A&P 2 Blood Lab Guide

Pre-Lab Information and Exercises Have someone in your group read the following out loud, while the others read along:

In this Pre-lab, we are going over some basic concepts regarding blood & immunity. These notes cover important concepts found in the lab videos. NOTE: the current videos are “old videos”, and so the notes may not align exactly with the videos. If you have any questions as to terms, etc., ask the teacher or your SI. NOTE: You will be tested on these concepts in BOTH LECTURE AND LAB. Some of the material will be covered in both! Some semesters, “Blood Types and Immunity” is covered in lecture, other semesters in lab. The notes seen in this guide also appear in Shuster’s Lecture Notes, so do not be surprised by the repetition! At the end of this guide, there are some review questions that you may do to assess yourself. Some of the pages have assessments using images and questions from McGraw-Hill Publishers. NO…I am not going to simply give you the answers. However, you can ask questions before the next exam/quiz. If there are some questions you can’t answer before lab, highlight them and see if you can answer them after lab!

Step 1. NOTE: T - First a re

- Blood ceWBCs use

* m

* ou * T Afoce

- In generown cells,

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- There ar

* pa * sy

Tth

UnderstanThese are the

eview of somePathoSymptInfectiAutoim

ells, like all cee these to ide

ANTIGEN - pmembrane.

* Theygenetihave. * Dete

ANTIBODIESut and attack

AGGLUTINOhese antibod

Agglutination:oreign cells froells are then p

Pus is a pimpaccummacro

ral, you do no, or you would

Autoimmune wn tissues.

* We w

d if agglutinaton.

re many antig

This way, weast immunity.

The only persystem, is you

Tissue rejectihreat. We wo

nding BLOOe same (or si

e important telogy or diseatoms (includion (bacteriammune disea

ells, have MARentify “self” fro

protein marke

y act as little “cally determin

rmine ”self” fr

S - immune syantigens.

OGEN - someies are “sticky

this “clumpinom causing dphagocytized

a protein-richple or pustule

mulation of thephages, form

ot make antibod destroy you

disease: a p

will see sever

tion happens

gens on the ce

e are protecte

son who sharr identical twi

ion: This sysould like to be

OD TYPES &milar) to the

erms we learnase

ding fever, dial, viral, fungase

RKER PROTom “non-self”.

ers; use to ide

flags” for idenned which typ

rom “non-self

ystem protein

e antigens proy”, and stick t

ng together”, damage in the, and destroy

h fluid that lea, formed at th

e old, destroyeming the visco

odies against ur own tissues

person’s immu

al in the follow

in the blood s

ell’s surface.

d against a fo

res all your ann, if you have

stem was desable to put fo

& IMMUNITYnotes found

ned in A&P1 (

arrhea, and al, protozoan

EINS on the e.

entify “foreign”

ntification, pe you will

f”.

ns that seek

omote agglutinto each other.

which prohibe body. Aggluyed.

aves the bodyhe site of inflaed foreign anus pus.

the antigens s!

une system a

wing chapters

stream after a

oreign cell ma

ntigens, and we one. They a

igned in natuoreign tissues

d in Shuster’

(make sure yo

inflammation, parasitic,

external side

” cells. Speci

nation. .

bits the utinated

y through ammation duri

d immune ce

on your

attacks its

s.

a

atching our an

whose cells ware exactly th

re, where anys in the body,

s “Lecture N

ou know thes

n) etc.)

of their plasm

ial glycoprote

ing infection. ells that have

ntigens, and t

would not set he same gene

y foreign cell however. So

Notes” series

se terms):

ma membrane

eins on plasm

It is an been destroy

thereby gettin

off your immuetically!

is consideredo we must

s.

e;

a

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ng

une

d a

unR Nrego B Tanfo

*

A) THE A - This is aprotein m The immuprotein a there will

nderstand theRejection is wh

Not all antigenesponse. ABOoing to give s

Blood typing:

Titer: measurn immune res

or a time, to p

We will look

ABO GROUP

an antigen (glyight exhibit.

une cells checperson has abe agglutinat

e system to ghen the immu

ns are equalO is very reac

someone a blo

: analysis of t

e of the amousponse (possrevent a re-in

k at a couple

ycoprotein on

ck to make sut this site is gion.

et around it inune system at

lly reactive. Sctive, howeveood transfusio

the antigens p

unt of circulatible infection)

nfection. How

of important

n the plasma m

ure the cell haenetically det

n the case of ttacks an orga

Some incorreer, so we muson.

present.

ting antibodie). A person’s w long they ar

t blood group

membrane).

