Bioligirapport om kärsltrukturer och blod (EN)

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Biology 2
Laboratory report 4 - A bloody lab!



Authors:​ ​David Svensson and Moustafa Haydar Ahmad
Supervisor: ​Johan Lissel
Course​: ​Biologi 2
Date: ​From 2017-12-4 to 2018-01-22





Table of contents


1. Abstract​………………………………………………………………………………………………………..​1
2. Introduction​…………………………………………………………………………………………………...2
3. Material & Methods​…………………………………………………………………………………………..​4
3.1 Materials​4
3.2 Method​5
4. Results​………………………………………………………………………………………………………...​6
4.1 Wright’s stain experiment​…………………….…………………………………………………..​6
4.2 Blood Sugar experiment​………………………………………………………………………….​8
4.3 Heart Dissection​………………..…….……………………………………………………………8
4.4 Blood pressure and heart rate experiment​………….…………………………………………​1​0
4.5 Time for blood circulation​………………………………………………………………………..​1​1
5. Discussion​…………………………………………………………………………………………………...​1​1




1.

Abstract


The aim of the laboratory experiments was to examine:

●White blood cells in proportion to red blood cells in a laborant and to calculate how
long it took the blood to circulate in the laborants​ body
●How the blood sugar levels of a laborant varied during a timed period
●The internal structures of a mammal heart
●A laborant’s blood pressure, heart rate and electrocardiogram (ECG)

For the first experiment, a Wright stain protocol was followed in order to examine white blood
cells, which resulted in the discovery of several white blood cells such as lymphocytes,
neutrophils, monocytes and eosinophils. The second experiment ​began with a​ volunteer
consuming a carbohydrate-rich meal and then their blood sugar levels were measured in
regular intervals. The blood sugar level of the volunteer peaked at 8,2 mmol/L. ​During the
third experiment, a mammal heart was dissected and its various structures were examined​.
A few of the structures identified were the epi-myo, endo-cardium, the aorta and the
trabeculae carneae. ​Under the duration of the fourth experiment, a laborant’s blood pressure
and heart rate were measured and an electrocardiogram was analysed. ​Laboratory
equipment such as a Sphygmomanometer (blood pressure monitor) and the “Coala Life”
heart rate monitor were used during these experiments.

1




2.

Introduction

Blood cells & blood circulation

Blood is an essential body fluid for most organisms in the animal kingdom, and without it
many species of animals could not be able to live. Blood is mostly comprised of blood cells
and blood plasma, however, in this introductory part, the main focus is on the white and red
blood cells as well as the time for blood circulation in the human body.[1]

Red blood cells are the most typical kind of blood cell in the human body, and ​their​ primary
function is, through the circulatory system, to deliver oxygen to cells. These red blood cells
absorb oxygen in the lungs and then releases it to the cells of the body through the
circulatory system. The red blood cells have an abundance of hemoglobin, which ​enables it
to​ bind ​with​ oxygen and to possess a blood red colour.[2]

White blood cells on the other hand, are a less typical kind compared to the red blood cells.
These type of cells are cells of the immune system and responsible for protecting the body
from harmful diseases and infections. ​White blood ​cells are produced by the hematopoietic
cells in the bone marrow. A big difference which separates white blood cells from other blood
cells is the presence of a nuclei, which the white blood cell possesses. These nuclei come in
different shapes and forms, which dictate their appearance and function. Depending on the
nuclei and granules, a white blood cell may target parasites in the body or release histamine
for inflammatory responses. The different types of white blood cells are: neutrophils,
eosinophils, basophils, lymphocytes and monocytes. [3][4]

The difference in abundance of white compared to red blood cells is very large, the ratio of
white blood cells compared to red blood cells being 1:400 up to 1:1000. [4]

The human male body contains circa 5 litres of blood, and in order to calculate the correct
amount of time for blood in the human male body to circulate, the heart’s stroke volume as
well as the heart’s BPM have to be known. The stroke volume can vary from 50 ml per
stroke up to 90 ml per stroke. [5] A simple equation for the amount of time for the blood to
circulate can be formulated as: Time (Minutes) = . This equation is supported
Volume of blood
BPM × Stroke volume

by the logical reasoning of if the amount of beats per minute increases, the time for a total
circulation decreases. The same principle applies for the stroke volume.


