Physioex Lab 2 Review Sheet Define the Terms

Main Body

Claret Lab

Learning Objectives

• Explain hematocrit, including the significance of values exterior of the normal range.
• Decide hematocrit from a claret sample image.
• Explain the ABO and Rh blood groups and their clinical significance.
• Conduct blood typing on a synthetic-blood sample.
• Identify and draw all formed elements in a human being claret smear.
• Land the relative proportions of formed elements in human claret.
• Demonstrate skillful microscope use.

Blood Composition and Hematocrit

Composition of blood

Overview of Blood [link opens in new window]

1. 55% = Plasma
   1. Proteins (for claret pressure level, clotting, and immune functions)
   2. Water (92% of plasma)
   3. Electrolytes
   4. Hormones
   5. Nutrients
   6. Blood gases
   7. Waste
2. 45% = Formed Elements
   1. Crimson Blood Cells (erythrocytes)
   2. Platelets (thrombocytes)
   3. White Blood Cells (leukocytes)

Hematocrit

• Definition: The volume—reported equally a percent—of packed elements (mainly cerise claret cells) in a blood sample.
• Clinical relevance: Provides information well-nigh the oxygen-carrying chapters of blood. Depression hematocrit means less red claret cells conveying O2.
• Healthy ranges:
  • Male: ____________%
  • Female:____________%

Effigy ii.1. Composition of Blood by Open Stax / CC BY 4.0.

Effigy 2.ii. Measuring hematocrit by Ashley Kajioka / CC BY four.0

What is the hematocrit for this sample? ___________

Claret Typing

Blood Typing

Blood type refers to the presence or absence of specific molecules, called antigens, on the red claret cell (RBC) RBC surface. Antigens are molecules, such as proteins, lipids, carbohydrates or nucleic acids, that your body tin can utilize to differentiate self and non-self. People with different claret types accept dissimilar antigens on their RBCs.

Antibodies are produced in response to some antigens (not-cocky), and are mostly used by the immune organization to recognize and facilitate removal of objects (viruses, leaner, tumorous cells, etc.) that do not belong in the body.

There are more than than 50 blood types in the homo population. The near clinically significant are the ABO and Rh(+/-) claret groups.

The ABO Blood Grouping

A and B antigens are glycoproteins on the RCC surface. ABO typing does not affect a person's Rh (+ or -) designation. Figure 2.3 Blood Blazon

Type A blood: A antigens on cell; anti-B antibodies in plasma

Type B blood: B antigens on cell; anti-A antibodies in plasma

Type AB blood: both A and B antigens on cell; neither anti-A nor anti-B antibodies in plasma

Type O blood: neither A nor B antigens on cell; both anti-A and anti-B antibodies in plasma

If a different type of blood is put into your bloodstream, the blood will agglutinate (dodder) and hemolysis (bursting) occurs within the foreign blood cells. Agglutination due to antibodies and antigens is a different process than blood clotting, which involves fibrin and other cascades associated with hemostasis.

Figure 2.3. Blood Type by Open Stax / CC Past iv.0

Rh Claret Grouping

Rh antigens are named after the rhesus macaque, a primate with many blood similarities to homo. There are many Rh antigens in humans, merely the D type of Rh antigen is the most clinically significant. Considering of this, in claret typing, sometimes D and Rh are used interchangeably. The Rh factor is grouped with ABO claret group to identify a claret blazon (example A+, B-, O-).

Type Rh+ (positive) blood: Rh antigens on cell

Type Rh- (negative) blood: no Rh antigens on cell

Unlike ABO blood type, no anti-Rh antibodies are present in Rh- individuals unless they take been exposed to Rh antigens. If Rh+ blood is introduced into an Rh- individual, anti-Rh antibodies volition be produced confronting the Rh(+) blood.

Importance of Rh during Pregnancy

This is a critical consideration in pregnancy for Rh- mothers if the fetus is Rh+. If any of the Rh+ claret enters the mother'south circulation, the female parent'southward immune organization will produce anti-Rh antibodies that will hemolyze her baby'southward blood (and any future Rh+ fetuses). This is called hemolytic disease of the newborn or erythroblastosis fetalis. Information technology is prevented with RhoGAM, a dosage of anti-Rh antibodies, given to the mother at 27 weeks and inside 72 hours of giving birth in guild to destroy any fetal blood cells in her claret so she volition not produce her own anti-Rh antibodies. RhoGam antibody dosage is small enough non to hurt fetus, but potent enough to go on mom'due south immune system from attacking fetus. See Figure 2.4.

Effigy 2.four. Erythroblastosis_Fetalis past Open up Stax / CC By 4.0

Determining Blood Type

To determine claret types, antiserum is used. The serum contains antibodies that may react with antigens on the RBC surface.

