REMOTE LAB ACTIVITY
SUBJECT SEMESTER: ________________
TITLE OF LAB: Hematology
Lab format: This lab is a remote lab activity.
Relationship to theory (if appropriate): In this lab you will learn to identify the cellular components of
blood and how perturbations in its structure or composition can cause physiological problems.
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 - Mitosis and Meiosis Slide Set.
CONTENTS FOR THIS NANSLO LAB ACTIVITY:
Learning Objectives....................................................................................................................
Background Information ...........................................................................................................
Equipment .................................................................................................................................
Experimental Procedure ...........................................................................................................
Pre-lab Exercise 1: Microscopic Examination of the Formed Elements of Blood .....................
Exercise 1: Microscopic Examination of the Formed Elements of Blood .................................
Pre-lab Exercise 2: Normal Red Blood Cells and Sickled Cells Slide Observation ....................
Exercise 2: Normal Red Blood Cells and Sickled Cells Slide Observation .................................
Pre-lab Exercise 3: Normal Red Blood Cells and Thalassemia majors Slide Observation ........
Exercise 3: Normal Red Blood Cells and Thalassemia majors Slide Observation .....................
Pre-lab Exercise 4: Normal and Infectious Mononucleosis Blood Observation ......................
Exercise 4: Normal and Infectious Mononucleosis Blood Observation ..................................
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2-7
7
7
7
8-9
9
9
10
10
10
11
Exercise 5: ABO-RH Blood Typing ............................................................................................. 11 - 12
Summary Questions: Mitosis and meiosis experiment ............................................................ 13
Preparing to Use the Remote Web-based Science Lab (RWSL) ................................................ 14 - 28
LEARNING OBJECTIVES:
After completing this laboratory experiment, you should be able to do the following things:
1.
2.
3.
4.
5.
Know the function of each formed element in the blood.
Identify and distinguish between the different kinds of formed elements in the blood.
Measure the relative size of certain cells.
Identify, distinguish, and measure the size of healthy red blood cells and sickle-celled ones.
Identify, distinguish, and measure the size of healthy red blood cells and Thalassemia majors
ones.
6. Identify, distinguish, and measure the size of healthy white blood cells and infectious
mononucleosis ones.
7. Explain the basis for blood typing and matching for blood donation and identify appropriate
blood donors for a given recipient.
BACKGROUND INFORMATION:
Blood is the fluid that transports oxygen and nutrients to the cells and carries away carbon dioxide and
other waste products. Technically, blood is a transport liquid pumped by the heart (or an equivalent
structure) to all parts of the body after which it is returned to the heart to repeat the process. Blood is
the only tissue whose matrix is a complete fluid. It is a tissue, because it is a collection of similar
specialized cells that serve particular functions. These cells are suspended in a liquid matrix (plasma)
which makes the blood a fluid. If blood flow ceases, death will occur within minutes because of the
effects of an unfavorable environment on highly susceptible cells.
Blood cells known as formed elements are suspended in the plasma. Often clinical evaluations of the
blood characteristics of the patient are important in assessing the condition of the patient. Blood
characteristics vary according to age, sex, metabolic condition, health conditions, and genetic variations.
Plasma serves as a transport medium for delivering nutrients to the cells of the body and for
transporting waste products derived from cellular metabolism to the kidneys, liver, and lungs for
excretion. It is also a transport system for blood cells, and it plays a critical role in maintaining normal
blood pressure. Plasma helps to distribute heat throughout the body and to maintain homeostasis, or
biological stability, including acid-base balance in the blood and body.
Plasma is derived when all the blood cells—red blood cells (erythrocytes), white blood cells (leukocytes),
and platelets (thrombocytes)—are separated from whole blood. The remaining straw-colored fluid is
90–92 percent water, but it contains critical solutes necessary for sustaining health and life. Important
constituents include electrolytes such as sodium, potassium, chloride, bicarbonate, magnesium, and
calcium. In addition, there are trace amounts of other substances including amino acids, vitamins,
organic acids, pigments, and enzymes. Hormones such as insulin, corticosteroids, and thyroxin are
secreted into the blood by the endocrine system. Plasma concentrations of hormones must be carefully
regulated for good health.
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Figure 1: Blood is made up of multiple
components, including red blood cells, white
blood cells, platelets, and plasma.
Platelet: blood components". Art. Encyclopædia
Britannica Online. Web. 27 Apr. 2014.
http://www.britannica.com/EBchecked/media/
113906/Blood-is-made-up-of-multiplecomponents-including-red-blood
Red blood cells, also called erythrocytes, are the cellular component of blood. Millions of red blood
cells in the circulation of vertebrates give the blood its characteristic color and carry oxygen from the
lungs to the tissues. The mature human red blood cell is small, round, and biconcave. It appears
dumbbell-shaped in profile. The cell is flexible and assumes a bell shape as it passes through extremely
small blood vessels. It is covered with a membrane composed of lipids and proteins, lacks a nucleus,
and contains hemoglobin — a red, iron-rich protein that binds oxygen.
White blood cell, also called leukocytes, are cellular components of the blood that lack hemoglobin,
have nuclei, are capable of motility, and defend the body against infection and disease by ingesting
foreign materials and cellular debris, by destroying infectious agents and cancer cells, or by producing
antibodies (also called immunoglobulins, protective proteins produced by the immune system in
response to the presence of a foreign substance, called an antigen. Antibodies recognize and latch onto
antigens in order to remove them from the body.) A healthy adult human has between 4,500 and
11,000 white blood cells per cubic millimeter of blood. Fluctuations in white cell number occur during
the day. Lower values are obtained during rest and higher values during exercise. White cell count also
may increase in response to convulsions, strong emotional reactions, pain, pregnancy, labor, and certain
disease states, such as infections and intoxication.
Granulocytes, the most numerous of the white cells, rid the body of large pathogenic organisms such as
protozoans or helminths and are also key mediators of allergy and other forms of inflammation. These
cells contain many cytoplasmic granules, or secretory vesicles, that harbor potent chemicals important
in immune responses. They also have multilobed nuclei, and, because of this, they are often called
polymorphonuclear cells. On the basis of how their granules take up dye in the laboratory, granulocytes
are subdivided into three categories: neutrophils, eosinophils, and basophils.
The most numerous of the granulocytes—making up 50 to 80 percent of all white cells—are neutrophils.
They are often one of the first cell types to arrive at a site of infection where they engulf and destroy the
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infectious microorganisms through a process called phagocytosis. Eosinophils and basophils typically
arrive later. The granules of basophils contain a number of chemicals, including histamine and
leukotrienes, that are important in inducing allergic inflammatory responses.
Eosinophils destroy parasites and also help to modulate inflammatory responses.
Neutrophils are characterized histologically by their ability to be stained by neutral dyes and functionally
by their role in mediating immune responses against infectious microorganisms. Neutrophils move with
amoeboid motion.
Neutrophils are actively phagocytic. They engulf bacteria and other microorganisms and microscopic
particles. The granules of the neutrophil are microscopic packets of potent enzymes capable of digesting
many types of cellular materials.
An abnormally high number of neutrophils circulating in the blood is called neutrophilia. This condition
is typically associated with acute inflammation, though it may result from chronic myelogenous
leukemia, a cancer of the blood-forming tissues. An abnormally low number of neutrophils is called
neutropenia. This condition can be caused by various inherited disorders that affect the immune system
as well as by a number of acquired diseases, including certain disorders that arise from exposure to
harmful chemicals. Neutropenia significantly increases the risk of life-threatening bacterial infection.
Figure 2: Two neutrophils among many red
blood cells. The normal functions of
neutrophils are compromised in chronic
granulomatous disease. "Chronic
granulomatous disease". Photograph.
Encyclopædia Britannica Online. Web. 27
Apr. 2014.
http://www.britannica.com/EBchecked/me
dia/126495/Two-neutrophils-among-manyred-blood-cells
Eosinophils are characterized histologically by their ability to be stained by acidic dyes (e.g., eosin) and
functionally by their role in mediating certain types of allergic reactions. Eosinophils contain large
granules, and the nucleus exists as two non-segmented lobes.
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Figure 3: White blood cells in a field of red
cells. (Top left) Monocyte, (top centre)
basophil, (top right) platelets, (bottom left)
two neutrophils, (bottom right) lymphocyte
and eosinophil, respectively.
A. Owczarzak/Taurus Photos, Inc.,
"Eosinophil: white blood cells in field of red
cells". Photograph. Encyclopædia Britannica
Online. Web. 27 Apr. 2014.
http://www.britannica.com/EBchecked/medi
a/767/White-blood-cells-in-a-field-of-redcells-Monocyte
Basophils are characterized histologically by its ability to be stained by basic dyes and functionally by its
role in mediating hypersensitivity reactions of the immune system. Basophils are the least numerous of
the granulocytes and account for less than one percent of all white blood cells occurring in the human
body.
Agranulocytes do not contain any cytoplasmic granules or secretory vesicles. We distinguish two types:
lymphocytes and monocytes.
Lymphocytes, which are further divided into B and T cells, are responsible for the specific
recognition of foreign agents and their subsequent removal from the host.
o
o
B lymphocytes secrete antibodies.
Typically, T cells recognize virally infected or cancerous cells and destroy them, or they
serve as helper cells to assist the production of antibody by B cells.
Monocytes move from the blood to sites of infection where they differentiate further into
macrophages. These cells are scavengers that phagocytose whole or killed microorganisms and
are therefore effective at direct destruction of pathogens and cleanup of cellular debris from
sites of infection. Neutrophils and macrophages are the main phagocytic cells of the body, but
macrophages are much larger and longer-lived than neutrophils.
Platelets also called thrombocytes, are colorless, non-nucleated blood component that are important in
the formation of blood clots (coagulation). Platelets are found only in the blood of mammals.
Platelets are formed when cytoplasmic fragments of megakaryocytes, which are very large cells in the
bone marrow, pinch off into the circulation as they age. They are stored in the spleen. Some evidence
suggests platelets may also be produced or stored in the lungs where megakaryocytes are frequently
found.
Platelets play an important role in the formation of a blood clot by aggregating to block a cut blood
vessel and provide a surface on which strands of fibrin form an organized clot by contracting to pull the
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fibrin strands together to make the clot firm and permanent and, perhaps most important, by providing
or mediating a series of clotting factors necessary to the formation of the clot. Platelets also store and
transport several chemicals, including serotonin, epinephrine, histamine, and thromboxane. Upon
activation, these molecules are released and initiate local blood vessel constriction which facilitates clot
formation.
Figure 4: A micrograph of a round
aggregation of platelets (magnified
1000x).
Dr. F. Gilbert/Centers for Disease Control
and Prevention (CDC) (Image Number:
6645)
"Platelet". Photograph. Encyclopædia
Britannica Online. Web. 27 Apr. 2014.
http://www.britannica.com/EBchecked/
media/116860/A-micrograph-of-a-roundaggregation-of-platelets
Blood is a germ-free or axenic fluid; however, it is susceptible to infections by eukaryotic parasites,
bacteria or viruses, and some of these infections range from mild to fatal. For example:
Thalassemia is group of blood disorders characterized by a deficiency of hemoglobin, the blood
protein that transports oxygen to the tissues. Thalassemia (Greek: “sea blood”) is so called because
it was first discovered among peoples around the Mediterranean Sea among whom its incidence is
high.
In the mild form of the disease, Thalassemia minor, there is usually only slight or no anemia and life
expectancy is normal. Occasionally, complications occur involving slight enlargement of the spleen.
Thalassemia major is characterized by severe anemia, enlargement of the spleen, and body
deformities associated with expansion of the bone marrow.
Mononucleosis is a viral infection causing fever, sore throat, and swollen lymph glands, especially in
the neck. Mononucleosis, or mono, is often spread by saliva and close contact. It is known as "the
kissing disease" and occurs most often in those ages 15 to 17. However, the infection may develop
at any age. Mono is usually linked to the Epstein-Barr virus (EBV), but can also be caused by other
organisms such as cytomegalovirus (CMV). Mono may begin slowly with fatigue, a general ill feeling,
headache, and sore throat. The sore throat slowly gets worse. Your tonsils become swollen and
develop a whitish-yellow covering. The lymph nodes in the neck are frequently swollen and painful.
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References:
1. "blood." Encyclopedia Britannica. David H. Yawn, M.D. Last updated 6/27/2013.
http://www.britannica.com/EBchecked/topic/463483/plasma
2. "plasma." Encyclopedia Brintannica. David H. Yawn, M.D. Last updated 6/27/2013.
http://www.britannica.com/EBchecked/topic/463483/plasma
3. MedlinePlus, US Library of Medicine.
http://www.nlm.nih.gov/medlineplus/ency/article/000591.htm
4. "thalassemia." Encyclopedia Britannica. The Editor of the Encyclopedia Britannica. Last.
Updated 10/27/2013. http://www.britannica.com/EBchecked/topic/589769/thalassemia
EQUIPMENT:




Paper
Pencil/pen
Slides
o Human blood smear
o Human sickle cell anemia
o Thalassemia majors smear
o Infectious mononucleosis
Computer with Internet access (for the 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: Microscopic Examination of the Formed Elements of Blood
In this exercise you will:




Use the microscope to distinguish between the different types of formed elements in blood.
Identify the red blood cells from different white blood on images you have taken.
Measure and compare the relative size of red blood cells and the monocytes.
Compare the nucleus shape of the different white blood cells.
Pre-Lab Questions:
1.
Using what you know about the formed elements in blood, do you think the size and shape of
the red blood cells will be bigger, smaller, or the same as the ones of the monocytes?
2. Rewrite your answer to question #1 in the form of an If . . . Then . . . hypothesis.
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EXERCISE 1: Microscopic Examination of the Formed Elements of Blood
Data Collection:
3. Select the human blood smear slide (Slide Cassette 3: #21) from the slide loader. Using the 10X
objective, identify the blood cells and bring them in to focus.
4. Carefully working your way through all the objectives focusing with each one until you reach the
60X objective, capture an image. Insert your images of normal red blood cells, neutrophils,
eosinophils, basophils, lymphocytes and monocytes below.
5. Label the three different parts of a cell (nucleus, cytoplasm and cell membrane) and the
granules of the white blood cells.
6. Describe the shape of the red blood cells and the different white blood cells and their nuclei.
7. Were you able to see the nucleus of the red blood cells? Explain your answer.
8. Next we are going to measure the size of the red blood cells and the monocytes. 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 is 205µm.
9. Now if we use the image in figure 1 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).
Figure 5: Measurements
10. Now if we divide 37mm/136mm = 0.272 which we multiply the total length of the field of view
by so 0.272 * 205µm = 55.77 µm rounded for significant figures gives us a cell size of 56µm.
11. Measure the difference in the size of the red blood cells and compare that to the size of the
monocytes.
12. Are your results in correlation with what you have predicted earlier?
13. Rewrite your hypothesis in light of our new information you collected in this exercise.
Analysis:
14. Describe the function of each cell type in the table blow.
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Cell Type
Erythrocytes
Neutrophils
Eosinophils
Basophils
Monocytes
Platelets
Function
PRE-LAB EXERCISE 2: Normal Red Blood Cells and Sickled Cells Slide
Observations
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 2 Questions:
1. Do you predict the size of normal red blood cells to be smaller, bigger, or the same as the sickled
celled ones?
2. Rewrite your answer to question one in the form of an If … Than … hypothesis.
EXERCISE 2: Normal Red Blood Cells and Sickled Cells Slide Observations
Data Collection:
3. Select the Sickle Cell Anemia slide (Slide Cassette 3: #22) from the slide loader. Using the 10X
objective, identify the blood cells and bring them in to focus.
4. Carefully working your way through all the objectives focusing with each one until you reach the
60X objective, capture an image. Insert your picture of sickled cells below.
Analysis:
5. Utilizing your normal blood image form exercise 1 and the method from exercise 1 determine
the length of the normal and sickle red blood cells.
6. Based on your observation, describe the difference between the shape of normal red blood cells
versus sickled cells.
7. Are your results in correlation with what you have predicted earlier?
8. Rewrite your hypothesis to take into account the new information you have learned in this
exercise.
9. What is the impact of sickle cell anemia on oxygen transport?
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PRE-LAB EXERCISE 3: Normal Red Blood Cells and Thalassemia majors Slide
Observations
In this lab exercise you will measure and compare the relative size and shape of normal red blood cells
with Thalassemia major ones.
Pre-lab 3 Questions:
1. Do you predict the size of normal red blood cells to be smaller, bigger or the same as
Thalassemia majors ones?
2. Rewrite your answer to question one in the form of an If … Than … hypothesis.
EXERCISE 3: Normal Red Blood Cells and Thalassemia majors Slide Observations
Data Collection:
3. Select the Thalassemia majors slide (Slide Cassette 3: #32) from the slide loader. Using the 10X
objective, identify the blood cells and bring them in to focus.
4. Carefully working your way through all the objectives focusing with each one until you reach the
60X objective, capture an image. Insert your picture of Thalassemia major cells below.
Analysis:
5. Utilizing your normal blood image form exercise 1 and the method from exercise 1 to determine
the diameter of the normal red blood cells and the Thalassemia majors ones.
6. Based on your observation, describe the difference between the size of normal red blood cells
versus the Thalassemia majors cells.
7. Are your results in correlation with what you have predicted earlier?
8. Rewrite your hypothesis to take into account the new information you have learned in this
exercise.
9. What is the impact of large pale red blood cells on the blood circulation and the overall body
health? Explain your answer.
PRE-LAB EXERCISE 4: Normal and Infectious Mononucleosis Blood Observations
In this lab exercise you will measure and compare the relative size and shape of normal white blood cells
with infectious mononucleosis ones.
Pre-lab 4 Questions:
1. Do you predict the size of normal white blood cells to be smaller, bigger or the same as the
infectious monocudeosis ones?
2. Rewrite your answer to question one in the form of an If … Than … hypothesis.
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EXERCISE 4: Normal and Infectious Mononucleosis Blood Observations
Data Collection:
Select the infectious mononucleosis slide (Slide Cassette 3: #25) from the slide loader. Using
the 10X objective, identify the blood cells and bring them in to focus.
3. Carefully working your way through all the objectives focusing with each one until you reach the
60X objective, capture an image. Insert your picture of infected white blood cells below.
Analysis:
5. Utilizing your normal blood image form exercise 1 and the method from exercise 1, determine
the diameter of the normal white blood cells and the infectious mononucleosis ones.
6. Based on your observation, describe the difference between the size of these cells.
7. Are your results in correlation with what you have predicted earlier?
8. Rewrite your hypothesis to take into account the new information you have learned in this
exercise.
9. What is the impact of the infectious mononucleosis on the overall function of the immune
system? Explain your answer.
EXERCISE 5: ABO-RH Blood Typing
Blood typing is the classification of blood in terms of distinctive inherited characteristics that are
associated with the antigens located on the surface of red blood cells (erythrocytes). The ABO and the
Rh blood groups are among those most commonly considered. Identification of these determinants has
become indispensable in connection with blood transfusion, because the recipient and donor must have
the same, or compatible, blood groups. Otherwise, hemolysis (destruction) or coagulation (clotting)
results from interaction of an antigen on the red blood cells of one with an antibody in the serum of the
other.
ABO blood group system is the classification of human blood based on the inherited properties of red
blood cells (erythrocytes) as determined by the presence or absence of the antigens A and B which are
carried on the surface of the red cells. Persons may thus have type A, type B, type O, or type AB blood.
The A, B, and O blood groups were first identified by Austrian immunologist Karl Landsteiner in 1901.
Blood containing red cells with type A antigen on their surface has in its serum (fluid) antibodies against
type B red cells. If, in transfusion, type B blood is injected into persons with type A blood, the red cells in
the injected blood will be destroyed by the antibodies in the recipient’s blood. In the same way, type A
red cells will be destroyed by anti-A antibodies in type B blood. Type O blood can be injected into
persons with type A, B, or O blood unless there is incompatibility with respect to some other blood
group system also present.
To perform a blood typing test, anti-A and anti-B sera are each separately mixed with a drop of sample
blood and observed for agglutination (or clumping).
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Rh blood group system is a system for classifying blood groups according to the presence or absence of
the Rh antigen, often called the Rh factor, on the cell membranes of the red blood cells (erythrocytes).