Heart

Organisms have evolved multiple ways of transporting nutrients and metabolic waste in the
body, the most common transportation organ that have evolved in animals is the heart. The
heart is a very important muscular organ in the animals that have them, and its function is to
pump blood in the pulmonary circulatory system and cardiovascular circulatory system. The
blood is first pumped to the pulmonary circulatory system where it becomes enriched with
oxygen (through the process of respiration) from the lungs and is then transported to the
cardiovascular system, which assists the body with the transportation of oxygen but also
nutrients, the immune system, erythrocytes (red blood cells), hormones and the removal of
metabolic waste. The heart, in humans, is located between the lungs. In mammals (including
2



humans) and birds, the heart is designed to have four chambers: the upper left and right
atria; and the lower left and right ventricles.[6]

The heart rate depends of the activity of the organism as it increases with intense physical
activity and decreases while resting, but other factors such as fitness, age, gender and
physical size also affects the heart rate. In most healthy human adult males (18-25 years)
the average resting heart rate is 70-73 BPM (beats per minute) while in adult human females
(18-25 years) the average resting heart rate is 74-78 BPM. [7]


Blood pressure

Blood pressure is the pressure of the blood vessel walls which arises when blood circulates
through them. Systolic and diastolic blood pressure are terms for how the maximum
pressure is during one heartbeat and minimum pressure between two heart beats
respectively. The pressure is measured in mmHg, and is often abbreviated 120/80 if the
systolic pressure is 120 mmHg and the diastolic pressure is 80 mmHg. [8] Many factors can
affect blood pressure, such as physical fitness and activity, heart diseases, emotions and
diet. [9][10]


Blood sugar

Blood sugar levels describe the amount of glucose in the blood of humans and other
animals. The body strictly regulates the simple sugar glucose (for the body to have circa 4
grams at all times) as a part of metabolic homeostasis. The hormone which primarily
regulates the uptake of glucose is insulin, which is produced in the pancreas. Blood sugar
levels are most often lowest in the morning, and then rises after consumed meals, especially
if they are high in glucose. [11][12]


ECG

An ECG (electrocardiography) is process of recording the electrical signals in a heart over a
certain period of time. Each beat of the heart is triggered by special pacemaker cells in the
upper right chamber of the heart. An electrocardiogram is a diagram which records the
timing and voltage of the electrical signals as they travel through the heart. [13][14]









3





3.

Material & Methods

3.1 Materials



Laboratory equipment

Chemicals & Miscellaneous


Wrights stain experiment


1.Microscope slide
2.Disinfectant napkin
3.Bunsen burner
4.Matches
5.Forceps
6.Microscope
7.Blood lancet
8.Paper towels



Blood sugar experiment




1.Glucose meter
2.Disposable test strip
3.Blood lancet
4.Timer





Heart dissection


1.Heart (pig)
2.Scalpel
3.Pair of medical latex gloves
4.Dissection probe
5.Paper towels
6.Plastic tray
7.Tweezers


Blood pressure and heart rate experiment


1.Sphygmomanometer (blood pressure
monitor)
2.Coala Life heart rate monitor (ECG)



