LAB EXAM Flashcards
(RBC counting)
what solution do we use and how much should we take from it?
Hayem’s solution
990 microL
(RBC counting)
is the solution hyper/hypotonic?
what does it contain?
what will happen to the RBC?
hypertonic
NaCl, Na2So4, HgCl2
they will shrink
(RBC counting)
how much blood do we need?
10 microL
(RBC counting)
after mixing blood and solution, how much of it should we take?
20 microL
(RBC counting)
which square should we count from and how many squares?
small square
40
(RBC counting)
counted number of RBC in the 40 squares equals to cell number in-
1/100 microL of the diluted suspension
(RBC counting)
how will we know the number of cells in microL of blood?
x100
bcs of the dilution
x100
bcs of the the total number of the squares
(RBC counting)
normal value of RBC in microL?
4.4-5.5 milion cells / 1microL
(WBC counting)
What solution do we use?
what does it contain?
how much of it do we need?
Türk’s
methylene blue dissolves in acetic acid
90 microL
(WBC counting)
What happens to the RBC and why? (when putting inside the solution)
what will happen to the WBC?
RBC will Lysis
solution will stain the nuclei of WBC
(WBC counting)
How much blood do we need?
How much will we take from the blood+solution?
Add 10 microL blood
Take 20 microL
(WBC counting)
What squares do we use?
How many squares should we count?
Large
25
(WBC counting)
Counted number of WBC in the 25 squares equals?
Cell number in 1/10 microL of the diluted suspension
(WBC counting)
What is the normal value?
6000-8000 cells / 1 microL
(Leukocyte differential count on peripheral blood smear)
all steps in the preparation of the blood?(after we put the blood on the slide)
3 min in undiluted May-Grünwald solutiun
1 min in diluted May-Grünwald solutiun
15-20 min in Giemsa solution
wash with water
(Leukocyte differential count on peripheral blood smear)
how much should we count?
100
(Leukocyte differential count on peripheral blood smear)
How do you prepare yourself for the lab?
gloves and lab coat!!! Tell subject to wash hands Prepare equipment Use alcohol to clean finger Prick the finger and throw away needle
(Leukocyte differential count on peripheral blood smear)
Giemsa solution, what does it made of?
what is it for?
phosphate buffer (pH=6.8) eosinophilic staining
(Leukocyte differential count on peripheral blood smear)
must do before using the slide on the microscope?
what do we use in the microscope to whatch it?
put a drop of immersion oil
the black magnification (special for immersion oil)
(Leukocyte differential count on peripheral blood smear)
how much should you count?
what are the normal values?
count 100 Neutrophils 60-70% Lymphocyes 25-30% Monocytes 4-8% Eosinophils 2-4% Basophils 0-1% **mnemonics!!! Never Let Monkeys Eat Bananas!!!**
(Leukocyte differential count on peripheral blood smear)
how can you identify a neutrophil granulocyte?
1.5-2 times larger than RBC
usually more than 2 nuclei
light eosinophilic (pink) cytoplasm
containing granules (but they are unstained)
(Leukocyte differential count on peripheral blood smear)
how can you identify a lymphocyte?
same size as RBC or bigger big basophilic (purple) nucleus
(Leukocyte differential count on peripheral blood smear)
how can you identify a monocyte?
2-3 times bigger than RBC larges WBC "horeseshoe"/kidney shape eccentric nucleus basophilic (grey-purple) cytoplasm
(ABO type by “one-sided”)
what type of solution should you firstly put on the plate, and where?
seline
under each one- A, B, AB, control
(ABO type by “one-sided”)
what is antigene and where is it located?
it is located on the surface of the RBC, it is the “ID” of the blood type. if i have antigene A i have blood type A.
(ABO type by “one-sided”)
what is antibody and where is it located?
Immunoglobulin (Ig) used in the immune system to help protect cell and so on
they are circulated in the plasma- so if i have blood type A it will have antibodies for B.
(ABO type by “one-sided”)
what type of Immunoglobulin (Ig) are attacking the RBC antigene?
IgM (pentamere)
(ABO type by “one-sided”)
the blood drop which was labeled AntiA got agglutanated- what type is this blood and how do we know that?
the blood is type A.
person with blood type A wont have in his plasma antibodies for A- BCS IF HE DID IT WILL ATTACK THE RBC
(Rh test)
what type of Ig is the antibody for Rh?
anti D= IgG (monomere/dimere)
(Rh test)
what will we observe in case of Rh- person?
no agglutanation bcs this person doesnt have Rh antigene on its RBC surface so there is nothing to “attack”
(transport rate on RBC)
we need two control. positive and negative. describe each one
negative - used as blank. (hypotonic solutiol kills RBC!)
