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Experiment No.


Refractory Period,
Extrasystole, and
Compensatory Pause

 To determine the effect of external

stimulation on the turtle heart
Anesthetize the turtle, and expose its heart.
Connect the apex of the heart to the kymograph.

Stimulate the heart with single break shocks.

Allow the heart to resume normal rhythm between
the shocks.

Record the tracings produced on the kymograph.


Kymograph tracing of turtle ventricular contraction

Kymograph tracing of turtle ventricular contraction

with external stimulus applied
Absolute Refractory Period (ARP)
 This is the period covering the upstroke to
around 2/3 of phase 3 of the action
potential (AP).
 No AP is initiated no matter how strong
the stimulus may be.
Relative Refractory Period (RRP)
 This is the period from last 1/3 of phase 3
to the threshold potential.
 If an extra strong stimulus is applied here,
an AP may be induced.
 This action potential is called an
extrasystole, which is followed by a
compensatory pause.
Effective Refractory Period (ERP)
 This is the period covering the ARP and a
little bit longer.
 Stimuli enters the RRP and an AP is
produced, but it is not conducted to the
myocardial muscle.
stronger contraction

normal normal

Stimulus Extrasystole Compensatory Pause

 A premature contraction of the heart
 Occurs as a response to stimulation from
ectopic foci
Ectopic foci – regions of heart that can initiate the heart
beat other than the SA node
Compensatory Pause
 A rest period after extrasystole
 Duration is longer than normal interval
between contractions
 Allows the heart to resume its normal rhythm
 Allows the ventricles to completely refill

 External stimulation of the turtle heart

causes a premature beat (extrasystole)
which is then followed by a prolonged rest
period (compensatory pause), allowing the
heart to return to its normal rhythm
 During what phase of the cardiac cycle did the
extra systoles fall?

 The extra systole was observed during the

relative refractory period of the cardiac cycle. The
relative refractory period is the period wherein the
muscle is difficult to excite but nevertheless, can
be excited
 What was the period of rest?

The period of rest is the compensatory pause
which is the prolonged interval before a new
contraction takes place.

 How does it compare with the normal?

The compensatory pause is usually longer than
the normal interval between contractions.
 What is its value?

 The compensatory pause occurs because the

external stimuli in the apex of the heart did not
disturb the normal rhythm of the SA node. This
allows the heart to resume its normal rhythm. It is
its way to ensure that after an extra systole, it will
be able to return to its normal rhythm and allow
the complete filling of the ventricles.
Experiment No. 16


 Todetermine the effect of auricular and

ventricular block in auricular and
ventricular contractions of a turtle heart
Anesthetize the turtle, and expose its heart.
Connect the auricle and ventricle of the
heart to the kymograph.

Insert the heart between the jaws of the

Gaskell clamp, so that the clamp lies on the
auriculo-ventricular groove of the heart.

Increase the degree of the block

gradually until there is a difference between
the auricular and ventricular beats.

Increase the degree of block

until the ventricle stops. Touch the
ventricle with a pin point.

Record and tabulate all results

and observations.
Table 1. Heart rates of turtle before and during block

Condition Beats / minute

Atrium Ventricle
Normal Rate 59 59

With Gaskell 34 31
(AV block)

 Auricular and ventricular block causes the

ventricle to lose the contraction impulse
from the sinus venosus, forcing the
ventricle to produce its own contraction.
 What is the property of the ventricle seen in the

 Automaticity was demonstrated in the experiment.

This is one important property of the cardiac
muscle which has the ability to initiate its own
beat. In this case, the passage of impulses from
the atria to the ventricles is blocked, thus the
Purkinje fibers in the specialized conduction
system of the ventricles become the pacemakers.
 What does the discrepancy in the rates prove?

 Discrepancy in the heart rates demonstrate the

totality of the block. A complete block forced the
ventricular muscles to produce its own
Experiment No. 18

Records Obtained from the ECG
Heart Rate
The formula to determine the HR:

25mm/s x 60s
HR = no. of small boxes

HR = no. of small boxes
Heart Rate

HR = 1500/21

Heart Rate = 71
Different Intervals
 QT intervals

Lead I Lead II Lead III

Different Intervals
QRS complex

Lead I Lead II Lead III

Different Segments
PR segment

Lead II Lead III

Lead I
Different Segments
ST segment

Lead I Lead II Lead III

Experiment No. 19

Blood Typing

To determine the importance of blood typing

and cross matching prior to blood transfusion.

