Instruments used in glucose estimation

 Adjustable micro pipette

 Centrifuge
 Spectrophotometer

1. Adjustable micro pipette:

Adjustable micropipette is an instrument that deliver a measured volume of
liquid from one container to another;

 depending on size, it could be between about 0.1 µl to 1,000 µl (1 ml).

 These pipettes require disposable tips that come in contact with the fluid.

Principle:

The air displacement micropipettes work on the common air displacement principle

Parts of Pipette:

 Push button.
 Push button rod.
 Tip ejector button.
 Gripy/ holder.
 Volumeter.
 Tip ejector collar.
 Tip holder.

Working procedure:

Steps to follow when using a adjustable micropipette

1. Select the volume

2. Set the tip

3. Press and hold the plunger at the first stop

4. Place the tip in the liquid

5. Slowly release the plunger

6. Pause for a second and then move the tip

7. Insert the tip into the delivery vessel

8. Press the plunger to the second stop

9. Pause for 2 seconds

10. Remove the tip

11. Release the plunger

12. Eject the tip into the waste container.

13.
Precautions using Pipette:

 NEVER rotate the volume adjuster beyond the upper or lower range of the
pipette.
 NEVER invert or lay the micro pipettor down with a filled tip; fluid can
run back into the piston.
 NEVER let the plunger snap back after withdrawing or expelling fluid;
this could damage the piston.
 NEVER immerse the barrel of the micro pipettor in fluid.
 NEVER flame the tip of the micro pipettor
 NEVER reuse a tip that has been used to measure a different reagent.
 2. Centrifuge:A centrifuge is a device that uses centrifugal force to separate various
components of a fluid. This is achieved by spinning the fluid at high speed within a
container, thereby separating fluids of different densities (e.g. cream from milk) or
liquids from solids.

Principle:

A centrifuge works by using the principle of sedimentationUnder the influence of gravitational

force (g-force), substances separate according to their density.

Parts of Centrifuge:

 Electric motor.
 A shaft.
 Rotor head on which centrifuge head turns.
 Motor drive assembly.
 Lid.
 Control panel.
 Electric power switch.

OPERATING PROCEDURE
1. Inspect the centrifuge to ensure it is working properly, there is no damage, and that the
centrifuge is able to move freely.
2. Select the appropriate centrifuge tubes or containers, inspect them to make sure there are
no cracks or flaws. Discard any tubes/containers that are cracked or flawed.
3. Fill tubes with the desired liquid.
4. Make sure that the centrifuge tubes are balanced, use a balance to ensure that the weights
of both tubes match.
5. Tightly secure the lids on the centrifuge tubes.
6. Make sure that the outside of the centrifuge tubes are clean and dry before placing them in
the centrifuge.

7. Balance the tubes within the centrifuge.

8. Close the lid. Make sure the lid has a tight fit.
9. Set the run speed and run time. (Never use the rotor’s maximum run speed).
10. Do not leave the centrifuge until it is operating at full speed and the machine seems to be
running smoothly. Ensure that there is no abnormal noise or vibration.
11. If you notice an unusual noise or vigorous shaking, immediately turn the centrifuge off and
remove it from the power source.
12. Once the centrifuge has completed its run, allow the centrifuge to stop its rotation before
opening the lid.

13. After the centrifuge has completely stopped moving, you may remove your samples from
the centrifuge.

Precautions of using Centrifuge:

 Use safety cups whenever possible

 Disinfect or clean weekly and after all spills or breakages.

 Lubricate O-rings and rotor threads weekly.

 Do not operate the centrifuge without the rotor properly

balanced.

 Do not use rotors that have been dropped.

 3. Spectrophotometer:

The spectrophotometer is an instrument that measures the amount of intensity

of light absorbed by the sample solution as a function of wavelength. This technique of
measuring the amount of absorbed light through sample solution is known as
spectrophotometry.

Principle: Spectrophotometer is based on the Beer-Lambert Law which states that the amount
of light absorbed is directly proportional to the concentration of the solute in the solution and
thickness of the solution under analysis. 

A = ∈cl

Where A = absorbance, c = concentration, l = path length 

Where ∈ = absorption coefficient 

Parts of Spectrophotometer:

 Light source.
 Prism/ monochromator.
 Slit/filter
 Cuvette/ sample holder
 Detector
Working Procedure of Spectrophotometer: