Lab manual
Haemoglobinometry
General Information:
 Module name and code: Blood and body fluids
 Year of the students: The 2nd year
 Time: 1 hours 40 minutes
General instructions and precautions/safety measures:
a. No food and drinks allowed.
b. All students are required to wear lab coats.
c. The students coming ten minutes late in the session will not be
entitled to attendance.
d. Students are not supposed to handle and/or use the scientific
instruments without the permission of the tutor.
Objectives:
At the end of the session, the students should be able to:
a) List the various methods of haemoglobin estimation
b) Describe the principle of Sahli’s method of haemoglobinometry
c) Demonstrate the Sahli’s method of haemoglobinometry
No.
1
2
3
4
5
Materials and equipment’s required for the session *
Sterile lancet
Sterile cotton /gauze swab moist with 70% alcohol or
methylated spirit
Sahli’s haemoglobinometer (haemometer)
Distilled water
Power point presentation
d) Estimate their own haemoglobin level by Sahlio’s method
*All required materials and equipment’s need to be prepared by
technician before the start of session.
Time plan of session
Attendance
Power point presentation
Demonstration of practical
Explanation of the procedure
Asking students to explain the procedure
(learner comprehension)
Asking students to perform the practical by
themselves
Discussion
Total
10 minutes
30 minutes
10minutes
10 minutes
10minutes
20 minutes
10 minutes
100 minutes
Methods:
The practical session will start with comprehensive interactive session
through power point presentation covering all the objectives of the
session. Then the demonstration of the practical by the tutor will take
place followed by other events as mentioned in the time plan. The steps
of the practical demonstration will be as follows:
PROCEDURE:
1. Using the dropper , put drop by drop N/10 HCl in the
haemoglobin tube upto 20% mark .
2. Sterilize the fingertip using spirit swab. Make a bold prick to get
moderately large drop of blood. Suck the blood in the
haemoglobin pipette upto the 20 mm3 mark without any air
bubble.
3. Wipe off any blood sticking to the tip and sides of the pipette
(avoiding touching the bore of the pipette) using cotton.
4. Transfer the blood immediately into the acid taken in the
haemoglobin tube.
5. Rinse the pipette2-3 times with the acid and transfer into the
diluting tube.
6. Mix and keep it undisturbed for 6-8 minutes, so that
haemoglobin gets converted to acid haematin.
7. After 6 -8 minutes, dilute the contents by adding distilled water
drop by drop and mixing the contents after each drop with the
stirrer, till the colour matches with the colour of the standard.
8. Then take the reading both in gram scale and percentage scale by
noting the lower meniscus.
ReferencesTextbook of practical physiology, C.L.GHAI, jaypree brothers Medical Pub; 8thed
(2012)