Preparation of patient’s serum

The area of extraction was sterilized

and about 3 mL of blood was
extracted on a subject.

The blood was allowed to stand and clot for

10 minutesand was then centrifuged
until the serum separated from the clot
Preparation of donor’s red cell suspension
The second subject’s fingertip was sterilized
and then punctured. A drop of blood was directly put in
the test tube containing 1 mL of 0.9% NaCl solution.

The tube was centrifuged until a button of red cell

formed at the button of the tube.
All the fluid was discarded.

Red cell suspension

Two drops of the 1st subject serum was added
on the 2nd subject’s red cell suspension

The tube was centrifuged

The supernatant was observed

Results and Discussion

Subject 1: Type O
Subject 2: Type O

Result: No hemolysis and agglutination

(reddish tint of the supernatant)

The blood of the 1st and 2nd subject are compatible.


1. What is the purpose of doing cross matching before

blood transfusion?
Cross matching is done before blood transfusion to detect
presence of antibodies in the recipient’s serum that could
destroy transfused red cells.
2. What do you mean by major and minor cross matching?
Major Cross matching- Patient’s serum is mixed with the
donor’s red cell
Minor Cross Matching- Donor’s serum is mixed with the
patient’s red cell
3. Give the major problems encountered in typing and
cross matching.

Clerical errors is the most common cause

of transfusion accidents.

A compatible cross matching does not guarantee that the

transfused blood will survive normally in the recipient.

Proper blood typing and cross matching lessens

the possibility of transfusion reaction. It ensures
best possible results of blood transfusion. Thus, it is
important to perform these tests before transfusing
blood to a patient.
Experiment No. 20

Bleeding time
Clotting Time
Clotting time
 Objective:

To determine the clotting time of the subject
and compare it to the normal values.
Prepare the materials (lancet,
Cotton balls, 70% alcohol, slide,
Stopwatch, NON-HEPARINIZED When a fibrin thread is observed
CAPILLARY TUBE extending between the gap of the
broken tube, clotting time is
Select and sterilize the area
to be punctured. Air dry.

Wipe off the first drop of blood

Fill the capillary tube with blood.

At thirty-second intervals, break

the capillary tubing into small
Sterilize the site At half-minute intervals,
to be punctured draw the pointed part
and allow to dry. of the lancet across the
drop of blood.

Make a firm quick stab When fibrin threads cling to the

on the site of puncture. end of the pointed part of the lancet,
stop and record the time.

Wipe off the first drop

of blood with dry cotton.

Squeeze the finger gently

and place a large drop of
blood on slide.
Avoid touching the wound.

SLIDE METHOD 3 minutes and 30 2-6 minutes

CAPILLARY 2-6 minutes.

•used to screen for problems in blood clotting or coagulation mechanism
•This procedure tests the function of the blood vessels in blood clotting.
•Delayed or lack of clotting can be due to a deficiency in one or more
coagulation factors.
Bleeding Time
 Objective:

To determine the bleeding time of the patient,
compare it with the normal value and know
the significance of this test.
Duke’s Method
Prepare the materials (lancet,
filter paper, cotton balls, Repeat every 30 seconds until
70% alcohol, stopwatch) the bleeding stops.

Sterilize the site to be punctured

and allow to dry.

Make a firm quick stab on the site

to ensure free flow of blood.

At thirty seconds interval, blot

blood coming out of the puncture
by means of filter paper.
 Bleeding time is the time that elapsed between the appearance of
the blood and the cessation of bleeding.

 Bleeding time depends on the rate at which a stable platelet

thrombus is formed and thus, measures the efficiency of the
vascular and platelet phases.

 Bleeding time is principally a measure of platelet plug formation.


 elasticity of cut tissue
 ability of the blood vessels to constrict and retract
 mechanical and chemical action of platelets in the formation of platelet