Anatomy & Physiology 2002 Lecture Notes, Directory

These are lecture notes listed in sequence:

001a_Telencephalon.Nov15

001b_Autonomic_NS.Jan11

001c_Autonomic_neurotransmitters.Feb10

001d_Emotions_Memory.Feb11

01_Olfaction_Taste.Jan16

001d_Integration_Emotions.Jan16

02_Eye_Accessory_Structures.Jan16

03_Eye_histology_physiology.Jan16

04_Hearing.Feb12

05.5_Endocrine_Intro.Feb16

05_Balance_Organs.Feb16

06_Pituitary.Feb16

07_Thyroid_Adrenal.Feb2016

08_Pancreas.Mar11

09_Pineal.Feb2016

10_Blood.Feb2016

11_12_Leukocytes.Hemostasis.Feb2016

12_Blood_Clotting.Feb2016

13_Heart.Mar2016

14_Heart_electrical.Mar2016

15_Arteries_and_Veins_Lecture.Mar2016

16_Circulation_Special.Mar2016

17_Lymphatic_Sys.Mar2016

18_Defenses.Mar2016

19_Immunity.Mar2016

20_Respiratory_Sys.Mar2016

21_Respiratory_Physiology.Mar2016

22_Digestive_Sys_to_stomach.29Mar2016

23_Digestive_Sys_Intestines.Mar2016

24_Liver_&_Pancreas.March2016

25_Metabolism_I_catabolism.April2016

26_Metabolism_II_BMR_hunger.11April2016

26a_Nutrition_I_water_soluble.Apr10

26b_Nutrition_II_Oil_Soluble_Sources.Apr10

27_Urinary_System.12April2016

28_Kidney_regulation.12April2016

29_Repro_embryology_spermatogenesis.12April2016

30_Male_reproductive_system.12April2016

31_Female_Repro_Anat.12April2016

32_Female_Hormones.12April2016

33_Development.June09

34_pregnancy_birth.June10

35_Labor_May12

Blood Pressure Determination

Blood Pressure Determination

Blood Pressure Determination(See Merck Manual, 17th Ed, p 1603-1604, 1629-1635.)

The cyclic beating of the heart pumps blood to the lungs via the pulmonary circuit, and then to the body via the systemic circuit. The pressure in the arteries alternates from a maximum when the heart is in full contraction, called systole (Greek for contraction), to a minimum when the heart is relaxed, called diastole. Blood pressure (BP) is determined by measuring these pressures by first constricting blood flow using a sphygmomanometer (an inflatable cuff equipped with a manometer to display the pressure), and then monitoring the flow of blood through the artery with a stethoscope as the cuff pressure is allowed to slowly decline. The pressure at which the first thumping sounds of blood flowing under the cuff marks systole, and the point at which these sounds become muffled is diastole. (The sphygmomanometer is no longer able to close off the artery .)

The average BP in young adults is 120/80 mm Hg, while the upper limit in healthy individuals is 140/90. Individuals with BP above this but below 180/115 are said to have moderate hypertension. BP >180/115 constitutes hypertension. Hypertension, besides forcing your heart to work harder, can impact your circulatory system in much the same way as over-pressurization of a tire: it increases the likelihood of a blowout. Such a rupture is termed a hemorrhage and can cause blindness, kidney failure, or paralysis according to what tissue has its circulation interrupted.

Arteries normally act as blood pressure shock absorbers by stretching during systole. If the arteries toughen or harden, a condition called arteriosclerosis, the loss of elasticity causes the transmission of unmoderated pressure through the arterial system. Hypertension routinely occurs as a function of aging, but diet also plays an important role. Reduction of animal fat is important in ameliorating the sclerotic process, as is regular physical exercise. Reduction of salt intake can also lower BP.

This lab requires QUIET during the activity.