Wrights stain experiment


1.Wright’s stain
2.Deionized water
3.Pair of medical latex gloves
4.Blood
5.Tape






Blood sugar experiment


1.Gingerbread cookies
2.Julmust
3.Plastic mugs
4.Paper towels





Heart dissection


1.Assistant
2.Picture of mammal heart structures
3.Camera






Blood pressure and heart rate experiment


1.Notebook
2.Timer
4







3.2 Method



Wright’s stain experiment


1.Two microscope slides were cleaned and prepared and the ends of each slide were
taped
2.Volunteer finger was disinfected with a disinfectant napkin
3.A blood lancet was used to pierce the skin of the finger
4.Volunteer slowly placed their finger on the microscope glass
5.A microscope slide was placed on top of the blood drop, and a thin layer of the blood
was smeared across the microscope slide
6.The microscope slides were air-dried for circa 10 minutes
7.Four drops of Wright’s solution were added to the microscope slides
8.A bunsen burner was turned on and lit with matches
9.The slides were put above the flame for 2-3 seconds to fix the blood smear on the
microscope slides
10.The microscope slides were carefully washed with deionized water
11.The microscope slides were air-dried for another 10 minutes
12.The blood on the microscope slides was examined to document any white blood cells
with the use of a microscope with ​1000x​ magnification


Blood Sugar experiment

(​Prior to experiment-​ The test subject must not have consumed any food, so blood sugar levels can
be studied easily.)

1.The test subjects finger was pierced with a blood lancet and blood was transferred on
a disposable test strip
2.Initial blood sugar levels of the test subject was measured and documented
3.Subject gorges in carbohydrate-rich consumables such as gingerbread cookies and
julmust between test intervals
4.A timer was used to measure the blood sugar levels in regular intervals (15 min each
interval, up to an hour)


Heart Dissection


1.Dissection equipment such as dissection probes, paper towels, a plastic tray, a
scalpel, and tweezers were prepared
2.The pig heart was placed on top of the tray
3.Coronary veins were photographed
4.The heart was flipped and the top of the heart was photographed
5.The heart was cut along the coronary vein to expose its internal structures
6.Dissecting probes were used to pin heart in place
5



7.Internal structures were photographed
8.The heart was disposed into the trash and used equipment was cleaned after
finishing the experiment


Blood pressure and heart rate experiment


1.A Sphygmomanometer (blood pressure monitor) was placed on the left brachial
artery and was started
2.Initial blood pressure and heart rate were measured and documented
3.A timer was used to measure the blood pressure and heart rate in regular intervals
(15 min each interval, up to 45 min)




4.

Results



4.1 Wright’s stain experiment






Picture 1: A picture from a microscope that shows 3 different types of white blood cells ​(1000x magnification)


6







Picture 2: A picture from a microscope that shows a monocyte white blood cell ​(1000x magnification)








Picture 3: A picture from a microscope that shows a eosinophil white blood cell ​(1000x magnification)


7





4.2 Blood Sugar experiment



Graph 1: A graph (with a function) showing the changing values of blood sugar levels during a period of 60 minutes​ (E
in the function is ten to the power of)



4.3 Heart Dissection



Picture 4: Exterior of heart with some veins and structures highlighted in color

8




Picture 5: Top of the heart showing major arteries and veins highlighted in color

Picture 6: A dissected heart showing the inside structures and parts of the heart highlighted in color

9




Picture 7: The left atrium of the dissected heart and other parts are highlighted in color



4.4 Blood pressure and heart rate experiment



Graph 2: Graph (with 3 functions) showing how pulse, systolic and diastolic pressure varied under a period of 45
minutes ​(E in the function is ten to the power of)

10







Picture 8: An electrocardiogram showing the electrical activity of a laborant’s heart



4.5 Time for blood circulation

Using the equation , presuming that the volume of blood of an adult male
Volume of blood
BPM × Stroke volume

human to be 5 L, taking the average BPM from graph 2, and also presuming that the stroke
volume is 70 ml/beat, the result is ~1 minute. This means that the blood of an adult male
human (with the chosen BPM and stroke volume) requires circa 1 minute in order to circulate
the whole body.


5.

