Diagnostic and Biomedical Techniques

Presented by group # 03
Ali Hamza (19337)
Saba (19347)
Areeha(19343)
Arij (19311)
Nayab (19351)
Maham (19349)

Presented to: Sir Muhammad Mudassar

Flame photometry
Purpose
 It is the branch of atomic spectroscopy, is used for qualitative
and quantitative analysis of certain metal ions such as `
sodium, potassium, lithium, calcium and cesium` etc.
Principle
 The principle of flame photometer is based on the
measurement of the emitted light intensity when a metal is
introduced into the flame. The wavelength of the colour gives
information about the element and the colour of the flame
gives information about the amount of the element present in
the sample.
Applications:
 Flame photometry has both qualitative and
quantitative applications
 Flame photometer with monochromatic filters emits

radiations of characteristics wavelengths, which helps

to detect the presence of particular metal in the
sample.
 It is used to estimate sodium, potassium, lithium,

calcium and cesium` etc level in biological samples

like, serum, urine, CSF, and other body fluids.
 Sodium and potassium present in heart and other

muscles can be determined by diluting the blood

serum and aspiration into the flame. 
Spectro-photometry
Purpose:
 Spectro-photometry is a branch of electromagnetic
spectroscopy concerned with the quantitative
measurement of the reflection or transmission
properties of a material as a function of wavelength.
This method is used to estimate the level of an
analyte in solution. Spectrophotometry
uses photometers, known as spectrophotometers,
that can measure the intensity of a light beam at
different wavelengths.
Principle
 Its working principle is based on Beer Lambert law
according to which when a monochromatic light of
certain wavelength is passed through a absorbance
medium its intensity decreases exponentially as the
concentration of absorbing medium increases,
provided the length of absorbing medium is
constant.
Applications
Some of the major applications of spectrophotometers include
the following:
Detection of concentration of substances
Detection of impurities
Structure elucidation of organic compounds
Monitoring dissolved oxygen content in freshwater and marine

ecosystems
Characterization of proteins
Detection of functional groups
Respiratory gas analysis in hospitals
Molecular weight determination of compounds
The visible and UV spectrophotometer may be used to identify

classes of compounds in both the pure state and in biological

preparations.
Turbidimetery
Purpose
 This method is used for determining the

concentration of a substance in a solution by

measuring the loss in intensity of a light
beam through a solution that contains
suspended particulate matter.
Principle
 Turbidimetry is the process of measuring the
loss of intensity of transmitted light due to
the scattering effect of particles suspended in
it.
 Light is passed through a filter creating a

light of known wavelength which is then

passed through a cuvette containing a
solution.
Nephlometery
 It is a technique used in immunology to
determine the levels of several blood plasma
proteins.
 For example the total levels of antibodies

isotypes or classes: Immunoglobulin M,

Immunoglobulin G, and Immunoglobulin A.
 It is important in quantification of free light

chains in diseases such as multiple myeloma.

Principle
 If a substance is completely dissolved in
solution then there will be nearly no scattered
light leading to a low nephelometric value.
But if the solubility limit is reached and the
substance starts to precipitate, the light will
be scattered strongly leading to a
high nephelometric value.
Applications
 It is used for determination of total protein in biological
samples like urine, CSF, serum and other body fluids.
 It is used for determination of immunoglobulins like IgG,

IgE, IgA, IgM, in serum and other body fluids.

 It is used for Determination of amylase activity using

starch as substrate.
 It is used for Determination of lipase activity using

triglycrides as substrate.
 It is used for analysis of water for clarity and ions

concentration.
 It is used for Determination of CO2.
 It is used for Determination of inorganic substances like

ammonia, sulphate and barium chloride.

Calorimetry
Principle
 The principle of calorimetry states that for an

insulated system, heat energy lost by the hot

body is equal to the heat energy gained by
the cold body.
 Calorimetric analysis depends on the direct

proportionality between the heat changes

that occur during chemical reactions and the
amount of reacting substances
Applications
 It is used for analysis of protein stability and
folding.
 It is mostly used in protein engineering.
 It is used in antibody domain studies.
 It is used for characterization of membranes,

lipids and nucleic acids.

 It is used for assessment of the effects of

structural changes on molecule stability.