Laboratory instrumentation.
Principles of instrumentation:
Impedance .
- Electrical impedance measurement is based on the change in electrical residence
across an aperture when a particles in conductive liquid posses through this
aperture.
Uses of impedance.
To enumerates leukocytes, erythrocytes and platelets in haematology lab.
Typical electrical impedance instrument:
- Aspirate blood is divided into two separate volumes for measurement
- One volume is mixed with diluent and is delivered to the cell bath, where
erythrocytes and platelets counts are performed.
- As a cell posses through the aperture, partially occluding it, the electrical impedance
increase, producing a voltage pulse , the size of which is proportion to the cell size.
- The number of pulse is directly related to the cell count.
Chromatography:
- Chromatography is a separation method based on different interaction of the
specimens compounds with the travel through a support medium .
Techniques.
- Gas chromatography and liquid chromatography.
Principles of chromatography:
- The analyte is combined within a liquid or gaseous mobile phase . usually one phase
is hydrophilic and the other is lipophilic.
- The components of analyte interact differently with these two phase
- This leads to the separation of the different components present in the sample. Each
sample components elutes from the stationary phase at a specific time called as
retention time . retention is the time it take a compound to elute.
- As the components pass through the detector their signal is recorded and plotted in
form of a chromatogram.
Spectrophotometry:
- Is a techniques of measuring the amount of absorbed light through a sample.
Spectrophotometer is an instruments that measure the amount of intensity of light
absorbed by the sample solution as a function of wavelength.
Bouguer developed the principles of absorption spectroscopy in 1700s.
Lambert and Beer:
- Developed the fundamental principles of absorption spectroscopy, commonly
known as Beer’s law.
Beer’s law
- State that, the amount of energy absorbed or transmitted by a solutions is
proportion to the solutions molar absorptive and the concentration of solute.
Principles of spectrophotometry:
- A spectrophotometry is made up of two instruments spectrometry and
photometer.
- The spectrometer is to produce light of any wavelength , while the photometer is to
measure the intensity of light.
The spectrophotometer is designed in a way that the liquid or a sample is place between
spectrometer and photometer.
Photometer measure the amount of light that passes through the sample and delivers a
voltage signal to the display.
- Spectrophotometric method are electromagnetic radiation (EMR), which can take
several forms, the most recognizable being light and radiant heat, others types of
EMR include gamma rays and x-rays, microwaves, radiofrequency, radiation,
ultraviolet radiation.
Components of spectrophotometer.
- Exciter lamps
- Entrance slit
- Monochromator
- Exit slit
- Cuvette
- Photodetector light
- Emitting diode ( LED) display
Function of components:
- The light sources provides the energy that the sample with modify or attenuate by
absorption. The light is polychromatic.
- A wavelength selector or filter isolated a portion of the spectrum emitted by the
sources and focus it on the sample.
- The sample in a suitable container( cuvet) absorbs a fraction of the incident light and
transmit the remainder
- The light that passes through cuvet and sample strikes the cathode of a
photodetector and generates an electrical signal.