as the correct termined, so

a transplant, an or tissue th

ct antigens dost make sure t

s in plasma. blood will retre retained de

ups:

“ABO” refers

t version of “Ait is a test to s

such as a blohat we have t

o not always this one is co

Indicates thetain the circulepends....wee

s to the variati

ABO”. The vesee if this cel

ood transfusiotransplanted.

cause an immompatible if we

e person has ating antibod

eks, months, y

ons that this

ersion of the l “belongs”. I

on.

mune e are

had dies years.

f not,

* th

* th

* “Tis * he“U

- Ato

A person’s Rhe ability to m

A person’s Rhe ability to m

A person’s RTYPE AB”) ans called the “U

A person’s Re is “TYPE OUNIVERSAL D

Note th It woulin your

Agglutinationo a blood sam

RBCS might hamake the “B” a

RBCS might hamake the “A” a

RBCS might hand has no antUNIVERSAL R

RBCS might ha”) and has theDONOR” sinc

It doesn’t mis not antige

hat the “O” is

ld have been r head!

is easy to semple:

ave the “A” prantibodies - at

ave the “B” prantibodies - at

ave both the tibodies for thRECIPIENT”.

ave neither te BOTH antibce his blood h

matter which aen for them to

actually “Zer

more intuitive

ee, so it is eas

rotein at this sttacks blood t

rotein at this sttacks blood t

“A” and “B” pe ABO group

he “A” NOR tbodies - attachas no antigeantibodies areo stick to!

ro”, although w

e if they had c

sy to determin

site (in whichthat has the “B

site (in whichthat has the “A

proteins at thisp - cannot atta

the “B” proteicks ALL bloodns to attack.

e floating arou

we pronounce

called it AB-z

ne blood type

case, he is “B” protein “fla

case, he is “A” protein “fla

s site (in whicack a non-ma

ins at this sited put into him, und for this si

e it “oh”.

zero”, and I su

e visually by a

TYPE A”) andag”.

TYPE B”) andag.

ch case, he isatching ABO, a

e (in which ca, but is the

te, because t

uggest you do

adding antibod

d has

d has

s and

ase,

there

o that

dies

B) Rh BLO - Another the Rh promonkey, aexperimenbut the mo 1. The pecannot ma 2. The pe“anti- Rh” Rh factor. PLEASE “common“D”, along *Most comfetus - ER (i) We neimmune s

-

- -

(ii) Erythrfetus. Se

- fe

(iii) Only hRh+ child

-

- - - ba - fe

OOD GROUP

site on the plotein site (Rhas they sharents were perfoost common

erson HAS theake “anti- Rh”

erson lacks it if they come

.

NOTE: this ily-referred to”

g with its corre

mmon case ofRYTHROBLAS

eed to protect system, or eve

Spontaneous

The placenta

Since Mom’s

roblastosis fetveral reasons

If an Rh- wometus. We sha

happens if mo.

Mom can ma

Type of antib

IgG antibodie

Why not usuaaby, so during

Subsequent petus. Effects

P

lasma membr factor). Nam

e this antigen ormed on theis called “D”.

e protein (Rh”.

(Rh-). The imin contact wit

s a simplifica” R factors. Tesponding an

f incompatibilSTOSIS FETA

the developinery pregnancy

s abortion can

is a barrier b

immune syst

talis: the syss this can hap

man reproducll see that the

om is Rh- (the

ke the antibo

ody produced

es can cross t

ally harm firstg the first pre

pregnancies: range from no

rane acting asmed after the with us, and e

em. More tha Two possibil

+). The immu

mmune cells cth a foreign c

tion, as thereThe most comtibody, “anti-D

ity between mALIS

ng fetus from y would end i

n be a result.

between mate

tem is in her b

tem breaks dppen. Rh inco

ces with and Re problem is w

erefore, make

dies, but only

d: IgG (more

the placenta a

t Rh+ fetus? gnancy, the b

level of antibone/little to se

s an “ID flag” Rhesus early n one variantlities:

une cells

can make ell containing

e are 5 mmon is calledD”.

mother and

Mom’s n a rejection!

ernal blood &

blood stream

own, and Moompatibility =

Rh+ male, ½ owith the secon

es the anti-Rh

y if her blood

detail in Imm

and destroy fe

It takes a whibaby probably

body is higherever, even sp

-

t,

g

d

!

fetal blood.

, this is one w

om’s immune most commo

of the concepnd “positive c

h antibody) an

comes in con

munity Chap

etal red blood

ile to make any will not be a

r from the begpontaneous ab

way the fetus

system attacon reason.

ptions will resconception”.

nd is pregnan

ntact with the

pter)

d cells.

ntibodies thataffected.

ginning. Morbortion.

is protected.

ks the develo

ult in an Rh+

t with a secon

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t can affect th

re damage to

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nd

en.

he

the

Step 2. Common Blood Tests, and the “Coulter Counter Readout” We will be learning about some common blood tests. We will not be preforming most of them in lab. The student should know their names, their abbreviations on a sample blood test “readout” (“Coulter Counter Readout”), and the units involved. The student should be able to analyze a mock “Coulter Counter Readout”, which we are using to represent what one might see in a clinical setting. A) Hematocrit.

Your hematocrit is the percent by volume of your blood that is cellular (as opposed to plasma), technically the percent by volume of RBCs. Males have an average hematocrit of 38-54%, females 36-47%. A males higher hematocrit is due to the average increase in size and muscle mass, and therefor a greater need for oxygen transport. Altitude also greatly affects hematocrit. For high altitude residents: about 45% - 61% in males; 41% - 56% in females (These levels gradually average higher as the altitude where people live increases. This is a result of the increased demand for the oxygen-carrying capacity of red blood cells at higher altitudes where there is decreased oxygen concentration in the atmosphere.) Readings outside of these normal ranges can indicate a blood abnormality. Blood analyzers, such as the Coulter Counter, estimate hematocrits in a fraction of a second. The abbreviation is Hct. An alternative method is to centrifuge blood that has been collected in a capillary tube, and then estimate hematocrit using a "reader" device of some sort. Hematocrits taken by use of a centrifuge are referred to as "spun" hematocrits.

B) Blood

Tw Inpasuif

T Tthanfo T Csytuthbew

Clotting – A

he ability of thwounds. Thes

n lecture, we sathway”, whicummary that you can follo

he intrinsic an

he reactions thromboplastnd a group of

or the interact

he Common

Clotting involveynthesized in urn causes fibhreads that foecome fine en

wound.

A review from

he blood to fose test ARE N

saw that the pch lead to a “cis important tw it on the dia

nd extrinsic p

that lead to ptin, blood plaf chemical comtion of the abo

Pathway: Fo

es two plasmthe liver. To

brinogen to corm the basic nough to trap

m lecture mat

orm a clot proNOT given on

process of blocommon patho understandagram below

athways: For

rothrombin acatelets and chmpounds whiove is describ

orming Fibrin:

a proteins, pr form a clot p

onvert to activmeshwork of

p RBCs, thus f

terial

tects the indivthe Coulter C

ood clotting firhway”. Pleaseding clotting te:

rming a molec

ctivation invohemicals theyich we will cabed in your tex

rothrombin aprothrombin mve fibrin. Fibrf the clot. Bleforming a plu

Common pathw

vidual from exCounter Read

rst involves ae see the lectests done in a

cule called “P

lve calcium iy contain whicll the accessoxt and will als

and fibrinogemust first activrin is a proteineeding stops wug of fibrin and

way

xcessive bleedout!

and “intrinsic ature notes fora lab. You sh

Prothrombin A

ions, a substch we will callory factors. Aso be discuss

en, both of whvate to thrombn that organizwhen the fibrid RBCs which

eding from mi

and extrinsic r details. Herehould check to

Activator”:

tance called l platelet fact

A possible schsed in lecture.

hich are bin. Thrombinzes into long, n mesh has h blocks the

inor

e is a o see

tors, heme .

n in sticky

Vitamin K is also important for clotting in that it is needed for the synthesis of prothrombin by the liver.

Intravascular clotting within healthy vessels is normally prevented by an anticoagulant called heparin which is present in the plasma. Heparin is thought to be produced by mast cells found in the tissues of various organs.

Neither of these two tests we will study are tests we have current capability to do in lab at MATC. Know their names and significance for the lab practical. Historically, there are a number of different test procedures that have been used to estimate a person's ability to clot. Two of the more sophisticated are:

1. PT Test = Prothrombin Time Test

This test determines the amount of prothrombin in the blood and is a test of the extrinsic clotting pathway and common pathway. It may be used to follow the effects of coumarin, or other Vitamin K inhibitors, since Factor VIII which is part of the extrinsic pathway, but not part of the intrinsic pathway, is most sensitive to Vitamin K.

2. PTT Test = Partial Thromboplastin Time Test

This test is a test of the intrinsic pathway and common pathway. If both PT and PTT test times are prolonged, then the problem is with the common pathway.

C) Other Blood Tests.

On the Coulter Counter Readout, you may also see the normal ranges and tested values for: - Total Red Blood Cell Count (the number of Erythrocytes per cubic mm of blood). See “Step 3 – Formed Element ID” for a discussion, abbreviations and values. - Total White Blood Cell Count (the number of ALL white blood cells per cubic mm of blood). See “Step 3 – Formed Element ID” for a discussion, abbreviations and values. - Differential White Blood Cell Count (the percentages of the individual types of White blood cells). See “Step 3 – Formed Element ID” for a discussion, abbreviations and values. - Hemoglobin (Hgb) - The hemoglobin test is often used to check for anemia or polycethemia, usually along with a hematocrit or as part of a complete blood count (CBC). - Mean Corpuscular Count (MCV) - is a measure of the average volume of a red blood corpuscle. It is HCT/[RBC]. The normal range for MCV is 80–100 fL. This is a test for anemia or polycethemia.

1 femtoliter = 10-15 liter. I will never make the student convert! You do not see this unit very often. - Mean Cell Hemoglobin (MCH), is the average mass of hemoglobin per red blood cell in a sample of blood. This is a test for anemia and polycethemia. - Mean Corpuscular Hemoglobin concentration (MCHC) is the average concentration of hemoglobin in red blood cells. This is a test for anemia and polycethemia. - Coagulation Time – how long it take to coagulate. Do not worry about normal ranges. This can be used to monitor anticoagulation effects, such as high-dose heparin before, during, and shortly after procedures that require intense anticoagulant administration, such as cardiac bypass, cardiac angioplasty, thrombolysis, extra-corporeal membrane oxygenation (ECMO) and continuous dialysis. This test is not on the Coulter Counter Readout.

- (Rarva

Red blood ceRBC) volume re a standardariation in cel

ell distribution that is report

d size of aboul size.

Exam

width (RDW ted as part of t 6-8 μm in di

mple “Coul

is a measurea standard coiameter. Certa

ter Count

e of the rangeomplete blooain disorders

ter Reado

e of variation od count. Usua, however, ca

out”

of red blood cally red bloodause a signific

cell d cells cant

Step 3. - For eacversus Wbloodstr A. ERYT

D

Biconc

The red cIn a bloodErythrocycenter. Un Sometimethose we staining o

Normal shape

Formed El

ch blood ceWBC (as a gream, and th

THROCYTE

Description

ave, annucleate

ells are very d smear, you wtes are withonder the micro

es, they are pdescribed. So

of the smear.

Abnormal Shape due

Abnormal sh(sickle cell a

ement ID.

ell/element tgroup) vershe function

S

Ce

e 4

numerous in will see a lot out a nucleus (oscope, they

piled up like coometimes, thi

e to slide prep

hape due to panemia)

type, know us Platelets

n of each ele

ells/mm3 (ul)

4-6 million

the blood. Usof erythrocyte(annucleate)look like pink

oins. As we sis is due to di

ZoIn!

paration

pathology

its descripts, relative pement.

Durati

1

sually, they mes and, some). Their typicak discs cleare

aw, the red cseases or to

oom !

tion, absolupercentages

ion in bloodstream

00-120 days

measure 6-8 μtimes, some

al shape is thar in the middl

cells can also defective pro

ute amountss of WBCs i

m

Trans

μm (micrometisolated leuko

at of a cake de.

have differenocess of prepa

s of RBCs n the

Function

sport O2 and CO

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nt shapes fromaration and

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m

B. PLAT

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D

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Unlike redstained th(reniform) Usually, th Leukocyte

TELETS Description

agments containgranules

are not living s involved in begakaryocytes

small sized dlor and are da

KOCYTES -

Description

rical, nucleated

d cells, leukoche smear. The).

he shape of t

es are divided

Ce

ing 150,

cells. Insteadblood clottings.

iskettes abouarker than red

WBCs

Ce

5is a T

(all of them

cytes have a ne nucleus of th

he nucleus of

d into granulo

ells/mm3 (ul)

,000 – 400,000

d, they are sm. They arise

ut 3μm in diamd cells.

ells/mm3 (ul)

,000-10,000 otal WBC Countm counted togeth

nucleus. It is hese cells ca

f various kind

ocytes and ag

Durati

mall sacs filledfrom big leuk

meter. They a

Durati

her)

de

easily visible n show multip

of leukocytes

ranulocyte (ly

ion in bloodstream

5-10 days

d with kocytes

appear a

ion in bloodstream

epends on type

under the miple lobes, or b

s is different,

ymphoid) cells

m

hemos

m

Im

icroscope, bube indented o

and can be u

s.