EQUIPMENT:
Stethoscope (Wipe earpieces with 70% EtOH to avoid transmission of ear infections.)
Sphygmomanometer (Illustrate and label all bolded functional parts. )

PROTOCOL:

1) Have subject seated comfortably, with non-writing arm bare, unconstricted and supported on a table.

earpieces_P4013656
Stethoscope ear pieces point forward

2) Insert ear pieces The ear pieces should point forward to conform with your ear canals.

adjust_nut_P4013658
Practice fine control of air control nut

 

3) Adjust valve nut for snug closure, but not too tight

tuck_stetho_P4013659

4) Fit sphygmomanometer cuff loosely above elbow so the manometer is easily visible. Insert the diaphragm of the stethoscope under the cuff, over the brachial artery (front of arm).

 


5) Pump up cuff to around 100 mm Hg first and confirm that you can hear heart sounds. Then pump further so that no heart sounds are heard, approximately 150-160 mm Hg for a resting young adult. (Do not go to painfully high pressures of >200 mm Hg unless necessary.)

adjust_nut_P4013658
Practice fine control of air control nut

6) Slightly loosen valve nut so that pressure drops slowly (the manometer’s needle moves lower).


7) Listen carefully for the first heart sound to appear. Note the pressure at which this occurs. This is the systolic pressure. (The needle will begin to pulse just before the sounds can be heard.)

8) Allow the pressure to continue to slowly drop and note the pressure at which the beats become muffled or indistinct, called the diastolic pressure.

tuck_stetho_P4013659
9) Allow cuff to deflate completely to reestablish circulation, record the BP just determined, then repeat a second and third time. Average the three systolic and the three diastolic pressures.
Repeat these measurements after physical exertion, administration of various agents (caffeine, sugar, alcohol, etc) to determine their effect on blood pressure.

White Blood Cell Count

White Blood Cell Count

See related protocols: Blood Cell Counts, Blood Typing, Hematocrit.

White blood cells (WBC), or leukocytes, are involved in fighting infections and clearing away dead cells.  During infections, their numbers increase dramatically above the normal range of 4,800 to 10,800 WBC/cu mm.  Finding WBC content above 10,800 is suggestive of infection, or other problems (such as leukemia).

 Leukocytes are without pigment, and must be stained to be counted.  Therefore, the diluent contains crystal violet for this purpose.  It also contains acetic acid which fixes the WBC, and causes the lysis of RBC so they are not visible.  Note that a different dilution pipette is used than that in the RBC technique.

Each desk should assemble the following:

blood_letting_setup_P3130052

Image of desk set up for blood studies Equipment and supplies

Blood letting set up
2 hemacytometers
2 coverslips
one autolet
2 platforms (for drawing blood)
2 lancets (for drawing blood)
1 250 mL beaker for waste fluid
1 bottle WBC diluent (purple)
1 bottle RBC diluent (clear)
2 Kimwipes, soaked in 70% EtOH
2 paper towels

Blood cell counts can be performed using the hemacytometer. This is a precision instrument possesses a platform with microscopic grid scoring above which a specified quantity of fluid is held. By properly diluting blood, counting all cells in specified squares, and multiplying by the proper conversion factor, the number of cells per cubic millimeter can be determined.

Because of the potential dangers of working with blood, we will first practice the necessary dilutions and use the hemacytometer to count yeast cells. Be certain to master these skills before you attempt to do the blood work.

First illustrate:
1) the dilution pipets, explain their use and what the dilution factors would be
2) the grids for WBC counts
3) the grids for RBC counts.  Practice drawing water up in the pipet to the desired volume several times.
Practice drawing water up to the 0.5 and 1.0 volumes several times so that you are confident of your skill.  You will then dilute the suspensions of yeast, place an aliquot on the hemacytometer, and count all yeast in five designated squares. The convention is to count all cells touching left and bottom sides, ignore cells touching top and right sides.  When the five squares are counted, you add them up and multiply by the appropriate dilution factor (see below).