If using anti-A antiserum (contains anti-A antibodies) and the blood sample agglutinates (clumps), this indicates the presence of A antigens.

Which blood types have A antigens?  Fill in the type. Type _____  and Type _____

If using anti-B antiserum (contains anti-B antibodies) and the blood sample agglutinates (clumps), this indicates the presence of B antigens.

Which blood types have B antigens?  Make full in the type. Type _____  and Type _____

If using anti-Rh (anti-D) antiserum (contains anti-Rh (anti-D) antibodies) and the blood sample agglutinates (clumps), this indicates the presence of Rh antigens.

Which claret blazon has Rh antigens?  Fill up in the blazon. Type _____

Blood Typing Data

Complete the blood typing on your samples and enter your data in Table two.1.

For each claret sample:

1. Place a drib of blood in each of the three depressions of i testing tray. Each depression has a label of A, B, or Rh(D). One tray is used for each blood sample.
2. Place a drop of the antiserum that is associated with each depression. For instance anti-A antiserum (containing anti-A antibodies) goes into the depression marked A. In that depression, you volition be testing to run into if the anti-A antibodies agglutinate RBCs with A antigens. Do the same for anti-B and anti-Rh sera into each of their depressions in the tray.
3. Stir the combination of the blood and antiserum in each depression with the color coded toothpick. Do not mix toothpicks across depressions.
4. Examine the samples for agglutination and make full out your data table to make up one's mind the claret blazon for each sample.

Using claret sample	Antiserum contains antibody Anti-A and tests for antigen A	Antiserum contains antibody Anti-B and tests for antigen B	Antiserum contains antibiotic Anti-Rh (D) and tests for antigen Rh (D)	What is the blood type?
Blood sample 1	Agglutinated? Yes or No	Agglutinated? Yes or No	Agglutinated? Yep or No	Blazon:
Claret sample two	Agglutinated? Yep or No	Agglutinated? Aye or No	Agglutinated? Yeah or No	Type:
Blood sample 3	Agglutinated? Aye or No	Agglutinated? Yes or No	Agglutinated? Aye or No	Type:
Claret sample 4	Agglutinated? Yes or No	Agglutinated? Yeah or No	Agglutinated? Aye or No	Type:

Table ii.1 Blood Typing Data

Homo Blood Microscopy and General Microscope Utilize

In this function of the lab you will employ a microscope to examine erythrocytes, leukocytes, and platelets. These three constituents are referred to as the formed elements of blood. Platelets are not considered a prison cell, as they are enclosed cytoplasmic fragments. A complete blood count with differential is a clinical mensurate that states the percentages of each claret cell type and is used for various diagnostics such equally determining anemia or types of infections or allergic reactions. Erythrocytes are the most numerous blood cell, and then the count of the unlike leukocytes goes from most to to the lowest degree numerous in this order: neutrophils, lymphocytes, monocytes, eosinophils, and basophils.

Formed Elements of Claret

Erythrocytes (Ruby Claret Cells)

Erythrocytes

See Figure 2.5 below and notice the numerous, round, pink cells in the background each of the leukocyte images. These are red blood cells (RBCs). Some look like they have a hole in the middle, merely this is due to the thin surface area of the biconcave shape that allows for flexibility and to increment expanse.

Primary function: transport respiratory gases to and from tissues.

Lack a nucleus.

Virtually arable of all blood cells.

Contains millions of Hemoglobin molecules: allow for binding of O2 and CO2.

Platelets

Platelets

Also called thrombocytes but not technically a cell. They are produced past the fragmentation megakaryocytes that are in bone marrow tissue.

Involved in coagulation: the process of clot formation.

During coagulation, molecules (fibrin) join to form long threads that course a cyberspace to trap platelets and plug the wound.

Leukocytes (White Blood Cells)

Leukocytes

See Figure 2.v below.

Only formed elements with a nucleus.

Lacks hemoglobin.

Travel between endothelial cells of capillaries and tissues.

2 types of leukocytes: granular and agranular.

Effigy 2.5. Leucocyte Key by Open Stax / CC BY 4.0

Granulocytes

See Effigy 2.6 below.

All take granules in cytoplasm.

1. Neutrophils (40-threescore% of total white blood cell (WBC) count)
   1. most common WBC
   2. 1st to get in at wound/infection site
   3. release cytotoxins
   4. capable of phagocytosis
2. Eosinophils (1-4% of total WBC count)
   1. phagocytize microbes that immune organisation has coated with antibodies
   2. decrease inflammatory response at site of wound
3. Basophil (0.5-1% of WBC count)
   1. release histamines (crusade vasodilation) and heparin (prevents clotting)
   2. important in allergies

Figure 2.6. Granular Leucocytes by Open Stax / CC BY 4.0

Agranulocytes

Refer dorsum to Effigy two.5

Fewer and less obvious granules in cytoplasm.