Discussion



Wright’s stain experiment

The first goal of this experiment was to stain white blood cells purple while leaving red blood
cells to be transparent, and this was done in order to see (with a microscope) different types
of white blood cells. The second goal of this experiment was to determine the ratio of white
blood cells compared to red blood cells, and this is achieved by taking a picture through the
eyepiece lense. This experiment was performed several times until the expected results
were achieved, because the most common problem which hindered optimal results was the
blood being too dense for proper analysis of the nucleus of white blood cells and the ratio of
the white and red blood cells.

11



In picture 1, two lymphocytes and a neutrophil ​were identified​ due to the shape of their
nuclei. In picture 2, it can be seen that the single white blood cell, although difficult to
distinguish from other similar types white blood cells, to be a monocyte. In picture 3, the
white blood cell was easily determined to be an eosinophil due to its characteristic nucleus.

The ratio of white to red blood cells, according to sources, should be between 1:400 to
1:1000. After counting the red (transparent) blood in picture 2, it was determined that the
amount of red blood cells was 390, which means that the ratio of white to red blood cells is
1:390. Even though this means that the participant that donated their blood for this
experiment had a higher ratio of white blood cells compared to red blood cells that the
source specified, an error source could be that more samples are required to be studied in
order to determine a more accurate ratio. The analysing of more samples is however
hampered due to the copious amount of red blood cells in every sample taken, making it
difficult to count them without the use of a machine/program. The probability of the ratio
1:390 appearing could be high due to the participant being slightly sick, meaning that the
body produces more white blood cells than normal, which could explain why the ratio of
white blood cells is higher than the ratios given by the source. [15]


Blood sugar experiment

The blood sugar experiment was performed in order to observe how blood sugar levels
varies when a test subject does not eat breakfast and then consumes a carbohydrate-rich
meal. During 60 minutes, the subject’s blood sugar level is measured every 15 minutes in
order to observe how a carbohydrate-rich meal changes the blood sugar levels. In graph 1, it
can be seen that the blood sugar level is very low in the beginning, and then increasing
because of the ingested carbohydrates. The blood sugar level reaches its peak at 8,2
mmol/L 30 minutes in, and then declining afterwards and stabilising around 7 mmol/L. A
function with the R2 value of 1 (an accurate function) was created by a program in order to
see a curve which describes the change of the blood sugar levels in this particular period of
time. The function should however not used outside the timeframe. For example, if the
variable of the function is to exceed 60 minutes, the calculated blood sugar level would
increase and result in overly absurd values.


Heart dissection

The aim of this experiment was to dissect a mammal heart while examining and
documenting the structures. A pig's heart was cut open using dissection tools such as
scalpels. Various structures of the heart were photographed. Laborants used assistive tools
such as a video dissecting guides and a picture of the mammal heart ​[picture]​ for identifying
the different structures of the heart. The experiment was performed once and the heart was
disposed of after experimentation. ​The online photo editing tool Pixlr was used to highlight
the hearts various structures.

Picture 4 shows the exterior view of the heart that contains some identifiable structures. The
picture shows the shape of the apex, the bottom of the heart and and the heart's coronary
blood vessels which are the blood vessels of the heart muscle (myocardium) alongside
minor fat deposits. On the top of the heart the atrium is slightly visible. We decided to cut the
12



heart vertically along the coronary blood vessels because we hypothesised that it was the
most effective way to cut it open without damaging any of the internal structures.

Picture 5 shows the top of the heart, mainly where the great blood vessels are located. The
Aorta (in pink) was identified as well as the pulmonary trunk/artery (in yellow), pulmonary
vein (in blue) and the vena cava (in green). This task was made difficult mainly because the
major blood vessels had been cut short and therefore much harder to identify. The idea of
some of the blood vessels not being identified correctly has not been dismissed and the
errors may be reflected in the results.