When finished for the day, wash out the pipettes and hemacytometer thoroughly with soap and water, rinse well, finish with distilled H2O rinse, replace in case.

PROTOCOL (These steps are similar to RBC, read that protocol carefully

OLYMPUS DIGITAL CAMERA

1. Swab the tip of a little-used finger with 70% EtOH.


2. Lance with quick, firm jab to the side of the pad of the finger, wipe away first blood.

OLYMPUS DIGITAL CAMERA
3. Using dilution pipet with the WHITE mixer, draw up to the 0.5 mark. Do not allow blood to congeal in pipette! Proceed immediately to the next step:

OLYMPUS DIGITAL CAMERA
4. Fill the pipet to the 11 mark with crystal violet diluent. (See below for formula.)

07_filled_with_diluent_P3042339
5. Shake well to mix with the tip sealed with your finger.


6. Empty ~1/2 of pipet into waste container, add a small amount of the diluted blood apply to the second chamber of the hemacytometer. It should flow in to fill the chamber. (Do not over fill).
If the chamber is overfilled, quickly remove the overflow with a paper towel.


7. Let the preparation sit for a minute (for cells to settle).

13_WBC_stained_100x_P3113670
8. Examine under 100x, count the five fields indicated squares of blue-stained WBCs with a clicker (fields: top L & R, bottom L & R, center). Include in the count all cells touching left and bottom sides, ignore cells touching top and right sides.
Here are other pictures of
WBCs
WBCs
9. CLEAN UP THE EQUIPMENT: Wash out the hemocytometer, pipettes and mouth pieces thoroughly with soap and water, rinse well, finish with distilled H2O rinse, replace in case. Replace along with two pieces of hose in the case, return to the proper location in the drawer.
10. Calculate the WBCs/cmm: sum the 5 groups, multiply by 40. Enter your blood cell counts in the class data table

These are RBC views which will have to suffice until we have some WBC views for illustration…


1 Diluent for white blood cells:

10 mg crystal violet
1.0 ml glacial acetic acid
q.s. to 100 mL with d H20

Red Blood Cell Count Using a Hemacytometer

Red Blood Cell Count Using a Hemacytometer

See related protocols: Blood Cell Counts, Blood Typing, Hematocrit.

SAFETY NOTE:
Avoid infection with blood-borne pathogens such as HIV and hepatitis B, by using protective gloves when handling other peoples’ blood.  Dispose of the blood-contaminated sharps in the marked container, and the rest as indicated by the instructor.

INTRODUCTION:
Sufficient numbers of red blood cells (RBC, or erythrocytes) are necessary for adequate transport of oxygen from the lungs to the peripheral tissues.  Too few RBCs constitutes a pathological condition known as anemia (lit., without blood).  According to the Merck Manual, normal values of RBC/cmm for males is 5.4 + 0.8 million, and 4.8 + 0.6 for females.  Anemic levels for adult males are below 4.5 million, for females below 4.0 million.  We will perform red and white blood cell counts on your blood in the lab using a hemocytometer and appropriately diluted blood.

Blood cell counts can be performed using the hemacytometer. This precision instrument possesses a platform with microscopic grid scoring.  Rails on either side hold up a cover slip so that a specified quantity of fluid is held. By properly diluting blood, counting all cells in specified squares, and multiplying by the proper conversion factor, the number of cells per cubic millimeter can be determined.

PRELIMINARIES:
You should have already illustrated the following when you performed the practice counts using yeast last week:
1) the dilution pipets, with explanations of their use and what the dilution factors would be
2) the grids for WBC counts
3) the grids for RBC counts. Review the illustration of the five hemacytometer fields which you drew your notebook.

The red blood cell count may be performed at the same time as the white blood cell count and hematocrit, if you get blood to blow adequately.

