REMOTE LAB ACTIVITY
SUBJECT SEMESTER: ________________
TITLE OF LAB: Diseased Cell
Lab format: This lab is a remote lab activity.
Relationship to theory (if appropriate): In this lab you will identify the underlying cellular changes that
lead to several human diseases.
Instructions for Instructors: This protocol is written under an open source CC BY license. You may use
the procedure as is or modify as necessary for your class. Be sure to let your students know if they
should complete optional exercises in this lab procedure as lab technicians will not know if you want
your students to complete optional exercise.
Instructions for Students: Read the complete laboratory procedure before coming to lab. Under the
experimental sections, complete all pre-lab materials before logging on to the remote lab, complete
data collection sections during your on-line period, and answer questions in analysis sections after your
on-line period. Your instructor will let you know if you are required to complete any optional exercises
in this lab.
Remote Resources: Primary - Microscope; Secondary - Diseased Cell Slide Set.
CONTENTS FOR THIS NANSLO LAB ACTIVITY:
Learning Objectives..........................................................................................................
Background Information .................................................................................................
Equipment .......................................................................................................................
Experimental Procedure .................................................................................................
Pre-lab Exercise 1: Skin Composite and Skin Cancer Slides Observations .....................
Exercise 1: Skin Composite and Skin Cancer Slides Observations .................................
Pre-lab Exercise 2: Normal Red Blood Cells and Sickled Cells Slide Observations .........
Exercise 2: Normal Red Blood Cells and Sickled Cells Slide Observations .....................
Pre-lab Exercise 3: Normal and Cancerous Breast Tissue ..............................................
Exercise 3: Normal and Cancerous Breast Tissue ..........................................................
Pre-lab Exercise 4: Normal and Cancerous Spleen ........................................................
Exercise 4: Normal and Cancerous Spleen .....................................................................
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9 - 10
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10 - 11
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11 - 12
12
12 - 13
IF CARDIAC IS USED IN ADDITION TO OR IN PLACE OF SPLEEN ADD
Pre-lab Exercise 4A: Cardiac Muscle and Myocardial Infarction .................................... 13
Exercise 4A: Cardiac Muscle and Myocardial Infarction ................................................ 13
Pre-lab 5: Cirrhosis of the Liver ...................................................................................... 14
Exercise 5: Cirrhosis of the Liver .................................................................................... 14
Summary Questions: ....................................................................................................... 14 - 15
Preparing to Use the Remote Web-based Science Lab (RWSL) ...................................... 16 - 30
LEARNING OBJECTIVES:
After completing this lab, you should be able to do the following things:
1.
2.
3.
4.
Identify the differences between normal skin tissue layers and skin cancer.
Compare the shape of normal red blood cells and sickled red blood cells.
Identify the cellular differences between normal breast tissue and breast cancer.
Identify the cellular differences between a normal spleen and a spleen suffering the effects of
leukemia.
AND/OR – Use the following objective if you want to use cardiac muscle exercise in
addition to spleen or to replace spleen
Identify and describe the difference between healthy cardiac muscle and cardiac muscle
suffering the effects of myocardial infarction.
5. Identify the cellular changes that occur in the liver due to cirrhosis.
BACKGROUND INFORMATION:
Cells inside organisms undergo a variety of changes as part of adaptations due to growth, stress, and
aging. During these adaptations, cells are trying to maintain homeostasis to prevent diseases. The
human body can be affected by several stimuli such as:
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Physical agents (mechanical trauma, temperature variation).
Chemical agents (radiotherapy, glucose, or lipid accumulation).
Infectious microorganisms (toxins produced by bacteria or viruses interfering with normal cell
metabolism).
Hypoxia: lack of oxygen supply to cells (it could be detrimental for the brain).
Genetic factors leading to abnormal cellular metabolism or malformation.
Nutritional imbalances (more frequent in children).
Hypersensitivities and allergic reactions
However, some of the changes could cause irreversible effects leading to pathological changes that
could either be obvious or difficult to detect. Many of the common diseases that affect humans are
caused by disruptions of the basic mechanisms of the cell. Medical professionals and scientists rely on
histopathology, the microscopic study of tissue, to accurately diagnose diseases of the human body.
During this laboratory activity, you will be viewing and comparing different types of cells and tissues to
identify the differences between diseased and normal tissues at the cellular level. We will be looking at
cells from several different tissues in the body including the skin, internal organs, and the blood. Each
tissue type is described in detail below.
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Skin
Our skin is the largest organ. It has several functions: serving as a protective barrier; regulating our body
temperature; producing sweat; and other functions. Although most diseases affecting the skin originate
in the layers of the skin (Figure 1), such abnormalities are also important factors in the diagnosis of a
variety of internal diseases. There is some truth in the belief that the skin mirrors a person’s internal
health. Often, the visibility and accessibility of skin make it the first organ of the body to show
detectable signs of underlying disease.
Figure 1: Normal human skin tissue specimen
Normal human skin is composed of three layers: epidermis; dermis; and hypodermis. The epidermis is
the outermost layer. The epidermis and dermis together make up the cutis. The hypodermis makes up
the subcutis and contains structures like the hair follicles and sweat glands.
Abnormalities of the skin frequently suggest metabolic, malignant, and glandular diseases. Skin cancer is
the most common form of cancer in the United States. The two most common types of skin cancer—
basal cell and squamous cell carcinomas—are highly curable. However, melanoma, the third most
common skin cancer, is more dangerous. About 65%–90% of melanomas are caused by exposure to
ultraviolet (UV) light (http://phil.cdc.gov/phil/details.asp.)
UV light is not the only cause of skin cancer. Kaposi sarcoma (Figure 2) is a malignant tumor of the
lymphatic endothelium caused by the Human herpesvirus 8 (HHV8), i.e., Kaposi’s sarcoma-associated
herpesvirus (KSHV) and arises from a cancer of the lymphatic endothelial lining. It is characterized by
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bluish-red cutaneous nodules. Kaposi’s sarcoma is thought of as an opportunistic infection affecting
patients whose immune systems have been compromised, as in the case of patients with HIV/AIDS
(http://phil.cdc.gov/phil/details.asp.)
Figure 2: Human skin biopsy specimen due to Kaposi's sarcoma
Above is a photomicrograph depicting the histopathologic changes seen in human skin biopsy specimen
due to Kaposi’s sarcoma. Of importance are the appearance of the dermal layer which contains a cellular
infiltrate and a proliferation of vascular elements.
Like other tissues, skin is afflicted by all types of pathological changes, including hereditary,
inflammatory, benign and malignant (neoplastic), endocrine, hormonal, traumatic, and degenerative
processes. Emotions affect the health of the skin as well. The reaction of the skin to certain diseases and
disorders differs from that of other tissues in many ways. For example, extensive inflammation of the
skin may affect metabolism within other organs and systems of the body, causing anemia, circulatory
collapse, disorders of body temperature, and disturbance of water and electrolyte balance in the blood
(http://phil.cdc.gov/phil/details.asp.)
Internal Organs
The internal organs of a body are complex structures consisting of a combination of parenchymal tissue
and stromal tissue. The cells of the parenchyma are specialized to carry out the function of the organ,
while the stromal cells are involved in support (e.g. connective tissue and blood vessels).
For example, the microscopic anatomy of the liver reveals a uniform structure of clusters of cells
called lobules where the vital functions of the liver are carried out. Each lobule, measuring about
one millimeter in diameter, consists of numerous cords of rectangular liver cells, or hepatocytes,
that radiate from central veins, or terminal hepatic venules, toward a thin layer of connective
tissue that separates the lobule from other neighboring lobules.
Hepatocytes occupy about 80 percent of the volume of the liver, and their cytoplasm (the area
surrounding the nucleus) contains many mitochondria which provide the energy needed for the many
synthetic and metabolic functions of the liver cell. The cytoplasm also contains a series of long tubules
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called the endoplasmic reticulum which provides many enzymes essential to liver function. Some of the
membranes of the endoplasmic reticulum appear granular, or rough, owing to the presence of
ribosomes which are responsible for forming specific polypeptide (protein) chains after having had the
amino group removed (deamination) and having been converted into glucose through a process called
gluconeogenesis. The ammonia released from gluconeogenesis is converted to urea in the hepatocyte
by way of the urea cycle. The nonribosomal, or smooth, endoplasmic reticulum is where cytochromes
(combinations of heme from hemoglobin with various proteins), and certain enzymes undertake the
important hepatic functions of drug and hormonal metabolism and also cholesterol synthesis.
Hepatocytes also conjugate with carbohydrate components of bilirubin and other fat-soluble metabolic
and foreign compounds and thereby are made soluble in water. Bilirubin is the product of hemoglobin
metabolism that is formed in the bone marrow and the lymphatic tissue and is carried to the liver after
becoming bound to plasma albumin. It is released at the hepatocytic sinusoidal membrane and is
transported to the smooth endoplasmic reticulum where it is conjugated with one or two molecules of
glucuronic acid and thereby becomes soluble in water and excretable in bile. The Golgi apparatus, a
series of tubular structures between the endoplasmic reticulum and the canaliculus, acts as a transport
station for newly made proteins and other hepatocytic products before they are conveyed to other parts
of the cell or out of the cell entirely. Lysosomes, another important cytoplasmic constituent, are
responsible for the intracellular storage of pigments, such as iron or copper, and for the digestion of
certain contents, such as glycogen or foreign particles. The nucleus of the hepatocyte guides replication
of the cell and transmits genetic material in the form of messenger ribonucleic acid (mRNA) from
deoxyribonucleic acid (DNA) to organelles located in the cytoplasm (http://phil.cdc.gov/phil/details.asp.)
The parenchymal cells of certain organs, such as the liver, the heart, and the kidneys, are responsible for
the metabolism and elimination of excess fat. If the fat droplets are not properly metabolized, they end
up accumulating in the endoplasm reticulum and the Golgi apparatus of the cells (Figure 3). This
accumulation could be irreversible, leading to more serious health problems, and may even lead to
death.
Figure 3: Alcoholic Cirrhosis Liver
Above is an image of a liver with alcoholic cirrhosis. The yellow color indicates the accumulation
of fat inside the hepatocytes interfering with normal activity of these cells due to excessive
alcohol consumption.
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References
Encyclopedia Britannica
http://www.britannica.com/EBchecked/topic/1081754/human-digestivesystem/242929/Microscopic-anatomy#ref212928
Blood and the Circulatory System
The body’s circulatory system is composed of several parts: the blood vessels; the blood; heart; and the
lungs. The main function of the circulatory system is to transport food and oxygen to the body’s cells
and remove waste products and CO2. The structure of the blood vessels, both large and small, is closely
associated with their function (Figure 4). All nutrient materials and waste products exchanged between
the organs and the blood must traverse perivascular spaces in the walls of the vessels that are occupied
by connective tissue. One of the important functions of the connective-tissue cells is to maintain
conditions in the extracellular spaces that favor this exchange
(http://www.britannica.com/EBchecked/topic/36874/artery#md-media-strip-tab-image-content.)
Figure 4: Artery: transverse section of an artery
http://www.britannica.com/EBchecked/topic/36874/artery#md-media-strip-tab-image-content
Blood vessels are composed of multiple layers of tissue. Each of these layers provides specific functions
to the vessel. Connective tissue makes up the outer surface of the vessels and controls transport into
and out of the vessel. The elastic membrane and elastic fibre give the vessel its flexibility while the
involuntary muscles help to maintain blood pressure and the endothelium forms the inner surface of the
vessel.
Any variation in the diameter of the lumen of the blood vessels will have an impact on the blood flow,
nutrients supply, blood pressure, and the overall function of the cardiovascular system.
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The blood vessels provide the pathway through which the blood circulates. The blood is composed four
different types of cells: red blood cells (erythrocytes); white blood cells (leukocytes); and platelets
(fragment of giant cells also known as thrombocytes). Each one of these cells is responsible for specific
functions inside our organs. Any variation of the number, shape, or size of these cells will have an impact
on our health. The function of these cells is:



Red blood (erythrocytes) cells function in the transport of oxygen and carbon dioxide to and
from the cells.
White blood (leukocytes) cells are involved in the immunity.
Platelets are involved in hemostasis (stopping the bleeding).
ADD IF CARDIAC OBJECTIVE IS USED
Cardiac Muscle
The heart is the pump that keeps blood circulating throughout the body and thereby transports
nutrients, breakdown products, antibodies, hormones, and gases to and from the tissues. The heart
consists mostly of muscle. The myocardial cells (collectively termed the myocardium) are arranged in
ways that set it apart from other types of muscle. The outstanding characteristics of the action of the
heart are its contractility, which is the basis for its pumping action, and the rhythmicity of the
contraction.
Heart muscle differs from its counterpart, skeletal muscle, in that it exhibits rhythmic contractions. The
amount of blood pumped by the heart per minute (the cardiac output) varies to meet the metabolic
needs of the peripheral tissues (muscle, kidney, brain, skin, liver, heart, and gastrointestinal tract.) The
cardiac output is determined by the contractile force developed by the muscle cells of the heart
(myocytes) as well as by the frequency at which they are activated (rhythmicity.) The factors affecting
the frequency and force of heart muscle contraction are critical in determining the normal pumping
performance of the heart, and its response to changes in demand.
The heart is a network of highly branched cardiac cells 110 μm in length and 15 μm in width which are
connected end to end by intercalated disks. The cells are organized into layers of myocardial tissue that
are wrapped around the chambers of the heart. The contraction of the individual heart cells produces
force and shortening in these bands of muscle with a resultant decrease in the heart chamber size and
the consequent ejection of the blood into the pulmonary and systemic vessels.
Your heart muscle needs oxygen to survive. A heart attack occurs when the blood flow that brings
oxygen to the heart muscle is severely reduced or cut off completely. View an animation of blood flow.
This happens because coronary arteries that supply the heart muscle with blood flow can slowly become
narrow from a buildup of fat, cholesterol, and other substances that together are called plaque. This
slow process is known as atherosclerosis. In a heart artery, when plaque breaks, a blood clot forms
around the plaque. This blood clot can block the blood flow through the heart muscle. When the heart
muscle is starved for oxygen and nutrients, it is called ischemia. When damage or death of part of the
heart muscle occurs as a result of ischemia, it is called a heart attack or myocardial infarction (MI).
About every 34 seconds, someone in the United States has a myocardial infarction (heart attack).
END OF TEXT IF CARDIAC OBJECTIVE IS USED
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Reference:
Encyclopedia Britannica
http://www.britannica.com/EBchecked/topic/132995/connective-tissue
ADDITIONAL REFERENCES IF CARDIAC IS USED
About Heart Attacks, American Heart Association, last reviewed on 10/20/2013.
http://www.heart.org/HEARTORG/Conditions/HeartAttack/AboutHeartAttacks/About-HeartAttacks_UCM_002038_Article.jsp
EQUIPMENT:
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Paper
Pencil/pen
Slides
o Skin Cancer, Section
o Adult Skin, Unpigmented, section
o Human Sickle Cell Anemia, Smear
o Human Blood, Smear
o Leukemia Spleen, Section
o Spleen, Section
o Myocardial Infarction, Section (IF CARDIAC IS USED)
o Cardiac Muscle, Section (IF CARDIAC IS USED)
o Breast Carcinoma, Section
o Mammary Gland, Inactive, Section
o Cirrhosis, Section
o Liver, Section
Computer (access to remote laboratory and for data analysis)
EXPERIMENTAL PROCEDURE:
Once you have logged on to the remote lab system, you will perform the following laboratory
procedures. See Preparing for the Microscope NANSLO Lab Activity below.
