

Observations

Draw the observations for each type of cell that you observe. Describe its morphology and describe
the major structures that you were able to observe.

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Conclusions
Write your conclusions based on your results, the objectives of the lab practice and if your hypothesis
was accepted or not.

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7. Cell Transport: Osmosis

Team:

Introduction
The plasma membrane has the important function of regulating the entry and exit of substances inside
and outside the cytoplasm as it has a normal composition that needs to be kept constant.
The lipid bilayer of the membrane acts as a barrier that separates two aqueous media, the medium
where the cell lives and the internal cellular environment. Cells require nutrients from the outside and
must eliminate waste substances from metabolism and thus keep your metabolism stable internal
environment. Its permeability is selective, since it allows the passage of small molecules and blocks
others.
Osmosis is a type of simple diffusion, where the water molecules follow their gradient of concentration
and move from an area where the concentration is high to an area where the concentration is low.

Objectives
● Understand the structure and the different transport mechanisms across the membrane mobile
● Understand the osmosis process
● Experiment with osmosis in solutions and identify which are isotonic, hypotonic, and hypertonic
and the effects of plasmolysis and turgor

PreLab Work

Hypothesis

Material and Equipment

Onion
Plant Leaves
Scalpel
Petri dishes
Pasteur pipettes
Slides
Coverslips
Distilled water
Mineral water
Isotonic solution
Optical microscope

Procedure

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1. Prepare 20 mL of a saturated NaCl solution.

2. Add 10 mL of saturated NaCl solution in a Petri dish, also add 10 mL of mineral water in a
different box and in another Petri dish add 10 mL of distilled water.

3. Label the boxes correctly.

4. With the scalpel cut a fragment (of the same size) of onion epidermis and do the same with the
leaf in each of the Petri dishes and allow 30 minutes to elapse. It is important that the solutions
fully cover the samples.

5. Remove the samples and place them on different slides with a little of the solution they were
placed in.

6. Look under the microscope at 10X and 40X.

Observations and Results

1. Make two representative diagrams of cells in Isotonic, Hypertonic and Hypotonic and describe
the observation in each.

2. Determine in what type of experienced solutions the phenomenon of plasmolysis was

observed, turgor or balance.

3. Explain how you can classify the type of solution (when and why water enters or leaves the
cells)

4. Make an outline of what you observed and explain.

Conclusions
Write your conclusions based on your results, the objectives of the lab practice and if your hypothesis
was accepted or not.

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8. Mechanisms of Energy Generation: Photosynthesis

Team:

Introduction
Plants transform sunlight to chemical energy by means of photosynthesis (photo=light,
synthesis=putting together). During the process, plants fix carbon dioxide from the atmosphere and
release oxygen and water (called transpiration). Measurements of photosynthesis are needed for
comparing and understanding productivity of vegetation and their response to environmental stresses.
Photosynthesis, generally, is the synthesis of sugar from light, carbon dioxide and water, with oxygen
as a waste product. It is the most important biochemical pathway known; nearly all life depends on it. It
is a complex process, consisting of many coordinated biochemical reactions. It occurs in most plants,
as well as algae, some bacteria, and some protists, organisms collectively referred to as
photoautotrophs.
Photosynthesis occurs in two stages. In the first phase light-dependent reactions (also called the Light
reactions) capture the energy of light and use it to make high-energy molecules. During the second
phase, the light-independent reactions (also called the Calvin-Benson Cycle, and formerly known as
the Dark Reactions) use the high-energy molecules to capture carbon dioxide (CO2) and make the
precursors of glucose.
In the light-dependent reactions the pigment chlorophyll absorbs light and loses an electron that
travels down an electron transport chain producing the high energy molecules NADPH and ATP. The
chlorophyll molecule regains its electron by taking one from a water molecule through a process called
photolysis that releases oxygen gas as a byproduct.
Photosynthesis may simply be defined as the conversion of light energy into chemical energy by living
organisms. It is affected by its surroundings and the rate of photosynthesis is affected by the
concentration of carbon dioxide, light intensity and the temperature.
There are three main factors that affect photosynthesis. These factors are: Light irradiance and
wavelength, carbon dioxide concentration, and temperature.

Objectives
In this experiment students:
● Will learn how to measure the rate of photosynthesis indirectly using the floating leaf disk
method.
● Will also investigate various factors that can affect the process of photosynthesis.

PreLab Work
1. What do plants need to live? What do they need to carry out photosynthesis (to
photosynthesize)?

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2. Fill in the table below with as many ideas as you can. The purpose of this activity is to find out
what you know before studying photosynthesis, so don’t worry about being right or wrong.