EQUIPMENT SUPPLIES
2 hemacytometer kits, each with:
1 hemacytometer
1 coverslip
1 WBC diluter pipet with hose and mouthpiece
1 RBC pipet with hose and mouthpiece
1 autolet
2 lancet needless (for drawing blood)
2 platforms (for drawing blood)
microscope
squirt bottle with 70% ethanol
Kimwipes (to be soaked in EtOH)
1 bottle WBC diluent (purple)
1 bottle Ringer’s Solution (a clear diluent for RBC)
1 250 mL beaker for waste fluid
paper towel

RED BLOOD CELL COUNT:
Set up all equipment on your desk so that you are sure to have everything at your finger tips for the procedure.
Prepare an autolet with a sterile lance and platform.
1. Swab towards the side of the tip of a little-used finger with 70% EtOH. (NOT close to the cuticle!)
2. Lance by placing the platform of the autolet against the finger tip and pressing the trigger. Alternatively, us a lancet with quick, firm jab to the side of the pad of the finger. Wipe away first blood.
3. Using the dilution pipet with RED mixer from hemacytometer kit, draw blood up to the 0.5 mark. This is best done by slightly slanting the pipette down to allow blood to flow in. (Do not allow air to be drawn into the pipet or it will not draw the correct volume of blood. Slight suction should start it. (Make sure the hose is not kinked shut.) Keep the pipette level once you have filled it. Do not allow blood to congeal in pipette! Immediately proceed to the next step:
4. Continuing to hold the pipet as horizontal as possible, draw Ringer’s solution diluent up to the 101 mark. (Dilution of 1 to 200.)
5. Seal the tip with your finger and shake well to mix.
6. Empty ~1/2 of pipet into waste container
add a small amount of the diluted blood to one chamber of the hemacytometer to just fill the chamber of the hemacytometer. It should flow in to fill. (Do not over fill).
7. Let the preparation sit for a minute (for cells to settle).
8. Center the grid at 100x, switch to 400x and count and record the RBCs in each of five fields (each with 16 smallest squares) with a clicker (fields: top R & L, bottom R & L, center). Include in the count all cells touching left and bottom sides, ignore cells touching top and right sides.
Calculate the RBCs/cmm by adding the cells in the 5 groups and multiplying by 10,000 (i.e., add four zeros). Enter your RBCs/mm in the class data table.
9. Wash out the pipette thoroughly with soap and water, rinse well, finish with distilled H2O rinse, replace in case


Ringer’s Solution, per 100 mL:

860 mg NaCl
30mg KCl
35mg CaCl2

dissolve in dH2O and q.s. to 100 mL

Blood Cell Count Practice

Blood Cell Count Practice/Hemacytometer And Diluting Pipette Practice

See related protocols, Blood Cell Counts, Blood Typing, Hematocrit.

Blood Cell Count Practice

Hemacytometer with cover slip
WBC diluting pipet (white), hose and mouthpiece
RBC diluting pipet (red), hose and mouthpiece
yeast suspension:
For WBC simulation:  50 mg dry baker’s yeast + 100 mL water
For RBC simulation:   1 package baker’s yeast + 100 mL water
WBC diluent
RBC diluent
50 mL beaker for waste fluid
paper towel

Blood cell counts can be performed using the hemacytometer. This is a precision instrument possesses a platform with microscopic grid scoring above which a specified quantity of fluid is held. By properly diluting blood, counting all cells in specified squares, and multiplying by the proper conversion factor, the number of cells per cubic millimeter can be determined.

Because of the potential dangers of working with blood, we will first practice the necessary dilutions and use the hemacytometer to count yeast cells. Be certain to master these skills before you attempt to do the blood work.

First illustrate:
1) the dilution pipets, explain their use and what the dilution factors would be
2) the grids for WBC counts
3) the grids for RBC counts.  Practice drawing water up in the pipet to the desired volume several times.
Practice drawing water up to the 0.5 and 1.0 volumes several times so that you are confident of your skill.  You will then dilute the suspensions of yeast, place an aliquot on the hemacytometer, and count all yeast in five designated squares. The convention is to count all cells touching left and bottom sides, ignore cells touching top and right sides.  When the five squares are counted, you add them up and multiply by the appropriate dilution factor (see below).