100 microL blood with 3mL distilled water
positive- used as 100% absorbance (isotonic solution!)
100 microL blood with 3mL isotonic solution
(transport rate on RBC)
what does the transport rate deoends on? (2)
- permeability of the membrane for the specofoc substance
2. electrochemical gradient
(transport rate on RBC)
can urea pass the RBC memb?
yes, with UT-B (urea transporter)
(transport rate on RBC)
how do glucose pass the RBC memb?
GLUT1 <3
(transport rate on RBC)
how do water enter the RBC?
aquaporin (3 types):
aq1- constitutively in the memb
aq2- collecting tubule, hormonally regulated by Vasopresin
aq3- constitutively in the memb (can also pass glycerol)
**water can also pass (8%) through urea transporter
(transport rate on RBC)
how can ammonium ions enter the cell?
when they convert to ammonia= NH3+H
the proton goes to the HCO3- buffer
and ammonia enters the cell freely
inside cell becomes NH4 again (intracellular becomes alkaline)
(transport rate on RBC)
what is the reason we will have a sudden drop in the absorbance upon adding NaHCO3?
bcs we have the HCO3/Cl exchanger that enhace that enhances the accumulation of Cl and NH4 inside the RBC
(transport rate on RBC)
first step: how much of bllod? how much of solution we need?
100 microL blood
3mL phys seline
and for all the rest same amount
(transport rate on RBC)
what will happen when we put the blood with urea?
urea enters the RBC through UT-B -> cell will swell (osmotically active)
we have a lower optical density bcs of the cells burst!
(transport rate on RBC)
what will happen when we put the blood with glucose?
glucose enter the RBC slower then urea theough the GLUT1
(transport rate on RBC)
what will happen when we put the blood with NH4Cl?
NH4Cl seperates to NH4+ and Cl
NH4 seperates to NH3 and H
NH3 enters the cell freely
and Cl enters the cell throgh HCO3/Cl exchanger
-> acoumulation of NH4Cl inside the cell slowly
(transport rate on RBC)
what will happen when we put the blood with NH4Cl?
and after 1 min add the NaHCO3
enhances the HCO3/Cl exchanger work so the cells swell and burst more rapidly
CO2 diffuses freely when we add NaHCO3
(ABO type by “two-sided”)
which blood should we use?
unknown tube with serum seperated from the RBC (we can see the seperation)
also we have 3 known RBC (A,B,O)
(ABO type by “two-sided”)
what should we put on the antibody plate?
below anti A we put- drop of antibody A + drop of 10% RBC suspension
same for B and AB
in the 4th place we put- drop of 10% RBC suspension + the unknown serum
(ABO type by “two-sided”)
how do you start the experiment?
COAT! GLOVES!
pour the unknown serum to a different tube
mix the remaining serum+RBC
take 3 drops from it with 1mL of seline (this is our 10% RBC suspension)
(ABO type by “two-sided”)
descibe the blood type plate-
we have A, B, O letters
beneath the letter A we put- A known RBC (it has antigene A on its surface!!!)
we put also the unknown serum
repeat for B, O
(acid base parameters)
what is the normal plasma pH?
7.35-7.42
(acid base parameters)
what is the normal PaCO2 ?
40 mmHg
38-42
(acid base parameters)
what is the definition of a buffer solution?
mixture of a weak acid with its conjugated (strong base) salt andthe opposite
(acid base parameters)
write down the Handerson-Haselbach equation
pH= pK + log [(A-)/(HA)]
(acid base parameters)
give examples for extracellular buffers
bicarbonate
phosphate
plasma proteins
(acid base parameters)
give examples for intracellular buffers
Hb
bicarbonate
organic and inorganic phosphate
(acid base parameters)
what is the standart HCO3- ?
23-25 mM
actual should be the same!!!
(acid base parameters)
what is the normal buffer base values?
what does it mean?
44-49 mEq/L
sum of HCO3- and Albumin anions
(acid base parameters)
what is the normal base axis?
+/- 2.5 mEq/L
how much should we add to the paitent if he has alkalosis or acidosis
(acid base parameters)
how do you know if its a respiratory/metabolic alkalosis/metabolic ???
ROME
Respiratory Opposite- the PCO2 will move opposite to the pH!!
Metabolic Equal - the PCO2 stays the same.
