KUMASI TECHNICAL UNIVERSITY

DEPARTMENT OF LABORATORY TECHNOLOGY

END OF FIRST SEMESTER EXAMINATION FOR 2017/2018 ACADEMIC YEAR
COURSE: BIOPHYSICS CODE: MLT 111
PROGRAMME: MEDICAL LABORATOEY TECHNOLOGY-HND I TIME: 3 HRS
ANSWER QUESTION 1,2,3 AND CHOOSE ANY OF QUESTION 4 OR 5
1.Distinguish with objective lens and eyepiece lens in diagram, the difference between a Microscope and
a Telescope. (4 Marks)
ii. Define the following:
i. Reflection ii. Refraction iii. Diffraction iv. Interference (4 Marks)
c. State the following:
(a) The laws of Reflection (b) The laws of Refraction (c) A Real image
(d) A Virtual image (e) Rectilinear propagation of light (5 Marks)

d. i. What is a Critical Angle? (3 Marks)

ii. State two (2) properties of an image in the following:
I) Concave lens II) Convex lens III) Mirror (6 Marks)
e. Distinguish between the following:

i. A Maser and a Laser (2 Marks)

ii. A visible light, an infra-red and an ultra-violet radiation. (3 Marks)

f. i. A 2 cm high diamond ring placed 30.0 cm from a concave mirror with radius of curvature 45.0 cm.
Determine:
(a) Position of the image (b) Size (4 Marks)
ii. Calculate the speed of light in diamond.
[Take refractive index of diamond to be 2.42] (2 Marks)

g. A flashlight beam hits a surface of a plane glass (n=1.58) at a 70° angle to the normal. What is the
angle of Refraction? (3 Marks)

2. Explain briefly the structure of an Atom and the concept of Coulomb’s law in charges.
(4 Marks)
b. i. Explain the following: A conductor, a semi-conductor, an insulator, electric field, electric current,
direct current and alternating current. (7 Marks)
ii. A tungsten filament of coefficient of resistance of 0.0045°𝐶 −1 used in a flashlight bulb operates at 0.20
A and 5.4 V. If its resistance at 30°C is 1.5 Ω, what is the temperature of the filament when the flashlight
is on? (3 Marks)

3. a. Explain the concept of electrochemistry (2 Marks)

b. Define Oxidation reaction and Reduction reaction. Which reaction occurs at the anode and which at the
cathode? (4 Marks)

c. One-half of an electrochemical cell consists of a pure nickel electrode in a solution of Ni 2+ ions, the
other half is a cadmium electrode immersed in a Cd2+ solution. The cadmium electrode is oxidized and
Nickel reduced. The half-cell potential for Cd and Ni on the emf series table in their reduction states are -
0.403 V and -0.250 V, respectively. If the cell is a standard one, (i) Write the half-cell reactions and the
spontaneous overall reaction. (ii) Calculate the voltage that is generated. (6 Marks)
d.i. What mass (in grams) of nickel could be electroplated from a solution of nickel (II) chloride by a
current of 0.25 amperes flowing for 10 hours? (4 Marks)

ii. Find the time required to deposit 1.92 g of copper from CuSO4 solution by passing a current of 1.5
ampere. (4 Marks)

e. Briefly describe the operation of any one (1) device application that uses the principle of
electrochemistry in a biochemistry lab. (5 Marks)

4. What is meant by electrophoresis? (2 Marks)

b. What is the main function of electrophoresis? (3 Marks)

c. The gel matrixes widely used in electrophoresis are mainly? (3 Marks)

d. Describe the apparatus needed for conducting gel electrophoresis in a biochemistry laboratory
(10 Marks)

e. Mention the correct type of electrophoresis for the following:

i. Gel has a pH gradient

ii. Separate large DNA fragments

iii. Separate protein without SDS

iv. Separate proteins based on electrophoresis and reaction with antibodies

v. Separate ionic species by charge, friction force and hydrodynamic radius

(7 Marks)

5. What is sedimentation and what basic principle is involved in the sedimentation process?
(3 Marks)
b. Define the erythrocyte sedimentation rate (ESR). Mention three (3) causes each of elevated and
reduced ESR. (5 Marks)
C. Explain the processes involved in terms of sample, equipment, method, required value and precautions
used in the Westergren method for measuring ESR. (10 Marks)
d. Discuss four (4) factors affecting red cell sedimentation. (4 Marks)
e. What are some of the significance of reticulocyte count? (3 Marks)

EXAMINER: Mr. Isaac Amankwaah

 MARKING SCHEME

1. Microscope is used for magnifying small objects that are at a short distance from the viewer
whiles Telescope is used for magnifying large objects that are at a large distance from the viewer.