What Plants Need

What do plants need/use/consume? What do plants give off? (products)

(reactants)

3. Based on what you wrote above, write a summary equation for photosynthesis.

Hypothesis

Material and Equipment

Sodium bicarbonate
Liquid soap
Plastic syringes
Beakers or plastic cups
Transfer pipettes

Procedure

1. Prepare a 0.2% solution of sodium bicarbonate. (1/8 of a teaspoon of baking soda in 300 ml of
water)

2. Add 1 drop of dilute liquid soap to this solution. The soap wets the hydrophobic surface of the
leaf, allowing the solution to be drawn into the leaf. It’s difficult to quantify this since liquid
soaps vary in concentration. Avoid suds. If your solution generates suds, then dilute it with
more bicarbonate solution.

3. Cut 10 or more uniform leaf disks for each trial (single hole punches work well for this but stout
plastic straws will work as well).

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4. Infiltrate the leaf disks with sodium bicarbonate solution.

5. Remove the piston or plunger and place the leaf disks into the syringe barrel. Replace the
plunger being careful not to crush the leaf disks. Push on the plunger until only a small volume
of air and leaf disk remain in the barrel (< 10%).

6. Pull a small volume of sodium bicarbonate solution into the syringe. Tap the syringe to
suspend the leaf disks in the solution. They will all float.

7. Holding a finger over the syringe-opening, draw back on the plunger to create a vacuum. Hold
this vacuum for about 10 seconds. While holding the vacuum, swirl the leaf disks to suspend
them in the solution. Let off the vacuum. The bicarbonate solution will infiltrate the air spaces
in the leaf causing the disks to sink. You will probably have to repeat this procedure 2-3 times
in order to get the disks to sink. If you have difficulty getting your disks to sink after about 3
evacuations, it is usually because there is not enough soap in the solution. Add a few more
drops of soap.

8. Pour the disks and solution into a clear plastic cup. Add bicarbonate solution to a depth of
about 3 centimeters. Use the same depth for each trial. Shallower depths work just as well.

9. For a control, infiltrate leaf disks with a solution of only water with a drop of soap- no
bicarbonate.

10. Place under the light source and start the timer. At the end of each minute, record the number
of floating disks. Then swirl the disks to dislodge any that are stuck against the sides of the
cups. Continue for 15 minutes or when most of the disks are floating.

11. After all (or most) of the disks are floating, put the cups in the dark (a shoebox or a closet) and
monitor for the next 15 minutes.

12. Record how many disks remain floating after each minute until all (or most) of them have sunk.

Observations
Sketch your observations and describe them.

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Results
1. Record in the following tables how many disks are still floating at the end every minute for 15
minutes for the control conditions and with CO2

Control conditions With CO2

Minute Number of floating Number of floating Number of floating Number of floating

discs w/Light discs without Light discs w/Light discs without Light

10

11

12

13

14

15

2. Plot your results in a graph (independent variable in the “x” axis and dependent variable in the
“y” axis.

3. Answer the following questions:

● How does the suction help the leaf disks to sink?

● How does the detergent help the leaf disks to sink?

● Why don't the leaf disks soaking in the water (control) float?
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● What is the purpose of the baking soda solution?

● What is the purpose of the light reaction?

● Why do the leaf disks in the baking soda solution (treatment) begin to float?

● Why do the leaves begin to sink again in the dark?

● Why don't the leaves in the baking soda solution continue to produce oxygen in the dark?

Conclusions
Write your conclusions based on your results, the objectives of the lab practice and if your hypothesis
was accepted or not

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8. Mechanisms of Energy Generation: Cellular Respiration

Team:

Introduction

Plants are known for their ability to convert carbon dioxide into oxygen. However, all aerobic
organisms take in oxygen and give off carbon dioxide as long as they are alive. This is true for plants
as well as animals. During germination, seeds use sugars and other molecules as a substrate for
respiration.
Germination of a seed begins with water uptake by the seed. This process is called imbibition. The
uptake of water by a seed is an essential step in order for the seed to germinate. The total amount of
water taken up is about 2-3 times the weight of the seed. Whether or not a viable seed will germinate
depends on a number of factors. The chemical environment of the seed must be right. Water must be
available, oxygen has to be present since the seed must respire and no dangerous chemicals should
be present. The physical environment must also be favorable. The temperature must be suitable as
well as the light quality and quantity. The seeds are often buried underground. The reason is that this
helps guarantee the seeds receive the correct amount of light for germination. Full sunlight can often
prevent a seed from germinating. The extent to which germination has progressed can be determined
by measuring water uptake or respiration.
If you place a living organism in a closed system, it is possible to measure its consumption of oxygen,
or production of carbon dioxide. In this experiment, a respirometer can be used to measure oxygen
consumption, or Bromothymol Blue, an indicator of carbon dioxide may be used. As the seeds
consume oxygen, carbon dioxide is excreted. The carbon dioxide is then absorbed by the indicator,
creating a color change from blue to yellow. This color change will give an idea of how much oxygen
the germinating seeds consume.