**pH WILL TWLL YOU IF ITS ALKALOSIS/ACIDOSIS
(acid base parameters)
what is the difference in measuring standart and aqtual HCO3- from the graph?
standart HCO3- = where the line intersect with HCO3- line
aqtual HCO3- = we take the actual PCO2 point and draw a line in 45 defrees to the marked lines that are on the HCO3 line
(ECG)
einthoven is
polar/bipolar?
bipolar - 2 electrodes
negative and positive electrodes and the leads are measured btw them
right arm - -
(ECG)
QRS normal range
- 06-0.1 sec
0. 5-2 mV
(ECG)
QT normal range
0.4 sec
(ECG)
ST normal range
+/- 1mV
(ECG)
PR normal range
0.12-0.2 sec
(spirometry)
what is the first task?
how should you breath?
static lung-press IVC
4-5 relaxed breath
exhale completely
inhale maximally
(spirometry)
what is the second task?
how should you breath?
dynamic-press FVC
4-5 relaxed breath
inhale maximally
exhale completely
(spirometry)
what is the third task?
how should you breath?
MVV
30 sec of hyperventilation
(spirometry)
normal TV?
500 mL
(spirometry)
normal ERV?
1200 mL
(spirometry)
normal IRV?
3.1 L
(spirometry)
normal VC?
4.8 L
(spirometry)
normal RV?
1.2 L
(spirometry)
normal FRC?
ERV+RV
2.4 L
(spirometry)
normal TLC?
6 L
(spirometry)
what is FEV1 and what is the important ratio we get from it?
Forced Expiratory Volume after 1 sec
FEV/FVC = 0.8 !
(Smooth muscle activity)
What are the two types of intestinal smooth muscke contraction?
Phasic Contraction (Peristalsis) - Originated from Cajal's pacemaker cells. Tonic Contraction (Basal) - Induced by stretch
(Smooth muscle activity)
What parameters are measured and how?
We measure frequency and amplitude of SMCs contractions. Mechanoelectric transducer is attached to the smooth muscle wall.
(Smooth muscle activity)
What solution the intestine is located in?
Tyrode - Isosmotic solution.
For general extracellular fluid in 38 degrees.
(Smooth muscle activity)
What is added to the solution?
Oxygen pump and Glucose
(Smooth muscle activity)
What is the normal contraction frequency?
6 - 9 per Min
(Smooth muscle activity)
What is the first task?
What do we learn from it?
Hypoxia:
Lowers the frequency and amplitude of contractions.
Reversible as long as there is no tissue damage
(Smooth muscle activity)
What is the 2nd task?
What do we learn from it?
Acetyl-Choline: (ascending conc.)
M3 receptors are activated - Gq activates the SMC to have higher Ca and Increase contractillity tone.
Amplitude is higher (frequency same).
(Smooth muscle activity)
What is the 3rd task?
What do we learn from it?
Atropine:
M3 receptors are blocked- inhibits Ach activation causing Ca signal to be unchanged from normal.
Amplitude is the same. same same but different.
(Smooth muscle activity)
What is the 4th task?
What do we learn from it?
Ach- Esterase: Added stigmazan (Neostigmin isoform) - inihibits Ach-Esterase - causing Ach to stay longer in the Post-synaptic position. Higher Contractillity.
(Smooth muscle activity)
What is the 5th task?
What do we learn from it?
cAMP:
PKA is activated - Phosphorylates and Inactivates MLCK (Myosin light chain Kinase). MLC is not activated - No contraction - Amplitude goes down.
(Fish Heart)
What is the special features of the fishs heart?
1) One circulation cycle.
2) Doesnt contain coranary vessels (Straub heart - Nutrients directly from enviroment).
(Fish Heart)
What is the special solution do we put this shitty fish in?
Ringer Lactate solution.
It provides the heart with ATP.
(Fish Heart)
What is the surgical intervention done in order to help us measure the parameters from this shity fish?
It is a decapitated Fish.
So it has no PARA/SYM connections to periphery.
(Fish Heart)
How do we measure contraction?
The fish heart is hanged by a thread and each contractio pulls the thread and the mechanotransducer picks up the pull.
(Fish Heart)
Observations from the 1st experiment?
Temprature - increase will cause higher contraction frequency and force (and vice versa).
(Fish Heart)
Observations from the 2nd experiment?
Extrasystole and compensatory pause - When introduced with electrical stimulus another systole period is shown. (except from when it is in refractory period).
(Fish Heart)
Observations from the 3rd experiment?
Stannuis Ligatures -
1) Seperation of Sinus venosus and atria of the heart, similar to AV Block. Lower HR.
2) Seperation of Atria of the heart and Ventricles, pacemaking is from Ventricles - Even Lower HR.