TELESCOPE MICROSCOPE

Eyepiece lens

Objective lens Eyepiece lens Objective lens

(4 marks)

B. ii..i. Reflection is the bouncing back of light rays when it meet an obstacle

ii. Refraction is the bending of light rays when it travels from a denser medium to a less dense medium
with different optical density.

iii Diffraction is the spreading of light rays when it passes through a narrow slit or aperture.

iv. Interference is a phenomenon in which two or more waves superimpose to form a resultant wave of
greater, lower or the same amplitude.

(4 marks)

c. a. The laws of Reflection:

1. The incident ray, the reflected ray and the normal at a point of incidence all lies on the same plane.

2. The angle of incidence is equal to the angle of reflection

b. The laws of Refraction:

1. The incident ray, the refracted ray and the normal at a point of incidence all lies on the same plane.

2. The ratio of the sine of the angle of incidence to the sine of the angle of refraction is constant for any
two given pair of medium.

c. A real image is formed when the actual rays converge and meet at a point which is form on a screen

d. A virtual image is formed when the apparent rays converge and meet at a point.

e. Rectilinear propagation of light states that light travels on a straight line.

(5marks)

d.i. Critical angle is an angle formed when light rays move from dense medium to less dense medium at
an angle of 90°. (3 Marks)
ii. I. Concave lens: Images are; 1. Diminished 2. Virtual

II. Convex lens: Images are; 1. Magnified 2. Real

III. Mirror: Images are; 1. Laterally inverted 2. The same size/distance as the object

(6 marks)

e.i. MASER: Microwave Amplification by Stimulated Emission of Radiation

LASER: Light Amplification by Stimulated Emission of Radiation

ii A visible light: An electromagnetic radiation that can produce a visual sensation

An infra-red radiation: is emission of energy as electromagnetic waves in the portion of the spectrum just
beyond the limit of the red portion of visible radiation. The wavelengths of infrared radiation are shorter
than those of radio waves and longer than those of light waves.

Ultra-violet radiation: Radiation lying in the ultraviolet range; wavelengths shorter than light but longer
than X rays. (5 marks)

f.i.a. 1/f = 1/du + 1/dv, where f= focal length, du= object distance , dv= image distance , r=2f

1/dv = 1/f – 1/du , 1/dv = 1/22.5 – 1/30 = 30dv= 22.5dv + 675, 30dv - 22.5dv = 675. 7.5dv = 675.
dv = 90.0cm. The image is 90.0cm away from the mirror. (2 marks)

b. m = hv/hu, where m= magnification, hu= object height, hv= image height, m= 90/30= 3.00 so the size
of the image is hv = m × hu, v = 3 × 2 cm= 6 cm (2 marks)

ii. 𝑛 =𝑐𝑣 , v = c/n , where n = refractive index of a diamond, c= speed of light in air and v= speed of light
3
in the medium. 𝑣 = 2.42×108 = 1.24×108m/s (2 marks)

g. ni ×sini=nr ×sinr ; (1)sin70° = (1.58)sinr = r = 36.5° (3 marks)

Q2.a.i. An Atom is the smallest particle of an element that can be taken part in chemical reaction. An
atom can be splitted into smaller particles including protons, electrons and neutrons.

Coulomb’s law of electrostatics states that the force between two charges is directly proportional to the
products of the charges and inversely proportional to the square of their distance between the charges.
𝐹 = 𝑘 × 𝑄1 × 𝑄2⁄ 2 (4 marks)
𝑟
b.i A conductor is a substance that allows an electric current or heat energy to flow through them.
A semi-conductor is any material that has the ability to permit or allow the flow of charges through them
partially thus they behave both as a conductor and an insulator.
An insulator is a material that does not allow charges to flow through them.
Electric field is an area or region where electric force is felt or experienced.
Direct current is s current that flows in one direction.
Alternating current is a current that can move sinusoidal or flow forward and backwards.
(7 marks)
ii. The resistance of the filament when the flashlight is on R = V / I = 5.4 V / 0.20 A = 27 Ω
PT=PO(1+α(T-TO)) T=TO+1/α(R/RO-1)
30°C+( 1 / 0.0045°𝐶 −1)(27 Ω/1.5 Ω - 1)= 3807.78 °C (3 marks)