PRE-LAB EXERCISE 1: Skin Composite and Skin Cancer Slides Observations
In this exercise, you will use the microscope to distinguish between the three layers of human skin. In
this exercise, you will identify the skin layers on images you have taken, and you will measure and
compare the relative size and shape of cancerous skin cells with healthy skin cells.
Pre-Lab Questions:
1. Using what you know about cancer, do you think the size and shape of the cancerous skin cells
will be bigger, smaller, or the same as the ones of the healthy skin?
2. Rewrite your answer to question one in the form of an If … Then … hypotheses.
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EXERCISE 1: Skin Composite and Skin Cancer Slides Observations
Data Collection:
3. Select the composite skin slide (Slide Cassette 4: TBA) from the slide loader. Using the 10X
objective, identify the skin sample and bring it in to focus.
4. Carefully work your way through all the objectives focusing with each one until you reach the
60X objective and capture an image. Insert your image of normal skin in the space below.
5. Select the skin cancer slide (Slide Cassette 4: TBA) from the slide loader. Using the 10X
objective, identify the skin sample and bring it in to focus.
6. Carefully work your way through all the objectives focusing with each one until you reach the
60X objective and capture an image. Insert your image of skin cancer in the space below.
Analysis:
7. Using the image you took in question 4 of this exercise, label the different layers of the skin and
the 3 different parts of a single cell (nucleus, cytoplasm and cell membrane) from either the
epidermis or the dermis. Insert your labeled image below.
8. Using the image you took in question 6 of this exercise, label the different layers of the skin and
the 3 different parts of a single cell (nucleus, cytoplasm and cell membrane) from either the
epidermis or the dermis. Insert your labeled image below. Describe the shape of the cell and its
nucleus.
9. Based on your observation, describe the difference in the shape and number of cell layers
between normal skin and skin cancer.
10. Next we are going to measure the size of normal and cancerous cells. To determine the size of
the cells, we are going to use the ratio method. In order to do this, you will need one piece of
information which is the width of your field of view. On our microscopes, the field of view at
60X is 205µm.
11. Now, if we use the image in figure 5, we can see that the total width of the field of view is 13.6
cm or 136 mm (Image A). The cell (Gray) is 3.7 cm or 37mm (Image B).
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Figure 5: Measurements
12. Now, if we divide 37mm/136mm = 0.272, which we multiply by the total length of the field of
view. So 0.272 * 205µm = 55.77 µm rounded for significant figures, we get a cell size of 56µm.
13. Measure the difference in the size of the healthy skin and compare that to the size of the
cancerous skin.
14. Are your results in correlation with what you have predicted earlier?
15. Rewrite your hypothesis in light of the new information you collected in this exercise.
PRE-LAB EXERCISE 2: Normal Red Blood Cells and Sicklled Cells
Slide Observations
The function of red blood cells is to transport oxygen to cells and carbon dioxide away from them. The
shape and size of the red blood cell is important for its function. In this lab exercise, you will measure
and compare the relative size and shape of normal red blood cells with the sickled celled ones.
Pre-lab Questions:
1. Do you predict the size of normal red blood cells to be smaller, bigger, or the same as the sickled
celled one?
2. Rewrite your answer to question one in the form of an If … Then … hypotheses.
EXERCISE 2: Normal Red Blood Cells and Sickeled Cells Slide Observations
Data Collection:
3. Select the human blood slide (Slide Cassette 4: TBA) from the slide loader. Using the 10X
objective, identify the blood cells and bring them into focus.
4. Carefully work your way through all the objectives focusing with each one until you reach the
60X objective and capture an image. Insert your image of normal red blood cells below.
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5. Select the sickle cell anemia slide (Slide Cassette 4: TBA) from the slide loader. Using the 10X
objective, identify the blood cells and bring them in to focus.
6. Carefully work your way through all the objectives focusing with each one until you reach the
60X objective and capture an image. Insert your image of sickled cells below.
Analysis:
7. Using the method from exercise 1, determine the length of the normal and sickle red blood
cells.
8. Based on your observation, describe the difference in the shape of normal red blood cells versus
sickled cells.
9. Are your results in correlation with what you have predicted earlier?
10. Rewrite your hypothesis to take into account the new information you have learned in this
exercise.
11. What is the impact of sickle cell anemia on oxygen transport?
PRE-LAB EXERCISE 3: Normal and Cancerous Breast Tissue
As we mentioned in experiment 1, there are several forms of cancer. The most common form of cancer
in women is breast cancer. Rather than being a cancer of the epithelial tissue, breast cancer is a cancer
of the mesenchymal tissue. In this exercise, you will compare and identify the differences between
normal and cancerous breast tissue.
Pre-Lab Questions:
1.
Do you think the cancerous breast cells will be bigger, smaller, or the same as the healthy
breast cells?
2. Rewrite your answer to question one in the form of an If … Then … hypotheses.
EXERCISE 3: Normal and Cancerous Breast Tissue
Data Collection:
3. Select the mammary gland slide (Slide Cassette 4: TBA) from the slide loader. Using the 10X
objective, identify the cells and bring them into focus.