1. Monocytes (ii-eight% of total WBC count)
   1. wanderers, patrol body tissue for microbes and worn-out tissue cells
   2. 2nd to arrive at wound site
   3. phagocytize dead cells/debris that has accumulated at site of wound/infection
2. Lymphocytes (20-40% of WBCs)
   1. smallest leukocyte, arable in bloodstream, occur in lymph nodes and glands
   2. specialized lymphocytes:
      1. T-cells: attach to and destroy infected or cancerous cells past releasing cytotoxic molecules and secreting antiviral/proinflammatory molecules
      2. B-cells: manufacture antibodies that attach to foreign pathogens/cells and help destroy them
      3. Natural Killer cells: can detect sick, malignant, and infected cells and release cytotoxic molecules to destroy them

Follow the instructions below for microscope use, and examine a human being blood smear. Sketch each of the formed elements of blood as seen in your view.

Microscope Parts and How To Handle Them

In that location are many different types of microscopes. We shall learn virtually the chemical compound low-cal microscope. It uses visible light to visualize the specimen, and passes that light through two split up lenses to magnify the image. Compound microscopes accept a lot of moving parts and can be damaged and broken through mishandling. A large part of learning how to utilise the microscope properly involves learning how to avoid damaging it. To do that, you beginning have to be familiar with the parts.

In Figure 2.7, in that location are 2 compound microscopes shown with key parts identified. The one on the left is monocular and the one on the right is binocular. Many of the parts of the two microscopes are in slightly unlike locations.

When you first sit in forepart of a microscope, take a moment to discover the key parts, especially the knobs for focus, condenser aligning, and stage control. When viewing a specimen, your optics will be at the eyepieces (oculars), and if y'all take hold of the wrong knob by accident, you can lose your paradigm or damage the microscope.

Effigy 2.7 Fundamental parts of a compound light microscope. Compound low-cal microscope by Ross Whitwam / CC BY SA

Eyepiece (Ocular)

The eyepiece contains the eyepiece lens, one of the 2 lenses doing the magnifying in a compound microscope. If the microscope is binocular, use both eyepieces, adjusting them to ensure they fit the spacing of your eyes. For successful binocular viewing, bring your prototype into focus with the lowest power objective, while looking through only the non-adjustable ocular. Then while looking only through the adaptable ocular, rotate its focus band to bring that ocular into clear focus. Now the paradigm should be articulate as you expect through both oculars.

Carrying arm

When moving a microscope, even if it is simply a few inches, ever selection it up by the carrying arm. Do not drag the microscope: selection it up. The microscope will have prophylactic feet that prevent it from sliding, and then if you try to drag it, it will shake and vibrate and possibly damage parts. Never pick up the microscope by any part other than the carrying arm. The other parts are by and large much more fragile and prone to breaking.

Objective lenses

Well-nigh compound low-cal microscopes will contain three to four objective lenses that can exist rotated over the slide. Sometimes these lenses are only chosen objectives. When a detail objective has been fully rotated into position, you will feel a click as that objective locks into identify. The objective lens is the second of the two lenses doing the magnifying in a compound microscope, so if it is not snapped into proper position, you lot will not meet the proper image. Each objective lens can commonly be unscrewed from its position in the rotating turret that houses it, so be conscientious y'all are rotating the turret, not unscrewing an objective. Do non unscrew the objectives from the turret. Each objective lens has a different magnifying power, then the prototype on your slide volition exist magnified to bottom or greater extents, depending on which objective lens you have chosen. Each objective's magnification power will be written somewhere on the side of the objective.

Phase and stage clips

The slide will be held in identify on the stage with stage clips that printing against the sides of the slide. The clips do not sit down in a higher place or beneath the slide. They are jump-loaded to concur the slide edges and lock the slide in identify so that the stage controls can move the position of the slide smoothly.

Stage controls

These allow you to motility your slide while you are viewing it, but only if the slide is properly clipped in with the stage clips. Find the stage command dials on your microscope before you lot start viewing your slide. There are two dials—one moves the slide left and right, the other moves the slide upwardly and down. Notice in Figure 2.7, the dials are on top of each other and below the stage on the binocular microscope, however, they are 2 separate dials and above the stage on the monocular microscope.

Fibroid focus

This is the larger of the two focus knobs. Use it with the lowest ability objective to get the specimen approximately in focus. Afterwards that, only utilize the fine focus knob, even after you lot change to a higher-ability objective. Observe in Figure ii.7, the binocular microscope fine focus knob is surrounded by the coarse focus knob, yet, the monocular microscope coarse focus and fine focus knobs are separated.