Q3
a. Electrochemistry is the study of interchange between chemical energy and electrical
energy. Many significant chemical reactions are electrochemical in nature.
Electrochemical reactions are chemical reactions in which electrons are transferred
(2 marks)
b. Oxidation reactions: atoms of the element(s) involved in the reaction lose electrons. The
charge on these atoms must then become more positive.
Eg: Fe2+ (aq) = Fe3+ (aq) +e-
Whiles in reduction reactions, atoms of the elements involved gain electrons.
Eg: Zn2+ (aq)+ 2e-= Zn(s)
Oxidation = Anode,
Reduction = Cathode (4 marks)
c. the cadmium electrode will be oxidized and nickel reduced because cadmium is lower in
the emf series; thus the spontaneous reactions will be
Cd = Cd2+ +2e-
Ni2+ + 2e- = Ni
-------------------------------------------------
Ni2+ + Cd = Ni + Cd2+ (3 marks)
From the standard emf series table, the half-cell potentials for cadmium and nickel a
are, respectively -0.403 V and -0.250 V.
Therefore, ΔV = V°Ni -V°Cd = -0.250 V – (-0.403 V) = -0.153 V (3 marks)
d. i. m = (M× I× t) / (n× F) where m = mass of substance, M = molecular mass, I= current
passed in ampere, t= tine in seconds, n= number of electrons transferred, F= Faraday’s
constant (96475 C/eqv).
Nickel undergoes reaction according to Ni2+ + 2e- = Ni, therefore
m= (58 g / mol × 0.25 A × 36000 s) / (2e × 96475) = 2.71g (4 marks)

ii. m = M I t / n F where m = mass of substance, M = molecular mass, I= current passed

in ampere, t= tine in seconds, n= number of electrons transferred, F= Faraday’s constant
(96475 C/eqv).
CuSO4 = Cu2+ + SO42-
Cu undergoes reaction according to Cu2+ + 2e = Cu
Therefore t= (m× n× F) / (M× I) = (1.92 g × 2e × 96475 C) / (1.5 A × 64 g/mol) = 3860s
 (4 marks)

e. ION SELECTIVE ELECTRODES: (I.S.E.) Principle

An ideal I.S.E. consists of a thin membrane across which only the ion to be measured can
be transported. The transport of ions from a high concentration to a low one through a
selective binding with some sites within the membrane creates a potential difference.
This forms the basis of ion selective electrode.
OR
pH Electrodes
Principle: The H+ ions either diffuse out or into the gel layer of the glass membrane
depending on the pH value of the measured solution. Since the glass electrode has an
internal buffer with a constant pH value, the potential at the inner surface of the
membrane is constant during the measurement. The total membrane potential is a of the
difference between the inner and outer charge. From the potential, the H+ ions
concentration can be determined using Nernst equation.
OR
pO2 (Clarks) Electrode
Principle: Biosensors are analytical instruments that convert biochemical signal into
quantifiable electrical signal.
OR
Electrochemical Detectors
Introduction
Electrochemical detection (ECD) technique used for High Performance Liquid
chromatography (HPLC) is an extremely selective detection technique.
ANY ONE FOR (5 marks)

Q4

a. The movement of particles under spatially uniform electric field in a fluid is called
electrophoresis. Electrophoresis of positively charges particles (cations) is called cataphoresis.
Electrophoresis of negatively charges particles (anions) is called anaphoresis. (2 marks)
b. Electrophoresis technique applies a negative charge so proteins move towards a positive charge.
This is used example for the separation of deoxyribonucleic acid (DNA), ribonucleic acid (RNA),
or protein molecules using an electric field applied to a gel matrix. (2 marks)
The rate of migration of particle depends on the strength of the field, on the net charge size and
shape of the molecules and also on the ionic strength, viscosity and temperature of medium in
which the molecules are moving. (1 marks)
c. Gel matrix used mainly is mainly polyacrylamide and agarose. (3 marks)
d. The equipment and supplies necessary for conducting agarose gel electrophoresis are relatively
simple and include:
1. An electrophoresis chamber and power supply.
2. Gel casting trays, which are available in a variety of sizes and composed of UV transparent
plastic. The open ends of the trays are closed with tape while the Gel is being cast, then
removed prior to electrophoresis.
 3. Sample combs, around which molten agarose is poured to form sample wells in the gel.
4. Electrophoresis buffer, usually Tris-acetate-EDTA (TAE) or Tris-borate- EDTA (TBE).
5. Loading buffer, which contains something dense (eg. glycerol) to allow the sample to “fall”
into the sample wells, and one or two tracking dyes, which migrate in the gel and allow visual
monitoring or how far the electrophoresis has proceeded.
6. Ethidium bromide, a fluorescent dye used for staining nucleic acids.
NOTE: Ethidium bromide is a known mutagen and should be handled as a hazardous
chemical – wear gloves whiles gloves while handling.
7. Transilluminator (an ultraviolet light box), which is used to visualize Ethidium bromide
stained DNA in gels. NOTE: always wear protective eyewear when observing DNA on a
transilluminator to prevent damage to the eyes from UV light.
Any five for two marks each(10 marks)

e. i. Gel has a pH gradient = electrofocusing gels. (1 mark)

ii. Separate large DNA fragents = Agarose gels (2 marks)
iii. Separate protein without SDS = Native gels (1 mark)
iv. Separate proteins based on electrophoresis and reaction with antibodies =
Immunoelectrophoresis (1 mark)
v. Separate ionic species by charge, friction force and hydrodynamics radius = capillary
electrophoresis. (2 marks)