4. Carefully work your way through all the objectives focusing with each one until you reach the
60X objective and capture an image. Insert your image of normal breast tissue below.
5. Select the breast carcinoma slide (Slide Cassette 4: TBA) from the slide loader. Using the 10X
objective, identify the carcinoma cells and bring them in to focus.
6. Carefully work your way through all the objectives focusing with each one until you reach the
60X objective and capture an image. Insert your image of the breast carcinoma below.
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Analysis:
7. Using the method form exercise 1, measure and compare the size of the cells from the
mammary gland with breast carcinoma cells.
8. Are your results in correlation with what you have predicted earlier?
9. Rewrite your hypothesis to take into to account the new information you learned in this
exercise.
PRE-LAB EXERCISE 4: Normal and Cancerous Spleen
Leukemia is often thought of as a cancer of the blood which it is. However, leukemia is more accurately
described as cancer of the white blood cells. The white blood cells play a large role in the immune
response, which also involves many other tissues and organs. In this exercise, we are going to look at
the effect of leukemia on the spleen.
Pre-lab Questions:
1. What differences do you expect to see in the tissue samples taken from a normal spleen versus
a tissue sample taken from a leukemia infected spleen?
2. Write a hypothesis in the IF … THEN … format that predicts what effect you expect to see with
respect to fat and lumen size in an Atherosclerosis blood vessel.
EXERCISE 4: Normal and Cancerous Spleen
Data Collection:
3. Select the spleen slide from the slide loader. Using the 10X objective, identify the cells and bring
them into focus.
4. Carefully work your way through all the objectives, focusing with each one until you reach the
60X objective and capture an image. Insert your image of the normal spleen below.
5. Select the leukemia spleen slide from the slide loader. Using the 10X objective, identify the cells
and bring them into focus.
6. Carefully work your way through all the objectives focusing with each one until you reach the
60X objective and capture an image. Insert your image of the leukemia spleen below
Analysis:
7. Using the method from exercise 1, measure and compare the size of the cells in the normal
spleen versus the cells from the leukemia spleen.
8. What is the difference in the frequency of white blood cells in the normal spleen versus the
leukemia spleen?
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9. Are your results in correction with what you have predicted earlier?
10. Rewrite your hypothesis to take into account the new information you learned in this exercise.
ADD IF CARDIAC IS USED IN ADDITION TO OR AS A REPLACEMENT FOR SPLEEN
PRE-LAB EXERCISE 4A: Cardiac Muscle and Myocardial Infarction
One of the leading causes of death in the developed world is heart disease. A specific type of heart
disease is a Myocardial Infarction, commonly referred to as a heart attack, which is caused by the loss of
oxygen to part of the heart. It is often common for a heart attack to lead to the death of a small amount
of cardiac muscle which will be replaced by scare tissue.
Pre-Lab Questions:
1. What do you predict the differences between the normal cardiac muscle and the cardiac muscle
suffering from myocardial infarction will be?
2. Rewrite your answer to question #1 in the form of an If … Then … hypotheses.
EXERCISE 4A: Cardiac Muscle and Myocardial Infarction
Data Collection:
3. Select the cardiac muscle slide (Slide Cassette 4: TBA) from the slide loader. Using the 10X
objective, identify the cells and bring them into focus.
4. Carefully work your way through all the objectives focusing with each one until you reach the
60X objective and capture an image. Insert your image of cardiac muscle below.
5. Select the myocardial infarction slide (Slide Cassette 4: TBA) from the slide loader. Using the
10X objective, identify the cells and bring them into focus.
6. Carefully work your way through all objectives focusing with each one until you reach the 60X
objective and capture an image. Insert your image of the myocardial infarction muscle below.
Analysis:
7. Based on your observations, describe the difference you see between the two cardiac muscles
in the samples.
8. Are your results in correlation with what you have predicted earlier?
9. Rewrite your hypothesis to take into account the new information you learned in this exercise.
END IF CARDIAC IS USED IN ADDITION TO OR AS A REPLACEMENT FOR SPLEEN
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PRE-LAB EXERCISE 5: Cirrhosis of the Liver
At a first pass, the liver seems to be a relatively simple organ. Its appearance is basically uniform and
only interrupted by blood vessels. However, what the liver may lack in appearance it makes up for in
metabolic complexity. As we mentioned in the introduction, the liver is responsible for the metabolism
(removal) of excess fat. When this process is disrupted, fat will accumulate in the liver cells and lead to
problems and eventually disease. One thing that will lead to a disruption of fat metabolism is excessive
drinking (cirrhosis of the liver). In this lab, you will examine normal and diseased liver cells.
Pre-lab Questions:
1. What defects do you think you will see in the cells from the cirrhosis liver?
2. Write a hypothesis in the IF … THEN … format that predicts what effect you expect to see with
respect to a cirrhosis liver.
EXERCISE 5: Cirrhosis of the Liver
Data Collection:
1. Select the liver slide (Slide Cassette 4: TBA) from the slide loader. Using the 10X objective,
identify the cells and bring them into focus.
2. Carefully work your way through all objectives focusing with each one until you reach the 60X
objective and capture an image. Insert your image of the liver tissue below.
