COLLEGE OF NURSING

DEPARTMENT OF MEDICAL TECHNOLOGY

OPERATING A MANUAL SPECTROPHOTOMETER MEDT 25 : INTERNSHIP

SKILLS ENHANCEMENT

I. INTRODUCTION

The spectrophotometer is an instrument which measures the amount of light that a

sample absorbs. A spectrophotometer is made up of two instruments: a spectrometer and a
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 placed between spectrometer and photometer. The photometer measures the
amount of light that passes through the sample and delivers a voltage signal to the display.

The basic spectrophotometer instrument consists of a light source, a digital display, a

monochromator, a wavelength sector to transmit a selected wavelength, a collimator for
straight light beam transmission, photoelectric detector and a cuvette to place a sample.

II. PRINCIPLE

The spectrophotometer works by passing a light beam through a sample to measure the
light intensity of a sample. These instruments are used in the process of measuring color and
used for monitoring color accuracy throughout production.

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 COLLEGE OF NURSING
DEPARTMENT OF MEDICAL TECHNOLOGY

III. IMPORTANCE

Spectrophotometer is a highly versatile tool, which is used to determine the concentration

of solid particles in any suspension, especially a blood sample, or transmittance of a
solution. It can also be routinely utilized for data analysis as well as research purposes.

IV. MATERIALS

• Spectrophotometer Incubator
• Pipette Cuvettes
• Pipette tips Reagents with standard
• Timer Sample from venipuncture
• Distilled water

V. PROCEDURE

1. Turn on the power switch. Warm up the machine for 15 minutes.

2. Set the wavelength for the desired test (see insert).
3. Prepare the cuvettes for incubation. Make sure that all cuvettes are clean and free from
dirt/ finger prints. Do not hold the cuvette in its clear sides.
4. Proper order for cuvettes with their corresponding contents are as follows:
(1- water blank) (2- reagent blank) (3- standard) (4- sample)

*Refer to the insert for the amount/ volume required.

5. Set the timer for incubation (see insert for incubation time).
6. Place each cuvettes in the cell rack/ holder found in the sample room of the machine
according to their proper order then close the window cover.
*Ensure that transparent sides of the cuvette are in the optical path.
7. Gently pull the rod (located just below the sample room) and stop at the first click. The
value should be at zero.
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COLLEGE OF NURSING
DEPARTMENT OF MEDICAL TECHNOLOGY

8. Once again, pull the rod then stop at the click. Value should still be zero.
9. Repeat the same procedure (no.8). This time, record the value.
10. Pull the rod for the last time and stop at the click. Record the result for your sample.
11. Compute for the standard and sample value to get the actual result as indicated in the
reagent insert.
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