Q5
a. Sedimentation also known as settling, may be defined as the removal of solid particles from a
suspension to form separate layers called scum and sludge. (2 marks)
The basic principle involved in the sedimentation process is force of gravity.
(1 marks)
b. Erythrocyte Sedimentation Rate (ESR) is define as the measurement after 1 hour of the
sedimentation of red cells when blood is allowed to stand in an open ended glass tube mounted
vertically on a stand. (2 marks)
Causes of elevated ESR
1. Tuberclosis , 2. Rheumatoid arthritis, 3. Myocardial infarction, 4. Pregnancy,
5. Multiple myeloma and other plasma cell disorders, 6. Neoplastic diseases
7. Degenerative diseases ANY THREE (3) FOR 1.5 MARKS
Causes of reduced ESR
1. Polycythaemia, 2. Hypofibrinogenemia, 3. Hereditary spherocytosis,
4. Congestive heart failure, 5. Sickle cell anemia ANY THREE (3) FOR 1.5 MARKS
c. Western method
The Western pipette is a glass pipette 30 cm in length and 2.5 mm in diameter. The bore is
uniform to within 5 % throughout. A graduated scale in mm extends over the lower 20 cm.
Sample: Venous blood collected in EDTA. Four volumes of blood is diluted in 1 volume of
citrate. Alternatively, 2ml blood is directly collected in 0.5 ml of 3.8 % trisodium citrate.
 (1 mark)
Equipment required
1. Western pipette
2. Stainless steel rack for holding pipette
3. Timer
Method
1. Mix the blood sample well and draw in the Westergren pipette to the 0mm mark with a
rubber teat.
2. Place the tube exactly vertical in the rack which has a rubber cork at the bottom. Fix with
adjustable screws after removing the teat.
3. Leave undisturbed for exactly 60 min free from vibrations and not exposed to direct sunlight.
4. Read the column of clear plasma above the column of sedimented red cells to the nearest mm.
Normal value; Males: 0-10 mm 1st hour Females 10-20 mm 1st hour (1 mark)

Precautions
1. Westergren pipette must be clean and dry.
2. Dilute the blood sample just before setting up the ESR.
3. Never use mouth suction to fill the pipette.
4. The Westergren pipette must be placed vertically.
5. The ESR must be set up at room temperature (18-25°C) (2 marks)
d. FACTORS AFFECTING RED CELLS SEDIMENTATION
The rate of fall of red cells is influenced by a number of factors which interact with interest with
each other closely. These are
1. Rate of rouleaux formation: Sedimentation of red cells is greatly influenced by the extent to
which the red cells form rouleaux. More is the formation of rouleaux, higher will be the ESR.
Rouleaux formation is in turn influenced by the concentration of fibrinogen and other acute
phase proteins such as CRP, haptoglobin, ceruplasmin and others. Rouleaux formation and
thus increase ESR. On the other hand, albumen retards rouleaux formation and ESR.
2. Ratio of red cells to plasma or the hematocrit also influences ESR. In anemic patients, the
number of red cell is less and the proportion of plasma is more. The higher proportion of
plasma increases rouleaux formation and hence increases ESR.
3. Type of red cells sickle cells and spherocytes do not form rouleaux and hence ESR is
decreased in hereditary spherocytosis and sickle cell anaemia.
4. Plasma viscosity ESR increases as the plasma viscosity increases. Viscosity is dependent on
the concentration of fibrinogen and other plasma proteins.
5. Length of the tube: Longer the tube, longer will be the duration of the second phase and
higher will be the ESR.
6. Viscosity of the tube: If the tube is not vertical, sedimentation occurs more quickly due to
streaming of blood down the wall of the sloped tube.
7. Temperature: Sedimentation is accelerated as the temperature increases, ESR is hence
always at room temperature (18-25°C) ANY FOUR (4) FOR (4 marks)
e. Reticulocyte count is a reflection of erythropoietic as in hemolytic anemias. The counts correlate
with the severity of hemolysis. (2 marks)
The count is increased in conditions with accelerated erythropoiesis as in hemolytic anemias. The
counts correlate with the severity of hemolysis.
In nutritional anemias, the count is low. On treatment with hematinics the count rises. Hence,
reticulocyte count is low in aplastic anemia and red cell aplasia. When aplastic crisis occurs in
hemolytic anemia the reticulocyte count is low. (1 mark)