Name: ____________________________ Date Performed: ______________

Section /Group No.: ________/_______

Score_______________

Experiment No. ____

CONFIRMATORY TESTS FOR BLOOD
I. Objective: To establish the presence of the blood pigment or hemoglobin.
II. Materials: Microscope, fresh blood, blooadstained material, glass slide, glass cover
III. Discussion:
The presence or absence of blood stains often provides important information for those
investigating criminal cases. For this reason, forensic scientists are often called on to determine whether or not
a particular stain is blood, and if so, whose. The detection of blood is usually based on one of three classes of
methods.
Crystal tests-Haem forms crystals when reacted with certain reagents. The most common such reagent is
pyridine, which forms characteristic pink crystals.
Catalytic tests-These tests rely on the fact that haem can catalyse the breakdown of hydrogen peroxide.
As the H2O2 breaks down, another substance in the reaction mixture is oxidised, producinga colour change. It
is important to note that a positive test does not mean that a given stain is blood, let alone that it is human
blood, as various enzymes and certain metals can also give positive results.
Instrumental methods- Chromatography can be used to identify the presence of haemoglobin. These tests
are used practically for several different purposes. These include both the confirmation of the nature of visible
stains (i.e. that they probably are or definitely are not blood), the detection of non-visible stains (e.g. on plants
or washed clothing) and the enhancement of hard to see stains. Stain enhancement is useful for situations
where a footprint, handprint, fingerprint etc. is faintly outlined in blood, as chemical methods can enhance
that stain so that the print can be measured and matched with suspects. In all of these tests it is important to
ensure that the chemical reactions do not prevent later tests being done to help to identify who the blood
belongs to.

IV. Microscopic Examination

A. 1. Place a drop of human blood on a glass slide and cover with glass slip.
2. Observe under the microscope.
3. Draw and identify by labeling the parts of human blood,
erythrocytes,leucocytes,thrombocytes and
plasma.Repeat the procedure for animal blood.

Human blood Animal blood

4. Describe th red blood cells of human

blood.__________________________________________________
___________________________________________________________________________________
 ___________________________________________________________________________________

5. Describe the red blood cells of animal

blood._________________________________________________
_____________________________________________________________________________________

B. Microchemical/Microcystalline Test
TAKAYAMA TEST or HAEMOCHROMOGEN TEST
( Chemical Reagents;Takayama reagent( 3ml of 10% NaoH,3mL pyridine,3mL saturated
glucose soln. and diluted
w/7mL distilled water); 0.9% saline solution
1. Cut a piece of stained material and place it on a glass slide.
2. Add 3 drops of saline solution and tease the stain with needle. Add 3 mL more drops of
saline solution and allow
to stand for 5 minutes.
3. With the use of needle, spread over the glass slide so as to have a very thin layer.
4. Add a drop of Takayama reagent on the stain and cover with glass slip. Heat gently over
on an alcohol flame to
speed up the reaction.
5. Observe the treated specimen under the microscope.
Observation as seen under the microscope.

5.1 Describe the shape of the crystals formed.__________________

_____________________________________________________
_____________________________________________________
5.2 What is the color of the crystals?___________________________
5.3 What do you call these characteristic crystals formed when blood
Reacted with the reagent?
___________________________________________________________________
___________________________________________________________________________________
TEICHMANN TEST
( Chemical Reagents- NaCl, Glacial acetic acid )
1. Place a minute fragment of the stain on a glass slide. Add a small crystal of sodium
chloride and 2 to 3 drops
of acetic acid. Place cover slip and heat gently over small flame ro evaporate the acid.
Cool, then examine
under high power objective lens.
A. Describe the shape of the crystal
formed.______________________________________________________
___________________________________________________________________________________
B. What is the color of the crystals?
__________________________________________________________
Guide Questions:
1. What is the the importance of microscopic examination test from criminalistic stand point?
______________________________________________________________________________________
______________________________________________________________________________________
2. What substance in the red blood cells that is responsible for the red color of blood?
______________________________________________________________________________________
3. In microcrystalline test, what substance confirms that the specimen in really blood?
 _________________________________________________________________________________________
_______________________________________________________________________________________
__________________________________________________________________________________________
4. Aside from Takayama and Teichmann test, what other microchemical/microcrystalline test
can be applied to
identify blood?
_______________________________________________________________________________
_______________________________________________________________________________________
_______________________________________________________________________________________
CONCLUSION:

Name: ____________________________ Date Performed: ______________

Section and Group No.: ________/_______
Score_______________

Experiment No. ____

BLOOD GROUPING OF FRESH BLOOD

I. Objective: To study the technique of grouping fresh blood by direct method.

II. Materials and Reagents: lancet, sterilized pin, 0.9% saline solution, Anti-A and Anti-B sera,
glass slides with well.
III.Procedure:
1. Using sterilized pin,needle or lancet, prick the finger or ear lobe of the subject whose blood
group is to be
determined.
2. Put 0.5mL of th fresh blood specimen into a clean test tube.
3. Put 0.5ml saline solution and then shake.
4. Centrifuge for 3 minutes.
5.Remove the supernatant liquid and add for the second time same volume of saline solution.
6. Centrifuge for the second time if the supernatant liquid is turbid and repeat step 5.
7. Suspended red blood cells in saline solution. Be sure it is bright red color.
8.Get 2 glass slides and place a drop of anti-A and anti-B on each separate slide.
9. To each slide well, add a drop of the unknown fresh blood specimen.Place in a shaker for two
minutes to three
minutes and observe under the microscope.
IV. Observation:
1. Draw what has observed under the microscope.
 A B

2. What happened when the respective serum was added to the red blood cells?
_______________________________________________________________________________________
_______________________________________________________________________________________
_______________________________________________________________________________________

3.What is agglutination?
___________________________________________________________________
_______________________________________________________________________________________
_______________________________________________________________________________________

4. Why does agglutination takes place?

_______________________________________________________
_______________________________________________________________________________________
_______________________________________________________________________________________
5. Why is the unknown fresh blood specimen washed with saline solution?
_______________________________________________________________________________________
________________________________________________________________________________________
________________________________________________________________________________________
6. Why are the red blood cells suspended in saline solution when grouping fresh blood?
________________________________________________________________________________________
________________________________________________________________________________________
________________________________________________________________________________________
________________________________________________________________________________________
7. What is the group of the unkown fresh blood specimen?
________________________________________________________________________________________
________________________________________________________________________________________
Fill in the table using plus (+) or minus (-) signs to prove that your answer in number 7
is correct.

Blood Group Anti-A Anti-B

A
B
AB
O

CONCLUSION: