hematology “the study of blood” microscope...

Name: _________________ Date: _________________ BIO 30S Circulatory System

Hematology “The Study of Blood” Microscope Lab

IntroductionThe average male has about 5L of blood and women have 15% less. 1/4 of the total weight of the human body is made up of blood. About 45% of the blood volume is composed of specialized cells (erythrocytes, leukocytes and thrombocytes). These cells are suspended in an amber fluid called PLASMA. One drop of blood will contain as many as 250 000 000 red blood cells and 400 000 white blood cells.

Blood has 3 main functions:1. Carries to all living cells the nutrients needed for growth, maintenance and repair2. Transports oxygen and carbon dioxide3. Destruction of invading organisms

Purpose• To observe red and white blood cells under a microscope

Materials• Microscope• Prepared blood smear

Procedure AStudy a prepared slide of a blood smear. Note the disc-shaped red blood cells and the larger, less numerous white blood cells. Use the slide to complete the chart below and answer the following questions.

a) Based on your observations, complete the chart below that compares and contrasts red blood cells and white blood cells

b) Estimate how many red blood cells would fit across the field of high power lens.c) Using a value of 500 um, show your work and calculations to calculate the diameter of a single red blood cell.d) Explain why the centres of red blood cells are much lighter in colour.e) What is the purpose of the stain?

Procedure BUse the following information and diagrams to answer the questions that follow

Neutrophil• most common type of white blood cell• chemically attracted to any foreign matter that enters your body• while searching for foreign matter(such as bacteria and viruses) it can move like an amoeba (can move by throwing

out a jelly like arm and pouring the rest of its body toward it)• can leave the blood and circulatory system by squeezing through the pores of small blood vessels• their nuclei have 3 or more lobes, connected to one another by thin strands

CATEGORY RED BLOOD CELL WHITE BLOOD CELL

Relative size

Presence of nucleus?

Colour

Functions

Shape

Relative number, amount


Monocytes• destroy foreign invaders by a process called phagocytosis in which the leukocyte surrounds a foreign invader with

its jelly like body and digests it• has a large nucleus and can reach a diameter of 20 microns

Lymphocytes• lymphocytes are unusual because they can change into other types of blood cells• they can change into erythrocytes, monocytes and other types of white blood cells• look similar to monocytes (both have large nuclei) but a monocyte is much larger• while monocytes may be 20 microns in diameter, a lymphocyte is only about 8 microns in diameter (about the size

of an erythrocyte)

Basophils• release a chemical called heparin which appears to prevent blood from clotting in the arteries and veins• are about 12 microns in diameter and have a bilobed nucleus. The cytoplasm of the basophil is filled with large,

irregularly shaped granules

Eosinophils• believed to help you to stay healthy by removing poisonous substances from your tissues• are easy to identify because their nuclei are bi-lobed and look like stereo headphones

Analysis1. For each of the 5 types of white blood cells include the following: a) name b) function c) diagram

2. Neutrophils and monocytes can move like amoebas. Describe this kind of movement.

3. Neutrophils and monocytes can eat foreign invaders by a process called phagocytosis. Describe this process.

4. The number of leukocytes in the blood increase after a heavy meal. Why do you think this happens?

5. List the name of the leukocyte(s) that has the following properties:

a) able to prevent blood from clotting

b) absorbs poisonous substances from the tissues

c) phagocyte (2)

d) releases heparin

A B C D E


Blood Analysis Problems (Hematocrit)

1. Determine the normal hematocrit by using the following formula: Hematocrit = red blood cell volume x 100 total blood volume

a) Calculate and record the hematocrit of the normal subject.b) Calculate and record the hematocrit of person A, B, C and D.

2. A device called a hemacytomater is used to measure the amount of hemoglobin present. Red blood cells have the ability to concentrate hemoglobin to about 34 g/100mL of blood. Readings below 15 g/100mL of blood indicate anemia. Blood appears pale if hemoglobin levels are low.

c) Which subject (A,B, C or D) has a low level of hemoglobin? Explain.

3. Cancer of the white blood cells is called leukemia. Like other cancers, leukaemia is associated with rapid and uncontrolled cell production.

d) Which subject (A,B, C or D) might be suffering from leukemia? Explain.

4. Although heatocrits produce some information about blood disorders, most physicians would not diagnose leukaemia on the basis of one test.

e) What other conditions might explain the hematocrit reading you chose for answer d? Explain?

5. Lead poisoning can cause bone marrow destruction.f) Which of the subjects might have lead poisoning? Explain.g) Which subject lives at a high altitude? Explain.

6. Recently, athletes have begun to take advantage of the benefits of extra red blood cells. 2 weeks prior to a competition, a blood sample is taken and centrifuged and the red blood cell component is stored. A few days before the event, the red blood cells are injected into the athlete.

h) Why would athletes remove blood cells only to return them to their body later?

7. A physician notes fewer red blood cells and prolonged blood clotting times in a patient. White blood cell numbers appear to have increased, but further examination reveals that only the granulocyte numbers have increased, while the a granulocytes have decreased. In an attempt to identify the cause of the anomaly, the physician begins testing the bone marrow.

i) Why did the physician suspect the bone marrow?j) Predict what might have caused the problem.

8. Individuals who work in a chemical plant are found to have unusually high numbers of leukocytes. A physician calls for further testing.

k) Hypothesize about the physician’s reasons for concern.l) Why might the physician check both bone marrow and lymph node areas of the body?


Blood Typing (Part A)

1. The following illustrates how scientists determine blood type by cross matching. Heparin, an anticoagulant, is added to both a known and unknown blood sample to prevent blood clotting. The samples are then placed in a centrifuge and separated into components. The red blood cells from the known sample, Type A, are mixed with serum of the unknown sample. (NOTE: SERUM is plasma without the clotting factor).

In the next step, the red blood cells from the unknown sample are mixed with the serum from Type A blood. The data are provided int he following diagram.

PREDICT THE BLOOD TYPE OF THE UNKNOWN SAMPLE. EXPLAIN YOUR REASONING.

2. The serum containing antibody A will cause agglutination of donor blood that contains antigen A. The serum containing antibody B will agglutinate donor blood that has antigen B.

USE THE DATA BELOW TO PREDICT THE BLOOD TYPE OF EACH SAMPLE.

3. What would happen is blood Type A was transfused into people with:a) blood type Bb) blood type Ac) blood type Od) blood type ABProvide an explanation for each case.

4. How does Rh+ blood differ from Rh- blood?5. Explain why erythroblastosis fettles may affect a woman’s second and third child, but not her first.


Blood Typing (Part B)

Blood typing is helpful for determining the origin of a blood sample at a crime scene, helping to determine paternity without a DNA profile analysis.

MethodTest the blood samples at Stations 1-4.You may do this in groups.

Scenario 1The parents of baby Smith are concerned that there may have been a switch of their child at birth. Baby Smith had an accident and they found that the blood type of baby Smith is AB. The blood types of Mr. and Mrs. Smith are thought to be both B. The blood types of the parents need to be determined to see if there is any validity to their concern.

a) Sample 1 - Mrs. Smith Results of Testing ______________

b) Sample 2 - Mr. Smith Results of Testing ______________

c) What are your conclusions from your analysis?

Scenario 2A crime scene happened in room 23 in early November. You may have heard the CSI team talking about it and analyzing data/forensic evidence and DNA in the lab. It is your job to analyze 2 blood samples taken from the two suspects to see if they match anything at the crime scene. At the crime scene, B type blood was found on the pencil sharpener and on Miss Hagman’s scissors.

d) Sample 3 - Suspect A Results of Testing _______________

e) Sample 4 - Suspect B Results of Testing _______________

f) What are your conclusions from your analysis?


Investigating Pulse RateBackground

The heart pumps through blood vessels to all parts of the body. With each contraction of the heart, blood is forced into the arteries. This surge of pressure is felt in the arteries as the PULSE. The rhythmic pulse can be felt at any place where an artery is close to the surface of the body and can be pressed against some firm tissue.

The pulse rate is exactly equal to the heartbeat rate. Medical personnel use the pulse rate as one indication of how the heart is functioning. Heart rate is influenced by many things, such as age, sex, physiological state, psychological state, and temperature. In this activity, you will investigate how several of these factors influence the pulse rate.

PurposeIn this activity you will…

1. Feel a pulse and determine pulse rates.2. Determine the effect on pulse rate of standing at attention, holding your breath, deep breathing and

exercise.3. Make a line graph to show the effect of exercise on pulse rates.

MaterialsClock/Watch

Procedure1. Work with a partner. Throughout this experiment, you and your partner will take turns being the subject and the

experimenter. First you must learn how to take a pulse. Study Figure 1 to see how to locate the pulse in your partner’s wrist.

Table 1: Determining Resting Pulse Rate

2. After you have sat quietly for 1 minute, have your partner count your pulse rate for 15 seconds.a) Record this number (your pulse rate for 1 sec) in Table 1.b) Determine your pulse rate for 1 minute by multiplying your answer in “a” by 4. Record this number (your

pulse rate per minute) in Table 1.

3. Repeat Step 2, two more times. Then switch roles with your partnerc) Record your average resting pulse rate (Trial 1 + Trial 2 + Trial 3) and record it in Table 1, and also in

Table 2 for 3 your gender (on the

board).

4. The subject should stand stiffly at attention for 2 minutes. Then, while the subject is still standing at attention, the pulse should be taken by the experimenter for 15 seconds. Switch roles.d) Determine your pulse rate by multiplying this number by 4. Record your AT ATTENTION pulse rate in Table 2.

5. While seated, the subject should take a deep breath, exhale part of it, and hold the breath as long as possible. While the breath is being held, the subject’s pulse should be taken by the experimenter for 15 seconds. Then switch roles.

TRIAL PULSE RATE/15 SEC PULSE RATE/MIN

1

2

3

Average =


e) Determine your pulse rate by multiplying this number by 4. Record your BREATH-HOLDING pulse rate in Table 2.

6. While seated, the subject should take deep breaths regularly for 30 seconds. After the first 15 seconds, the pulse of the subject should be taken by the experimenter for the remaining 15 seconds of deep breathing. Then switch roles.f) Determine your pulse rate by multiplying this number by 4. Record your DEEP-BREATHING pulse rate in Table 2.

7. The time needed for your pulse to return to the sitting pulse rate is called RECOVERY TIME. The subject should step up and down from a sturdy chair, run in place or do jumping jacks for 1 minute. Immediately after exercise, the subject should sit and the pulse should be taken for 15 seconds. Then it should be taken again after 45 seconds, so that a 15 second pulse rate is taken EVERY MINUTE FOR 6 MINUTES. Switch roles.g) Determine your pulse rate by multiplying this number by 4. Record your AFTER-EXERCISE pulse rate in Table 2.

8. Assuming that your stroke volume is 70mL/beat, calculate your cardiac output for each of the activities using the following formula:

CARDIAC OUTPUT = STROKE VOLUME (mL/beat) x HEARTBEAT (beats/min)

h) Record your CARDIAC OUTPUT for each activity in Table 2.

Table 2: Effect of Activity on Pulse Rate

Analysis1. Copy the table of male and female resting rates/min from your class.a) Calculate the average resting rate for males and females. SHOW YOUR WORK.b) Explain how your resting pulse rate compares with the average rate for your gender.c) Compare the average male and female pulse rates for the resting pulse rates.

2. Explain how holding your breath affects your pulse rate.3. Which activity increased your pulse rate the most. EXPLAIN WHY.4. Why do athletes often have a lower pulse rate than non-athletes?5. Make a line graph to show what happens to your pulse rate after exercise. Put “TIME (in minutes)” on the

horizontal axis and “PULSE (per minute)” on the vertical axis. PLEASE USE GRAPH PAPER!

ACTIVITY PULSE RATE/15 SEC PULSE RATE/MIN CARDIAC OUTPUT (mL/min)

At-Attention

Breath-Holding

Deep-Breathing

After exercise (0 min)








Measuring Blood PressurePurposeUse a sphygmomanometer and stethoscope to measure systolic and diastolic blood pressure.

Background to Activity AThe survival of any organism depends on its ability to establish an internal environment that will keep individual cells alive and healthy. The maintenance of this internal environment in a steady state is called homeostasis. In complex organisms such as humans, homeostasis can only be maintained with a transport system that meets a wide range of needs. The blood, heart, and circulatory vessels carry out the necessary transport function. Contraction of the ventricles of the heart forces blood into the arteries and causes an increase in blood pressure. As the ventricles relax, blood pressure drops. As a result, blood pressure cycles between a high and a low. The highest pressure reached in the cycle is called the systolic pressure and the lowest pressure reached is the diastolic pressuree. Blood pressure is expressed as the height in millimetres that it will raise a column of mercury (mm Hg). The systolic pressure is written first and the diastolic pressure second (ex. 120/80 mm Hg). Baroreceptors located in the carotid arteries and aortic arch constantly monitor blood pressure and send nerve impulses to the brain. The brain sends nerve impulses to the heart, arterioles, and other organs to increase or decrease the blood pressure as needed. It is standard medical procedure to take blood pressure readings in the brachial artery of the arm, at the level of the heart. Blood pressure is routinely measured with a sphygmomanometer (Figure 1). The sphygmomanometer consists of an inflatable cuff, a pump, a gauge graduated in millimetres of mercury, and an exhaust valve with a screw control. The cuff is wrapped around the upper arm just above the elbow below the biceps. When the pressure in the cuff exceeds that in the artery, the artery collapses and blood flow stops. The pressure in the cuff is allowed to fall gradually by opening the exhaust valve. As the pressure in the cuff drops, it reaches a point at which the pressure of the blood forces the artery open slightly, allowing a turbulent flow of blood to pass. The turbulence sets up vibrations in the artery that are heard as sounds in the stethoscope (called Korotkov sounds). When the sound first becomes audible, it is a sharp thumping. The cuff pressure at which the sound is first heard is read as the systolic blood pressure. As pressure in the cuff decreases, the sharp thumping sound becomes louder and then muffles. The cuff pressure at which the sound disappears is read as the diastolic pressure.

Materials• Sphygmomanometer• Stethoscope• Alcohol swabs• Timer

IntroductionFor this activity, you will work in groups of 4 and will take turns measuring each other’s blood pressure using a sphygmomanometer and stethoscope. One of you will serve as the test subject, one as the examiner, one as the data recorder, and one as the timer. Then you will switch roles and repeat the activity.Note: These lab results are determined for experimental purposes only. They are not a substitue for regular, professional health care and diagnosis.

ProcedureTiming is important, so read the instructions before you begin the activity. The test subject should be seated, with sleeves (if any) rolled up.

The experimenter should:• Clean the ear pieces of the stethoscope with an alcohol swab before and after use.• Never leave an inflated cuff on anyone’s arm for more than a few seconds.

1. Inspect the sphygmomanometer. Be certain that the exhaust valve is open and that the cuff is completely deflated.

2. Wrap the cuff snugly, but not tightly, around the upper arm 2 to 3 cm above the bend in the elbow.3. Place the bell of the stethoscope directly below the cuff in the bend of the elbow.4. Close the exhaust valve of the bulb (pump) and rapidly inflate the cuff by squeezing the bulb until the pressure

gauge goes past 200 mm Hg.5. Open the exhaust valve just enough to allow the pressure to drop slowly, by about 2-5 mm Hg/sec.6. As the pressure falls, listen with the stethoscope for the first appearance of a clear thumping or tapping sound.

The pressure at which you first hear this sound in the systolic pressure. Record systolic pressure in Table 1.


7. Continue to listen as the pressure falls. The sound will become muffled and then louder. When the sound disappears, note the pressure. Record this measurement in Table 1 as the diastolic pressure.

8. Open the exhaust valve to completely deflate the cuff. Allow the subject to relax for 30 to 60 seconds before proceeding.

9. Repeat steps 1 through 8 two more times, to complete trials 2 and 3. Determine the subject’s average systolic and diastolic pressures.

Table 1: Blood Pressure While Seated

Background to Activity BPhysical fitness involves many components and can be defined in many ways (a champion gymnast, for example, might perform poorly in a marathon). The following tests are chosen to determine the ability of your cardiovascular system to adapt to change. This is one measure of general physical fitness. As you proceed, be alert to signs of dizziness or faintness in the test subject and be ready to steady or catch the subject if you are needed.

**Notify your teacher of any medical condition that might make it inadvisable for you to participate in any of these tests.**

Procedure1. The subject should recline for 5 minutes. After 5 minutes, take the subject’s systolic pressure and record it in

Table 2.2. The subject should remain reclining for 2 minutes after Step 1 and then stand up with arms down at the sides.

Immediately take the systolic pressure and record the data in Table 2.

**Caution: It is possible to become dizzy after standing in this manner. If the test subject becomes unsteady, becomes pale, or complains of feeling faint, seat them at once. Instruct them to lower their head between their knees and keep it down until the sensation passes.**

3. Determine the change in systolic pressure by subtracting the reclining systolic pressure from the standing systolic pressure. Record this data in Table 2.

Systolic Diastolic

Trial 1

Trial 2

Trial 3

Average


Table 2: Change in Systolic Pressure from Reclining to Standing

Calculations and AnalysisThe adaptability of the heart can be observed during exercise, when the metabolic activity of skeletal muscles increases. The cardiovascular system, consisting of the heart and blood vessels, responds to exercise with an increase in heart rate and strength of contraction with each beat, resulting in a higher cardiac output (cardiac output = quantity of blood pumped through the heart per unit of time) and blood pressure. Positive pressure is created by forceful contraction of the left ventricle of the heart, measured as systole. It is maintained during relaxation of the ventricle by closure of the aortic valve and recoil of arteries, measured as diastole ( see Figure 3). Mean arterial pressure (MAP) is a useful measure of the amount of oxygen getting to the cells, and is not a simple average of systolic and diastolic blood pressures. This is because diastole continues for twice as long as systole. MAP can be reasonably approximated using the equation:

systole + 2 diastole = MAP3

Calculate your MAP using your average systole and diastole values.

My MAP is ____________.

SYSTOLIC (mmHg)

Reclining Systolic Pressure

Standing Systolic Pressure

Change in Systolic Pressure

Change (mmHg) Points

Rise of 8 or more 3

Rise of 2-7 2

No rise 1

Fall of 2-5 0

Fall of 6 or more -1


Questions1. Why is it important that the subject’s arm be at heart level when taking blood pressure measurements?

2. Describe what happens to the blood in the artery of the arm as the pressure changes in the cuff during a

blood pressure reading.

3. The blood pressure readings correlate with different stages in the heartbeat. Explain which heartbeat

stage correlates with which blood pressure reading.

4. Consider two large mammals: a giraffe and a rhinoceros. If both animals were standing and relaxed,

which would you expect to have the higher blood pressure? Explain your answer.

5. Explain what would happen to your blood pressure right when you finished exercising strenuously. And

15 minutes later?

6. Do you think a higher or lower MAP would be healthy? Why?


Lung CapacityPurposeTo measure your tidal volume, expiratory reserve and vital capacity.

Materials• Balloon• 2 rulers

IntroductionHuman lung capacity can be measured in several ways. One way is by using a piece of laboratory equipment called a spirometer. However, lung capacity also can be measured by using a balloon. The data you obtain may not be as accurate as that obtained using a spirometer.

Several different lung volume measurements can be made. The largest possible amount of air which can be exhaled after drawing in a deep breath is the vital capacity. The amount of air that remains in the lungs after exhaling normally during normal breathing is about 500 cm3. This volume of air is called the tidal volume. A certain amount of air in the lungs cannot be expelled. This amount is the residual volume.

Procedure A: Vital Capacity1. Stretch a balloon several times.2. Take as deep a breath as possible. Then exhale all the air

you can into the balloon and pinch the balloon closed to prevent air from escaping.

3. Measure and record the diameter of the balloon in Column A of Table 1 (below). Use the figure to the right as a guide for measuring balloon diameter.

4. Deflate the balloon and run FOUR MORE. Record the diameter of the balloon for each.

Table 1: Balloon Diameters and Lung Volumes


Procedure Part B: Expiratory Reserve1. Exhale normally.2. Without inhaling as you normally would, put the balloon in your mouth and exhale all the air still left in your lungs.

NOTE: This step is different from what you did in Part A.3. Measure and record the diameter of the balloon in Column B.4. Run FOUR MORE trials. Record the diameter of the balloon for each trial.

Procedure Part C: Tidal Volume1. Take in a normal breath. Exhale into the balloon only as much air as you would normally exhale. DO NOT force

your breathing.2. Record the diameter of the balloon in centimetres in Column C.3. Run FOUR MORE trials.

Procedure Part D: Conversion of Diameters to VolumeLung volume is expressed in cubic centimetre units (cm3). (1000 cm3 is equal to a litre)

1. To convert from balloon diameter to volume, locate the balloon diameter on the horizontal axis of the graph on the next page. Follow this number up to the heavy line, then move across to locate the corresponding volume. For example, if your balloon diameter is 14.5 cm, then the corresponding lung volume is 1500 cm3. Use the dashed line on the graph as an example of how this procedure is done.

2. Convert each diameter for vital capacity, tidal volume, and expiratory reserve to volume.3. Record the volumes in Column D, E and F in the table below.4. Calculate and record for average lung volume for each of the 3 measurements.


Analysis1. Define the following terms:

a) vital capacity

b) expiratory reserve volume

c) tidal volume

2. Using your average volume measurements in the table, record your measured..

a) vital capacity

b) expiratory reserve volume

c) tidal volume

3. The following values were obtained through the use of a special machine called a SPIROMETER. Note

that these are average values.

a) How does your average vital capacity compare to the value obtained by a spirometer?

b) Why might these numbers not agree?

c) How could you improve the accuracy of this experiment without using a spirometer?

4. A close relationship between height and vital capacity exists. Complete this chart using your height for

Column A and one of the following factors for Column B:

20 for females 22 for female athletes 25 for males 29 for male athletes

a) Are your calculated and experimental values the same? Explain.

5. a) What is your breathing rate for 1 minute?

(Measure the number of times you breathe in or out in 1 minute)

b) How much air (in cm3) do you inhale in 1 minute?

(HINT: Use your average tidal volume from your table)

MALE FEMALE

Vital Capacity 5000 cm3 4000 cm3

Expiratory Reserve 1200 cm3 1000 cm3

Tidal Volume 525 cm3 475 cm3

AYOUR HEIGHT IN CM

BFACTOR

CCALCULATED VITAL

CAPACITY (A x B)

hematology “the study of blood” microscope...

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