Fine focus

This is the smaller of the two focus knobs. This is the focus you will use repeatedly in viewing slides in one case they are focused with the coarse focus.

Condenser position adjustment

You typically will non demand to adjust this knob. It controls how far the light condenser is from the slide, which should be properly adjusted before you employ the microscope.

Condenser opening adjustment (not shown in effigy)

This opening can be adjusted, usually by rotating a ring around the condenser. Be sure this has not been left airtight by a previous user. Experiment with dissimilar opening sizes to make up one's mind what is all-time for your specimen.

Iris diaphragm lever

Find the lever under the stage where calorie-free passes through to the slide. It opens and closes an iris to let more or less calorie-free through the slide. In some specimens there is not much contrast betwixt the colours and shades of the different components being magnified. Changing the view past adjusting the iris tin can allow y'all to better run into some of the details you are trying to magnify.

Rheostat: Light intensity (not shown in figure)

Rotate this dial to adjust the effulgence of the calorie-free source. Plow this to its a depression setting earlier looking through the eyepieces. Yous may need to increase the intensity every bit you increase the ability of your objective. Turn the rheostat all the mode downwardly before turning off your microscope.

Lab exercises

Carry out the activities listed beneath and answer the questions.

1. Option up your microscope and bring it shut enough that you can look into it comfortably from where you are sitting with healthy posture. Arrange it so that the stage is facing you and the eyepieces are rotated toward you. What office of the microscope did you take hold of in order to pick it up and move it?
2. Where are the locations of the two stage adjustment knobs on your microscope?
3. Where is the location of the coarse focus knob?
4. Where is the location of the fine focus knob?
5. Is there a condenser opening adjustment ring?
6. Detect the diaphragm lever. Looking in the hole in the heart of the phase, what happens when you move the diaphragm lever in each direction?
7. Afterwards cleaning a slide equally instructed by your professor, place the slide on the stage.
8. Have the steps described in the ocular department to obtain clear view through both of your oculars. If you article of clothing glasses, attempt with and without to determine which is best for you.
9. Adjust the condenser opening and iris lever. How does this modify your view?

Checking out and storing the microscope

When you finish your microscope work with the blood slide, exist prepared to have your instructor check off each of these items before putting away your microscope.

1. Turn the rheostat to its dimmest setting.
2. Turn off the ability, unplug, and wrap the string around the base.
3. Wipe the objective lenses with methanol and lens paper. Find this is lens paper, non kimwipes. Using any newspaper other than lens paper tin scratch the lens.
4. Rotate the objective lens turret so the everyman power objective faces downward.
5. Wipe the stage clean with a kimwipe and move it to the lowest position.

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"Claret Lab" is MODIFIED from:

• Beefcake and Physiology by Open Stax / CC BY 4.0. Download for free at http://cnx.org/contents/14fb4ad7-39a1-4eee-ab6e-3ef2482e3e22@12.viii.
• An Overview of Blood, Beefcake and Physiology by Open up Stax / CC Past four.0. https://cnx.org/contents/FPtK1zmh@12.viii:IUrEdFyf@10/An-Overview-of-Claret
• UGA Anatomy and Physiology 2 Lab Manual by University System of Georgia / CC Past 4.0
• Human Anatomy and Physiology Lab (BSB 141) past Lumen Learning / CC BY-SA
• A&P Labs. Authored by: Ross Whitwam. Provided by: Mississippi University for Women. Located at: http://www.muw.edu/. License: CC Past-SA: Attribution-ShareAlike
• Labeled compound light microscope. Authored by: Ross Whitwam. Provided by: Mississippi University for Women. Located at: http://www.muw.edu/. License: CC By-SA: Attribution-ShareAlike

Citation notes:

• Hesse, DeLoris; Cozart, Deanna; Szymik, Brett; and Nichols, Rob, "UGA Anatomy and Physiology 2 Lab Transmission" (2017). Biological Sciences Open Textbooks. 14.
  https://oer.galileo.usg.edu/biology-textbooks/14

"Microscope Parts and How to Handle Them" is MODIFIED from:

• Lumen Learning. CC By-SA https://courses.lumenlearning.com/ap1x9x1/affiliate/the-parts-of-a-chemical compound-microscope-and-how-to-handle-them-correctly/
• A&P Labs. Authored by: Ross Whitwam. Provided by: Mississippi University for Women. Located at: http://world wide web.muw.edu/. License: CC Past-SA: Attribution-ShareAlike
• Labelled compound light microscope. Authored by: Ross Whitwam. Provided by: Mississippi University for Women. Located at: http://www.muw.edu/. License: CC BY-SA: Attribution-ShareAlike

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Source: http://pressbooks-dev.oer.hawaii.edu/shook/chapter/chapter-2-blood-lab/

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