Function

stasis (blood clott

Function

mmunity/Defense

ut only after haor “kidney-sha

used to ID the

ting)

aving aped”

e cell.

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ation of WBC

me from the ke typical coed in a little well. They diAs we have

hil, eosinoph

TROPHIL Gr

Description

multi-lobed nucleth 4-5 lobes

trophil are thr of 12-15 μmnto 3 - 5 lobeplasm is trannk colored, sut that nucle

hilia is the cohils, indicatin

Cs:

(i) GRANU

marrow bonolors which hmasses or listinguish th

e said, there il, basophil.

ranulocytes

eus 40

he most comm. You can res connectensparent beso you may us is VERY

ondition of hg a possible

ULOCYTES:

ne. Their cytohelp their recobes. In the emselves byare three ty

Relative %

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mmon leukocrecognize thed by a fine ncause its gramistake it fotypical.

aving too mae bacterial in

3 types

oplasm is riccognition. Th

blood, therey having a lepes of granu

Durati

few

cytes. They hem as their nuclear strananules are s

or an “agranu

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ch in granulehe nucleus ise are immatuess segmentulocyte:

ion in bloodstream

w days, at best

have a nucleus is nd or filamensmall and ulocyte” at

es s ure ted

m

Pha

nt.

Function

agocytize bacteri

a

2. EOSI

The eosithe neutr Generallyfull of grathe neutrvisible. Eosinophmore tha 3. BASO

D

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the nucleu

Basophilsmall: 9- Cytoplasnucleus iof granul Basophilconditionbasophils

De

Bilobed distinc

NOPHIL Gra

nophils are qrophils.

y their nucleanules whichrophil, the nu

hilia is the coan expected:

OPHIL Granu

Description

ucleus, but so maning granules thas usually not visi

s are the rar10 μm in dia

m is very ricis bi- or tri-loes which hid

ia is the n of too manys:

escription

nucleus, very ctive staining

anulocytes

quite rare in

eus is bi-lobeh assume a ucleus is stil

ondition of h

ulocytes

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ch in granuleobed, but it isde it.

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ed and kidnecharacteristl easily

aving

ferential count

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they are often se%, as many peo

ave allergies

tes: less tha

es which takes hard to see

erential count

to 4% ot quite the rares

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They have th

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Durati

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few

an 1 %. They

e a dark pure because o

Duratio

st, nd

few

he same size

The cytoplasge color. As

ion in bloodstream

w days, at best

y are quite

rple color. Thof the numbe

on in bloodstream

days, at best

e as

sm is s with

m

Relehepa

he er

Destroyinvolve

during an

Function

ase histamine anarin, among othe

chemicals.

Function

y parasitic wormed in inflammation allergic respon

nd er

s, on nse.

BecauseThey havcells: lymwhereas granuloc

Cell N

Agranulo

1. Lymp

2. Mon

1. LYMPLymphocdiameterthey are The cytocomparis One type 2. MONO Monocytereniform The cytoglass". Monocyte

e usually thesve a compac

mphocytes anlymphocyteytes.

Name

ocytes:

hocytes nuc

ocytes n

PHOCYTES cytes are qur and generastill a few la

plasm is transon to the ce

e makes anti

OCYTES

es are the bor horseshoplasm is tran

es become m

(ii) AGRAN

se cells appct nucleus annd monocyte

es spring from

Description

leus usual spher

nucleus U-shaped

ite common ally they are rger than re

nsparent. Thell and it occ

ibodies (T-ly

iggest leukooe-shaped nnsparent, bu

macrophage

NULOCYTES

ear lacking ind a transpaes. Their loom lymphatic

Different

rical 2 to

d 20 to

in the bloodsmaller thand cells.

he nucleus iscupies most o

ymphocytes)

ocytes: 16-20ucleus, in so

ut with an ap

es during ch

S (or “lymph

in granules, arent cytoplaok is similar,

organs, mo

tial count Du

o 8%

o 40%

d: 20-40%, 8n the other le

s round and of it.

).

0 μm. They home cases eppearance of

ronic infectio

hoid” cells): 2

they are alsasm. There abut their ori

onocytes hav

uration in bloodstr

hours to years

months

8-10 μm in eukocytes b

large in

have a greaeven bi-lobedf "ground

ons.

2 types

so named agare two typegin is differe

ve the same

ream

! T-ly

B-l

Turn induring

but

at d.

granulocyteses of lymphoent. In fact, origin as the

Function

ymphocytes makeantibodies

lymphocytes arephagocytes

nto MACROPHAGg chronic infectio

s. id

e

e

e

GES ons

APPPENDIX: Exampple Coulter Couunter Reeadout