Picture 6 shows the results of the vertical dissection and some of the internal structures were
able to be identified by using a picture-guide on the heart. The heart was fresh, and
therefore soft and asymmetrically shaped, which hampered our ability to identify structures
as effectively as we could if we had a more rigid heart. Similarly to the blood vessels, the
idea of some of the blood vessels not being identified correctly has not been dismissed and
the errors may be reflected in the results. Some identified structures were the Epi-Myo and
Endo-cardium (in green) which together are the heart's walls. The heart's upper chambers,
the left and the right atrium were identified (in yellow and red respectively). While the heart is
beating, the atria receives blood, and when the heart muscle contracts, it pumps the blood
into the ventricles. The ventricles are marked as RV and LV. The left ventricle is responsible
to pump blood to the cardiovascular circulatory system so nutrient- and oxygen-filled blood
reaches the body while the RV pumps blood to the pulmonary circulatory system so the
blood can become oxygenated by respiration.

Other structures identified in picture 6 includes the trabeculae carneae (in orange) a internal
structure that consists of muscular ridges that covers all of the inner wall. According to
sources the inner walls evolved to be rigid because had the inner walls been flat there would
be suction and thus impair the heart's ability to pump efficiently. The chordae tendineae
structure (in blue) was also identified and can be described as a “string-like” structure. The
function of the trabeculae carneae is to control the blood flow from the atrioventricular valves
by tensing or relaxing, to regulate blood flow. [16]

In picture 7 the papillary muscles were identified and they can be described as very thick
muscular ridges. This muscles are connected to the trabeculae carneae. According to
sources, the papillary muscles of the left ventricle attach via chordae tendineae to the mitral
valve, which controls the flow from the left atrium to the left valve. [17]



Blood pressure & heart rate experiment

The goal of the blood pressure & heart rate experiment was to observa how blood pressure
varies under a period of 45 minutes and also observe how the heart rate varies during this
period. In graph 2, the function which describe the systolic and diastolic pressure as well as
the heart rate has an R2 value of 1, which means that the function is accurate. The rule of
not using the function outside the recorded period of time also applied here, seeing how the
values would continue to increase or decrease without stabilising. The functions of the
systolic and diastolic pressure both form curves which are identical to each other, meaning
13



that there could be a possible correlation between systolic and diastolic pressure, however
there is not enough data to support this theory.


ECG

In picture 8, a electrocardiogram shows the voltage over time of a volunteer during a 30
second period. During this period, only 4 irregularities occurred which all can be attributed
(due to it only happening in the beginning of the diagram) to the volunteer adjusting their
finger on the ECG device.


6. Source citation and criticism



Digital Sources

Wikipedia


[3]​


https://en.wikipedia.org/wiki/White_blood_cell

[6] ​https://en.wikipedia.org/wiki/Heart
[9] ​https://en.wikipedia.org/wiki/Blood_pressure
[11] ​https://en.wikipedia.org/wiki/Blood_sugar_level
[14] ​https://en.wikipedia.org/wiki/Electrocardiography
[16] ​https://en.wikipedia.org/wiki/Trabeculae_carneae

Various sources from Wikipedia were used during the beginning to research the yeast experiment.
Wikipedia was mainly used to find other more trustworthy ​
primary sources
​, such as academic
university sources. Most of the academic sources that have been gathered from Wikipedia cannot be
easily altered which adds to their credibility. The little information that has been directly taken from
Wikipedia has been compared to other sources, and has been deemed credible because the
information they provide are similar to each other.



Universities/Academic Sources


[2] ​http://www.hematology.org/Patients/Basics/
[5] ​http://www.lissel.info/n15a,en,blodig,lab,5,dec,2017.html


[12] ​https://www.hsph.harvard.edu/nutritionsource/carbohydrates/carbohydrates-and-blood-sugar/
[17] ​http://www.vhlab.umn.edu/atlas/right-ventricle/papillary-muscles/index.shtm

The credibility of the academic sources is considerably higher than non academic sources (such as
news) because they have been ​
peer-reviewed
​and published on their respective universities website.
Universities try to keep their academic reputation and trustworthiness high by avoiding spreading
misinformation that could tarnish it. Thes...

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  • Moustafa Haydar Ahmad 2018-05-15

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