Objectives
Students will:

● examine oxygen consumption by seeds

● understand the importance of respiration to life on Earth

PreLab Work
1. What do animals and plants need to live? What do they need to carry out cell respiration (to
breathe)?

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4. Fill in the table below with as many ideas as you can. The purpose of this activity is to find out
what you know before studying cell respiration, so don’t worry about being right or wrong.

What animals need?

What do animals and plants What do animals and plants give off?
need/use/consume? (reactants) (products)

5. Based on what you wrote above, write a summary equation for cellular respiration.

Hypothesis

Materials and Equipment

Hydrated Seeds
Sealable Bag
Bromthymol Blue, 0.04%
Sodium Hydroxide, 1%
Test Tubes
Rubber Stoppers
250-mL Flask
Test Tube Rack
Distilled Water
Paper Towel

Procedure

1. Two days before the experiment, rehydrate the seeds (wheat, barley, peas, or beans). Place
the seeds in a cup or beaker. Use dechlorinated water to cover the seeds to a depth at least 3
times their height in the container to compensate for the expansion of the seeds as they swell.
Allow the seeds to soak overnight.
2. Pour off the remaining water and fold the seeds into a wet paper towel. Place the towel in a
sealable bag. Close the bag and store the seeds in a dark place over a second night.
3. Prepare the bromothymol blue solution by adding 1.5 mL (about 30 drops) of 0.04%
bromothymol blue to 80 mL of distilled water. The prepared bromothymol blue solution should
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be green, but the shade of green may vary depending on the pH of your water source. A color
change will easily be observed if the solution given to students is slightly basic. To create a
slightly basic solution, add the sodium hydroxide dropwise to the bromothymol blue until the
color changes from green to a deep blue (10 to 20 drops).
4. The chemical bromothymol blue is an indicator that appears blue in an alkaline (base) solution
and yellow in an acidic solution. Carbon dioxide that is added to the solution surrounding the
seeds combines with water to form carbonic acid (H2CO3), turning the bromothymol blue to a
yellow-green color. Remove the carbon dioxide from solution and the bromothymol blue will
turn a deeper blue.
5. Fill a test tube ¾ full with rehydrated seeds. Pour the bromothymol blue solution over the seeds
until the tube is completely full. Seal the tube with a rubber stopper.
6. Prepare a control tube with only bromothymol blue solution. Fill the tube until it is completely
full, and seal it with a rubber stopper.
7. Store the samples until the end of the class period or overnight and have students make
observations regarding the color of the solution or any other changes they observe.

Observations and Results

Sketch your observations and describe them

Answer the following questions

● What do we call the process utilized by organisms to release energy which has been stored in
molecules like sugar?

● What is the equation for this energy releasing process?

● What causes the change in color of the bromothymol blue in the test tube?

● How does this relate to the rate of respiration?

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● Why can’t seeds just utilize photosynthesis to supply their energy needs?

● What is the selective advantage for seeds utilizing respiration?

● In which organelle does respiration take place?

Conclusions
Write your conclusions based on your results, the objectives of the lab practice and if your hypothesis
was accepted or not

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Bibliography

● Angulo, A.A., Galindo, A.R., Avendaño, R.C., Pérez, C. (2012). Biología Celular. Culiacán,
Sinaloa, México: Once Ríos.

● Alberts, B., Johnson, A., Lewis, J., Raff, M., Roberts, K., Walter, P. (2010). Biología molecular
de LA CÉLULA. Barcelona: OMEGA

● Atkins P, Jones L, (2006). Principios de Química: Los caminos del descubrimiento. Buenos
Aires: Médica Panamericana.

● Donald Voet, Judith G. Voet, Charlotte W. Pratt. (2007). Fundamentos de Bioquímica La vida a
nivel molecular. Buenos Aires: Médica Panamericana.

● Bewley, J.D., and M. Black. 1985. Seeds: Physiology of Development and Germination.
Plenum Press, New York.

● Cooper, E.L. 1997. Agriscience: Fundamentals & Applications. Delmar Publishers, Albany, New
York.

● Beller J. 1994. Low-Cost Biology Investigations. J Weston Walch, New York

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