PLETHYSMOGRAPH

Identification:

Aim:
To detect changes in box pressure in combination with either changes of mouth pressure or with flow rate
under defined breathing conditions.

Principle:
1. Based on Boyle’s law, “For a fixed amount of gas in a closed compartment the relative changes in the
compartment’s volume are always equal in magnitude but opposite in sign to the relative changes in
pressure - Pressure x volume = constant or Volume α 1/P.
2. Assumes temperature remains constant, When subject breathes in and out against a shutter, changes in
pressure and volume occur. The animal makes respiratory efforts against the closed shutter, causing their
chest volume to expand and decompressing the air in their lungs.
3. Increase in chest volume slightly reduces the box volume and thus slightly increases the pressure in the
box. These signals are evaluated in order to determine static lung volumes and airflow resistance.

Applications in preclinical studies:

1. To measure respiratory parameters in conscious unrestrained subjects, including quantification
of bronchoconstriction.
2. In anesthetized plethysmography, lung resistance and dynamic compliance are measured directly because
the subject is anesthetized.
3. The double-chamber plethysmograph (dcp) measures respiratory parameters in a conscious restrained
subject, including airway resistance and conductance.

Applications in clinical practice:

1. Body plethysmography - To estimate FRC and RV in obstructive airway diseases.
2. Impedance plethysmography - is a non-invasive method used to detect venous thrombosis in these areas of
the body.
3. Water / air plethysmography - to determine circulatory capacity of arms or legs.
 ELECTROCONVULSIOMETER
Identification:

Principle:
The maximal electroshock (MES) model is a model for grand mal epilepsy and the end point considered as
tonic hind limb extension (THLE) which are evoked by electric stimuli. The aim is to demonstrate the anti-
convulsant activity of screened drugs against maximal electroshock (MES) induced convulsions in rats.
Materials:
Animal/species : Rat/ Wistar
Sex/Body weight : Male/ 150-250 g
Syringe/needle : 1ml/ preferably 23G
Drug : Phenytoin (20-25mg/kg, i.p or po)
Stimulus:
For rat: Intensity of stimulus: (150mA, 50Hz for 0.2sec)
For mouse: Intensity of stimulus: (12mA, 50 Hz for 0.2 sec)
Procedure:
Mouse/rat should be screened 1 week prior to the experiment. The animal present with the positive THLE is
included in the study. One week time is given to animal to recover from the excitatory neuronal discharge in
the brain.
After 1 week, animals are divided into 2 groups,
Group 1: Control group (n = 6); rats are given the saline as per body weight
Group 2: Treatment group (n = 6); rats are given test drug, i.p., thereafter 30 min rats are given electroshock at
the intensity of 150 mA, 50 Hz for 0.2 sec. (in case of oral test drug, MES induced after 60 min).
Seizure score:
0 = no seizure
1 = forelimb extension without hind limb extension
2 = complete forelimb extension and partial hind limb extension
3 = complete tonic hind limb extension (THLE) (hind limb become parallel to the tail)
4 = post ictal depression.
Observations and Results
Percentage protection (%) = No. of animals with THLE absent / total no. of animal × 100.
 MAMMALIAN HEART PERFUSION ASSEMBLY
(LANGENDORFF’S APPARATUS)
Identification:

Principle:
Oscar Langendorff established this procedure in 1897. This is based on the principle of retrograde flow in the
aorta either at constant flow or constant pressure. The entire perfusate enters the coronary arteries via the ostia
at the aortic root. The physiological salt solution (PSS) passes through the coronary circulation providing
constant oxygen and nutrients to the heart and then, perfusate drains into the right atrium via the coronary
sinus. Coronary flow rate is measured on volumetric determination.