When finished for the day, wash out the pipettes and hemacytometer thoroughly with soap and water, rinse well, finish with distilled H2O rinse, replace in case.

PRACTICE EXERCISE FOR LEUKOCYTE COUNT:

1. Suspend 50 mg dry yeast in 100 mL water.
2. Perform the dilution twice, add sample of each dilution to hemacytometer chambers 1 and 2 as spelled out in steps 3-6:
3. Using dilution pipet with the WHITE mixer, draw yeast suspension up to the 0.5 mark. Dab with piece of paper towel if needed to adjust volume. Proceed immediately to the next step:
4. Fill the pipet the rest of the way to the 11 mark with crystal violet diluent1. (This constitutes a 1:20 dilution)
5. Shake well to mix with the hose end sealed with your finger.
6. Empty ~1/2 of pipet into waste container, add a small amount of the diluted yeast to the first chamber of the hemacytometer. It should flow in to fill the chamber. (Do not over fill).
7.  Repeat steps 3-6 and fill the second chamber.
8. Let the preparation sit for a minute (for cells to settle).
9. Examine under 100x, count the five fields indicated squares of blue-stained yeast with a clicker (fields: top L & R, bottom L& R, center).
10. Calculate the WBCs/cmm: sum the 5 groups, multiply by 40. (Should be about 8,600 yeast cells/cmm)

RED BLOOD CELL COUNT PROCEDURE:
1. Mix a package of baking yeast with 100 mL of water, stir for 10 minutes to suspend.
2. Using the dilution pipet with RED mixer from hemacytometer kit, draw suspension yeast up to the 0.5 mark. This is best done by slightly slanting the pipette to allow the suspension to flow in. Slight suction should start it. (Make sure the hose is not kinked shut.) Keep the pipette level once you have filled it. Immediately proceed to the next step:
3. Draw Ringer’s solution diluent up to the 101 mark. (Dilution of 1 to 200.)
4. Shake well to mix with the hose end sealed with your finger.
5.  Empty ~ 1/2 of the pipet into the waste container, then add a small amount of the diluted blood to one chamber of the hemacytometer. It should flow in to fill. (Do not over fill).
6. Let the preparation sit for a minute (for cells to settle).
7. Center the grid at 100x, switch to 400x and count five fields of 16 smallest squares RBCs with a clicker (count these fields: top R& L, bottom R & L, center).
8.  Calculate the number of yeast/cubic millimeter: sum the 5 groups, multiply by 10,000 (i.e., add four zeros).
9. How many in the entire package?
10. CLEAN UP THE EQUIPMENT: Wash out the hemacytometer, pipettes and mouth pieces thoroughly with soap and water, rinse well, finish with distilled H2O rinse, replace in case. Replace along with two pieces of hose in the case, return to the proper location in the drawer.

Count as for WBC count. (this suspension is less viscous than blood)

Grids for WBC and RBC counts:

1 Diluent for white blood cells:

10 mg crystal violet
1.0 ml glacial acetic acid
q.s. to 100 mL with d H20

blood_letting_setup_P3130052

BLOOD LETTING SET UP PER DESK
13-Mar-08


2 hemocytometers, clean and polished
2 clean coverslips
2 WBC (white) diluting pipets with mouth piece and tubing
2 RBC (red) diluting pipets with mouth piece and tubing
1 bottle RBC diluent (clear)
1 bottle WBC diluent (purple)
2 hematocrit tubes, heparinized
1 crit-o-seal
2 self contained lancets
2 cotton balls
1 70% EtOH in squirt bottle
1 folded paper towel, torn in half, half per person
1 50 mL beaker for waste
1 250 mL beaker with about 50 mL dilute warm soapy water
2 clickers