3. Select the cirrhosis slide (Slide Cassette 4: TBA) from the slide loader. Using the 10X objective,
identify the cells and bring them into focus.
4. Carefully work your way through all objectives focusing with each one until you reach the 60X
objective and capture an image. Insert your image of the cirrhotic liver tissue below.
Analysis:
7. Based on your observation, describe the difference you see between the two liver samples.
8. Are your results in correlation with what you have predicted earlier?
9. Rewrite your hypothesis to take into account the new information you learned in this exercise.
SUMMARY QUESTIONS:
1.
2.
3.
4.
5.
6.
7.
8.
9.
Explain the difference between etiology and pathogenesis.
Provide two examples of etiologic factors that may cause diseases in human.
Discuss the purpose of cellular adaptation in our body.
Provide an example of cellular adaptation and its cause(s).
Discuss the difference between reversible and irreversible cell injury and provide an example for
each.
What is the definition of anemia? List three major causes of anemia.
Explain the rationale for each of the following manifestations of anemia: pale skin, increased
heart rate, dizziness and fainting.
How does blood loss lead to anemia? Does all blood loss lead to anemia?
What is jaundice and why does it occur in some forms of anemia and not others?
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10. Why do red blood cells need iron?
11. Define leukemia?
12. What is the difference between acute and chronic leukemia? Discuss the predominant cell type
and the onset / course of the disease for each type of leukemia.
13. Discuss the role of vitamin K in blood coagulation?
14. What effect does liver disease have on coagulation?
15. Provide an example of a specific type of intracellular accumulation and its impact on the cell’s
metabolism and the overall health.
16. Discuss the role of diet in the development of atherosclerosis and its impact on the overall
function of the cardiovascular system.
17. IF CARDIAC IS USED: Describe the changes that occur in cardiac muscle that has suffered
myocardial infarction. How is the damaged tissue different than healthy cardiac tissue?
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PREPARING FOR THIS NANSLO LAB ACTIVITY:
Read and understand the information below before you proceed with the lab!
Scheduling an Appointment Using the NANSLO Scheduling System
Your instructor has reserved a block of time through the NANSLO Scheduling System for you to complete
this activity. For more information on how to set up a time to access this NANSLO lab activity, see
www.wiche.edu/nanslo/scheduling-software.
Students Accessing a NANSLO Lab Activity for the First Time
You must install software on your computer before accessing a NANSLO lab activity for the first time.
Use this link to access instructions on how to install this software based on the NANSLO lab listed below
that you will use to access your lab activity – www.wiche.edu/nanslo/lab-tutorials
1. NANSLO Colorado Node -- all Colorado colleges.
2. NANSLO Montana Node -- Great Falls College Montana State University, Flathead Valley
Community College, Lake Area Technical Institute, and Laramie County Community College.
3. NANSLO British Columbia Node -- Kodiak College.
Using the Microscope for a NANSLO Remote Web-based Science Lab Activity
We've provided you with three ways to learn how to use the microscope for this NANSLO lab activity:
1. Read these instructions.
2. Watch this short video https://www.youtube.com/watch?feature=player_embedded&v=m7w9ssIgVdw.
3. Print off these instructions to read (PDF version of the instructions.)
NOTE: The conference number in this video tutorial is an example. See “Communicating with
Your Lab Partners” below to determine the toll free number and pin to use for your NANSLO lab
activity.
MICROSCOPE RWSL LAB INTERFACE INSTRUCTIONS
The Remote Web-based Science Lab (RWSL) microscope is a high quality digital microscope located at
the NANSLO Node. Using a web interface as shown below, you can control every function of the
microscope just as if you were sitting in front of it.
The equipment control software shown below is written using the LabVIEW software from National
Instruments. The user interface is presented as a LabVIEW control panel which will be referred to as the
lab interface for the remainder of the document.
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Figure 1: Remote Web-based Science Lab (RWSL) Microscope Lab Interface
COMMUNICATING WITH YOUR LAB PARTNERS
As soon as you have accessed this lab interface, call into the toll free conference number shown on the
control panel to communicate with your lab partners and with the Lab Technicians. Use the PIN code
noted to join your lab partners. Only one person can be in control of the equipment at any one time so
talking together on a conference line helps to coordinate control of the equipment and creates a more
collaborative environment for you and your lab partners.
GAINING CONTROL OF THE MICROSCOPE
Right click anywhere in the grey area of the lab interface and choose “Request Control of VI” from the
dialogue box that appears when multiple students are using the microscope at the same time,. After
you request control, you may have to wait a short time before you actually receive control and are able
to use the features on this lab interface.
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Figure 2: Selecting "Request Control of VI"
RELEASING CONTROL OF THE MICROSCOPE
To release control of the microscope so that another student can use it, right click anywhere in the grey
area of the lab interface and choose "Release Control of VI" from the dialogue box that appears.
Figure 3: Selecting "Release Control of VI"
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MICROSCOPE CONTROLS
The Stage Controls allow you to adjust the visual of the specimen that has been placed on the stage of
the microscope, select lenses with various magnifications, and select whether or not the condenser lens
is in the light beam. Below are more specific instructions on using these controls. When using the
arrows on this lab interface, click and hold the arrow until the desired effect is achieved or click and wait
to view the result before clicking again. Quick clicks on the arrows may cause the system to lock up.
Figure 4: Microscope Controls - Stage, Objective & Condenser
Stage Controls: Using the left and right and up and down arrows found to the right of the microscope
image in the Stage Control area, moves the microscope stage which holds the specimen. These arrows
allow you to precisely control the position of the specimen on the stage.
1. Use the "Right" and "Left" arrows to move the Stage so that you can view the specimen from
left to right.
2. Use the "Backward" and "Forward" arrows to move the Stage so that you can view the top,
middle or bottom of the specimen.
3. Use the "Up" and "Down" arrows to move the stage closer or farther away from the objective
lens to bring a specimen into focus. BE CAREFUL! Don't move the stage too close to the lens.
When selecting the button between the "Up" and "Down" arrows, you can toggle between “Coarse” and
“Fine” focus. When the button is dark green and “Coarse/Fine” is displayed to the right of the button,
the microscope is in “Coarse” focus. When the button is bright green and “Fine” is displayed, the
microscope is in “Fine” focus. Typically, you will start with coarse focus which moves the stage in large
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increments and then use fine focus to complete your final focusing as it moves the stage in smaller
increments. There is no difference between the course and fine focus when using the 60X objective
NOTE: When you click on these arrows, the specimen appears to move in the opposite direction. Since
the objective stays fixed, the image moves in the opposite direction of the stage. This is how these
controls work on most microscopes so the "feel" of the microscope is preserved over the web.
Figure 5: Right/Left & Backward/Forward Stage
Controls
Figure
6: Up/Down Stage Controls & Coarse/Fine Focus
Control
Objective: A microscope mounts an objective lens very close to the object to be viewed. Depending on
need, different lenses with different power will be used on the microscope. This microscope feature
multiple objectives, each with different power, mounted on a rotating turret. The larger the
magnification numbers the greater the magnification. For example, if a specimen is viewed through a
40X objective lens, the magnifier in that lens displays the specimen 40 times larger than an equivalent
view as seen by the unaided eye. Remember that the ocular or other lenses also add to the
magnification.
This microscope has five lenses – 4X, 10X, 20X, 40X, and 60X. Use the arrows below the objective lens
box that indicates the magnification of the current objective lens to move to a higher or lower
magnification lens. If you have activated the “Picture-in-Picture” Preset 2 (see below) you will be able to
see the objective lens move when you select a new magnification.
Condenser: The condenser controls whether or not the condenser lens is in the light beam. You want to
have the condenser OUT for the 4x objective but IN for all the others.
SELECTING A CASSETTE AND LOADING SLIDES ONTO THE STAGE
There are two tabs on the lab interface. When you first access the lab interface, the "Microscope" tab is
displayed by default. Click on the Slide Loader tab at the top of the screen to access the controls for the
Slide Loader robot. There can be up to four cassettes available on the Slide Loader. These cassettes are
used to store slides, and each can hold up to 50 slides. The cassettes available to you are dependent on
the lab activity to be completed. Once a cassette has been selected, you will use the drop-down list to
select your slides.
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Figure 7: Select the Slide Loader Tab to select a cassette and slides.
EXAMPLE OF HOW TO LOAD SLIDES
In this example, we have selected Cassette #1. Using the drop-down menu, we have selected "1:
Colored Threads Whole Mount." Then, we selected the "Load" button. A message indicates that the
slide is loading. Using the picture-in-picture camera, you can watch this happening. The robotics selects
the slide and places it on the microscope stage.
Figure 8: Selecting the slide
"1: Colored Threads Whole Mount" from Cassette #1
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Notice that when a slide is actually on the microscope (or when it is being loaded or unloaded), the
cassette controls are greyed out so you cannot load a second slide until the first is removed. Once the
slide is on the microscope stage, it will be listed in the "Current Slide on Stage" box. The only thing that
the Slide Loader robot can do is return it to the cassette when the "Return Slide to Cassette" button is
selected.
Figure 9: "LOADING SLIDE ... PLEASE WAIT" is displayed
in the "Current Slide on Stage" window
Select the "Microscope" tab to perform the NANSLO lab activity. Once you are finished with the slide,
select the "Slide Loader" tab and select "Return Slide to Cassette" button. Once the slide is returned to
the cassette, the Slide Loader controls are again available to select another slide from the cassette.
ENHANCING THE MICROSCOPE IMAGE
The digital camera mounted on the microscope has a camera control unit that is equipped with a series
of image processing functions that enable you to quickly and easily correct imaging problems that arise
from low or high contrast, poor focus, insufficient or uneven illumination, sample shading or
discoloration and noise. The most common reason for uneven elimination is a light source that does not
completely fill the field of view on lower magnifications. The White Balance should be used only if the
image appears to be brown or gray, and you think you might need to adjust it (although it won't hurt
anything to click this button).
A choice of color modes can be selected in the Microscope Image area and are used to display the image
in different color palettes in order to highlight certain features. The default setting is "Normal."
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Figure 10: Microscope Image Special Effects and Other Image Controls for Camera
Here is a description of each option:
1. In the “Normal” mode, the sample is displayed in its true colors.
2. In the “Negative” mode, the sample is displayed in a color-inverted form, where red, green, and
blue values are converted into their complementary colors. The technique is useful in situations
when color inversion can be of benefit in exposing subtle details or in quantitative analysis of
samples.