The designation Rh is derived from the use of the blood of rhesus monkeys in the basic test for
determining the presence of the Rh antigen in human blood. The Rh blood group system was discovered
in 1940 by Karl Landsteiner and A.S. Weiner. Since that time, a number of distinct Rh antigens have been
identified, but the first and most common one, called RhD, causes the most severe immune reaction and
is the primary determinant of the Rh trait.
The Rh antigen poses a danger for the Rh-negative person, who lacks the antigen, if Rh-positive blood is
given in transfusion. Adverse effects may not occur the first time Rh-incompatible blood is given, but the
immune system responds to the foreign Rh antigen by producing anti-Rh antibodies. If Rh-positive blood
is given again after the antibodies form, they will attack the foreign red blood cells, causing them to
clump together, or agglutinate. The resulting hemolysis, or destruction of the red blood cells, causes
serious illness and sometimes death.
A similar hazard exists during pregnancy for the Rh-positive offspring of Rh-incompatible parents when
the mother is Rh-negative and the father is Rh-positive. The first child of such parents is usually in no
danger unless the mother has acquired anti-Rh antibodies by virtue of incompatible blood transfusion.
During labor, however, a small amount of the fetus’s blood may enter the mother’s bloodstream. The
mother will then produce anti-Rh antibodies which will attack any Rh-incompatible fetus in subsequent
pregnancies. This process produces erythroblastosis fetalis, or hemolytic disease of the newborn, which
can be fatal to the fetus or to the infant shortly after birth. Treatment of erythroblastosis fetalis usually
entails one or more exchange transfusions. The disease can be avoided by vaccinating the mother with
Rh immunoglobulin after delivery of her firstborn if there is Rh-incompatibility. The Rh vaccine destroys
any fetal blood cells before the mother’s immune system can develop antibodies.
Reference: http://www.britannica.com/EBchecked/topic/69880/blood-typing
In the table below, predict the agglutination reactions for each of the following blood types.
Blood Type
A+
A–
B+
B–
AB +
AB –
O+
O–
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Agglutination Reaction (Clumping – yes or no)
Anti-A Serum
Anti-B Serum
Anti-Rh Serum
Last Updated May 1, 2014
SUMMARY QUESTIONS:
Answer the following questions.
3. Which of the formed elements are referred to collectively as granulocytes?
4. Which of the formed elements are referred to collectively as agranulocytes?
5. When a patient is admitted to the hospital, one of the first procedures that health professionals
perform is a blood draw. One lab value that is checked is a white blood cell (WBC) count. If the
WBC count is elevated, this could indicate the presence of inflammation. Further analysis (called
a differential) is performed to determine the relative prevalence of different types of white
blood cells and the potential cause of the inflammation. What might be causing the
inflammation if (use your textbook for reference:)
a. Neutrophils were elevated? Explain your reasoning.
b. Lymphocytes were elevated? Explain your reasoning.
c. Eosinophils were elevated? Explain your reasoning.
6. If someone is diagnosed with "microcytic, hypochronic anemia," how would you describe the
hemoglobin content and size of his or her red blood cells?
7. Iron is necessary to produce hemoglobin. In iron-deficiency anemia, would you expect the red
blood cells to be hyperchronic, normochronic, or hypochronic? Explain.
8. What is the difference between antigen and antibody?
9. If your blood type is A-, which antigens are present on your red blood cells? What if your blood
type is AB+?
10. With the ABO blood types, individuals will have circulating antibodies to antigens that are not
present in their own blood. Identify the antibodies (A, B, both A and B, or none) produced by
the following individuals:
a.
b.
c.
d.
An individual with type A blood:
An individual with type B blood:
An individual with type O blood:
An individual with type AB blood:
11. Based on your results for each patient, identify the blood types he/she could receive using the
following choices. Note: Some patients may be able to receive several types of blood.
A+
A
B+
B
AB+
AB
O+
O
Patient 1 could receive _____________________________
Patient 2 could receive _____________________________
Patient 3 could receive _____________________________
12. Patient 4 could receive _____________________________
13. What is a universal donor? Which of these patients would be considered universal donor?
14. What would happen to a type O patient if he receives type A or B blood?
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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 grayed 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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