Animals required:
1) Albino Rats (300 gm and at least of 1 yr of age),
2) New Zealand Rabbits (1.5-3 kg and 3 years of age)
3) Guinea pig (300-450 gm and 2-3 years of age)

Procedure:
1. Animal is pretreated with the heparin (55-110 µg/ kg, i.p or s.c [for rabbit] or heparin (500–1000 IU/kg,
i.v in rabbit and rat and i.p in guinea pig). Animal is stunned and the thorax is opened immediately and
the heart exposed. Heart is dissected out and removed as rapidly as possible and transferred immediately
into cold Krebs & Henseleit solution (rat and guinea pig heart) or McEwens solution (rabbit heart).
Transfer of isolated heart immediately to the ice cold PSS leads to reduce the metabolic activity as well
as reduce beating, which results into low requirement of glucose and oxygen
2. Cannula is inserted into aorta, but should not cross aortic valve. PSS is perfused at the rate of 20mL/min
(rabbit) or 7-9mL/min (mouse). Maintain PSS flow adequately to prevent edema in cardiac tissues due to
high perfusion pressure.
3. Attach a heart clip complete with a length of thread to the tip of the ventricles; The thread is passed
around a pulley arm vertically below the heart. Attach the thread to the strain gauge transducer after
passing it over a pulley and about 20 cm apart and then the transducer is attached to the coupler of
student physiograph. The recording is done by attaching the thread to a strain gauge transducer, an
attachment with the student physiograph.
4. After the experiment animals are disposed in the yellow polythene bags for the incineration.

Drugs:
Drug added by attaching a syringe to the injection port (or into a rubber tubing) preferably keeping the
volumes < 0.2 ml. Initially the experiment is started with the stimulant drugs (epinephrine, norepinephrine,
isoprenaline and calcium in increasing doses), followed with depressant (acetylcholine, potassium) drugs and
finally antagonists (propranolol and atropine) are added as the effects of the antagonists take a long time to
wear off.
Recording:
Reading can be taken over a period of 30 seconds. If heart movements are too fast to count, kymograph can be
used.

Observations:
The preparation is observed for
1) Inotropic (by measuring the amplitude of contractions) effect
2) Chronotropic (by measuring the rate of heart beat)
3) Effect on coronary vascular resistance (by measuring the rate of collection of drops from the
preparation).
4) Effect of drugs on the isolated heart.

Applications:
1. To study normal cardiac physiology - contractility, heart rate & electrical activity of the heart.
2. To study pathological effects on cardiac tissues. Eg - effects of hypoxia, ischemia, ischemic reperfusion
injury, arrhythmias, etc.
3. Understanding of cell based therapy and cardiac transplantation.
4. To study pharmacological and toxicological effects of drugs on heart.
 STUDENTS PHYSIOGRAPH
Identification:

Principle:
Physiograph is an electronic stimulator which is sensitive enough to convert the mechanical energy to the
electrically amplified impulse to record the response.

Standard transducer for student physiograph:

A transducer is an electronic device which is prompted by energy from one system (mechanical) and converts
energy to another. There are several types of transducers such as pressure transducer, respiration belt
transducer, force transducer, pulse transducer, isotonic transducer, etc. which are very commonly used in the
biomedical research.

Coupler:
1. Coupler is the main device which is attached to the transducer, which should be compatible with
 transducer and controls the input impulse. There are several types of couplers,
2. Strain gauge coupler - mainly involved in the experiment of arterial and venous pulse, blood pressure
recording, BMR, spirometry experiment, in vitro frog heart preparation, isolated tissues, finger
movement, etc.
3. Biopotential coupler - used in the recording of ECG, EEG, EMG, etc.
4. Other less commonly used are pulse/respiration coupler, temperature coupler, etc.

Applications:
1) Recording in cardiovascular physiology (arterial and venous pulse, blood pressure in dog/rabbit, ECG
clinical, effect of vagal stimulation and drugs on perfused isolated frog heart, etc.)
2) Neuromuscular physiology (muscle twist, strength of stimulus, fatigue, isometric contraction, etc.)
3) Experimental respiratory analysis (respiratory movement, lung volumes, tidal volume, IRV, ERV, VC,
IC, BMR, etc.),
4) Human experiment on ECG clinical, EMG, EEG, etc. and to see the effect of drugs on other in vitro
experiments like, isolated intestine, isolated uterus, sciatic nerve, etc