3. In the “Blue Black” mode, the black portions of a grayscale negative sample are displayed in
blue. This mode is often useful to reveal details in samples having a high degree of contrast. The
“Blue Black” filter can aid you in examining a wide spectrum of difficult samples.
4. In the “Black & White” mode, a grayscale image of the sample is displayed.
5. In the “Sepia” mode, a brown scale (black and white) image of the sample is displayed. Although
typically this filter is of little utility, it can be employed to alter image color characteristics to
improve the visualization of sample detail.
6. At times, the sample may have an unacceptable color quality. Use “White Balance” calibration
to remove the color cast. This process is often referred to as white balancing.
7. Auto Exposure is on automatically. You do not need to do anything with Auto Exposure unless
you are adjusting the luminance. If you are doing so, you should turn off Auto Exposure by
clicking on the button. The green light is now off. Now adjust the luminance. See explanation
below.
Reference: http://www.microscopyu.com/articles/digitalimaging/dn100/correctingimages.html
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Auto Exposure is normally turned on, but you can turn it off if you want to play around with the
brightness of the light source and not have the microscope camera automatically adjust it. It is usually
best, though, to leave it turned on.
When you turn off the Auto Exposure, the button turns dark green. Some new controls appear that let
you turn the LED off or on, and also adjust the intensity of the light source. The intensity of the light
source can be increased or decreased manually with the dial that now appears next to the Objective
control when Auto Exposure is turned off.
Figure 11: Additional controls available when Auto Exposure is turned off
CAPTURING AND SAVING A MICROSCOPE IMAGE
When the “Capture Image” button is pressed, a high-resolution image of what is currently in the field of
view of the objective is captured. While the image is being captured, the button will be illuminated
bright green. The capture is complete when the light turns off. Be patient as this may take several
seconds to complete.
After the Capture Image light turns off, select the “View Captured Image” tab on the bottom of this
control panel to view the image.
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Figure 12: Click the capture image button (#1), wait till the green light goes off,
and then select the View Captured Image tab (#2)
After opening this image through the View Captured Image tab, you will need to take a snapshot of it
and save it to your computer. There are several ways to do this, depending on your operating system.
WINDOWS:
1.
Pressing the two keys ALT and Print Screen simultaneously will copy the active window into
your computer clipboard. Then you can past it into a document.
2. Windows 7 and above has a Snipping Tool program under Programs/Accessories which can
capture selected areas of the screen.
3. Right click on it and select "Copy" from the menu presented. After right clicking and selecting
Copy, just open a document and right click and select Paste. You can either paste it directly into
your lab report document or into another one for safe keeping until you use it later. You can
use drawing tools in your word processing editor to annotate this image so you can show your
instructor that you know what you were suppose to be looking for!
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Figure 13: Right click and select Copy to paste the image into a document.
MAC:
1. Press these three keys simultaneously –
. This will change your cursor icon into a
little cross.
2. Now press the spacebar, and the icon becomes a camera. Click in the image window you want
to take a snapshot of, and it will save the image to a file on your desktop.
There are lots of free screenshot utilities you can also use to capture this image.
If you are familiar with saving a document to your computer, you also can select “Save Image As” from
the pop-up menu, give the image a name and then select a location on your computer where you want
this image to be saved for future use.
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MICROSCOPE IMAGE VIEW WINDOW
The Image View Window displays the real-time video feed from the digital camera “looking through” the
microscope.
Figure 14: Image View Window
PICTURE-IN-PICTURE CONTROLS - CAMERA PRESET POSITIONS AND PAN-TILT-ZOOM CONTROLS
When you click on the "Picture-in-Picture" button, it turns bright green. A second real-time video feed
from another digital camera appears in the Image View Window. The controls shown in Figure 15 are all
operational when the Picture-in-Picture feature is selected.
Figure 15: Picture-in-Picture Image Controls
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CAMERA PRESETS
There are six camera preset positions.
Figure 16: Picture-in-picture Camera Preset 1 and 6
- Displays the microscope, microscope camera, and a
camera control unit projecting the sample on the
Stage.
Figure 17: Picture-in-picture Camera Preset 2:
Displays a closeup of the objective lens.
Figure 18: Picture-in-picture Camera Preset 3 Displays a closeup of the camera control unit
projecting the sample on the Stage.
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Figure 19: Picture-in-picture Camera Preset 4 Displays the microscope eye piece and
the camera mounted to the microscope.
Figure 20: Picture-in-picture Camera Preset 5 Displays the Condenser Lens underneath the Stage
that focuses the light on the sample. The
Condenser Lens controls the width of the beam. In
some instances you will want a tighter beam while
in other cases you will want a broader beam to
control the image quality. This setting has been
optimized for you.
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PAN, TILT, ZOOM CONTROLS FOR PICTURE-IN-PICTURE
For each camera preset view, additional camera options are available.
1. Use the up and down arrows to tilt the camera up or down.
2. Use the right and left arrows to pan right or left.
3. Use the left "Zoom OUT" arrow and right "Zoom IN" arrow to zoom out and in.
Figure 21: Picture-in-picture Camera - Example of "Zoom In" capability
For more information about NANSLO, visit www.wiche.edu/nanslo.
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