Functions of components:

- The light source provide the energy that the sample will modify or attenuated by absorption.
The light is polychromatic.
- A wavelength selector or filters isolate a portion of spectrum emitted by the source and
focuses it on the sample.
- The sample in a suitable container ( e.g cuvet) absorb a fraction of the incident light and
transmits the remainder.
- The light that pass through cuvet and sample strikes the cathode photodetector and
generate an electrical signal.
- The electrical signal is processed electronically.
- The processed signal is electronically coupled to the display unit .
 Reflectometry:
- Measurement of analysis in biologics fluids using reflectometry has been used for decade l.
- They instruments used for this.
- Reflectometer:
- Is filters photometer that measure the quantity of light reflected by the liquid sample that
has been dispensed onto a grainy or fibrous solid support.
- Two clinical application include urine dipstick analysis and dry slide chemical analysis.
 Type of reflectance:
 Specular:
- Specular reflectance occur on a polish surface where the angle incident of the radiant energy
is equal to the angle of reflection.
 Diffuse.
- Diffuse reflectance occur on non polished surface ( i.e a grainy or fibrous surface ).
- A typical reflectance used in clinical laboratory detect only a constant fraction of diffuse
reflected light.
 Component in the reflectometer:
- Tungsten halogen lamb.
- Monochromatic filters .
- LED display .
- Detector.
- Lens
- Diffuse reflected light.
 Fluorimetry.
- Fluorimetry is based on an energy exchange process that occurs when certain compound
absorb electromagnetic became, became excited, and returned to an energy level lower
than or equal to their original level.
- Because some energy is lost before emissions from the excited state by collision with solvent
or others molecules, the wavelength of emitted light is longer then that of excited light.
 Components of fluorimeter and spectrophotofluorometer:
- A light source
- An excitations ( primary ) Monochromator .
- Cuvet
- An emissions ( secondary ) Monochromator .
- A photodetector .
 Components of a fluorimeter:
- Sources
- Primary filter
- Sample holder.
- Secondary filter.
- Photomultiplier tube.
- Readout.
 Nephelometry and turbidimetry:
- Nephelometry is the measure of light scattered in the particulate solution.
- Nephelometry detects light that is scattered at various angles and this scattered light yields
small signals that amplified.
- Three types of light scattered are symmetric if the wavelength of the light is larger than that
of diameter of the particles, forward light scattering if wavelength is smaller than that of
diameter particle, more light scatter in the forward direction than others direction if the
wavelength is same as particle size.
- A common application of Nephelometry is the measurement of antigen – antibody reaction
because most antigen – antibody complex have a diameter 250 – 1500nm, and wavelength
used 320-650nm .
 Nephelometer:
- A light source
- A collimator
- A monochromatic.
- A sample cuvet.
- A stray light trap.
- A photodetector.
 Turbidimetry:
- This is measurement of the reduction in light transmission caused by formation of particle.
 Application of turbidimetry:
- Used in microbiology analyzer to measure turbidity of sample by detecting bacteria growth
in both cultures :
- Routinely used to measure the antibiotics sensitivity from a such cultures.
- Used in coagulation analyzer to detect clot formation in sample cuvet.
- Used in chemistry to quantify protein concentration in biological fluids such as urine and
cerebrospinal fluid (CSf) .
 Principles:
- Nephelometry and turbidimetry are based on scattering of radiation by particles in the
suspension.
- When a collimated light beam strike a particle in suspension, portion of the light are
absorbed, reflected, scattered and transmitted.