PROPERTIES BLOOD • Determination of the approximate time the

Physical and Chemical crime was committed

Properties and Changes – Blood clotting, Color, Or if blood is already
solidified, etc.

IMPORTANCE OF THE STUDY OF NATURE OF BLOOD

BLOOD 1. Largest circulating tissue of the body
• As circumstantial or corroborative – Fluid like – because it contains vital
evidence substances
• For disputed parentage – especially for 2. Consists of vital substances
those who are looking for custody of – Blood cells, Other nutrients, Electrolytes,
child. Instead of buccal swabs, use blood for – The ones being circulated into our
DNA typing. cardiovascular system
• Determination of the cause of death and 3. Fluid that circulates in to the
the length of time the victim survived Cardiovascular System
the attack – Oxygen, nutrients, etc.
– Pool of blood – the victim is possibly alive
or survived the attack before dying. FUNCTIONS OF BLOOD
– No pool of blood – the victim is dead. 1. Transport of:
• Determination of the direction of escape of a. Gasses (O2 and CO2)
the victim or the assailant b. Nutrients
– Blood spatter or Blood pattern analysis – c. Wastes
how the blood has been spread to 2. Regulates Body Temperature
know where is the origin of the blood or the 3. Regulates pH of the body fluids
position of the victim when the – Metabolic acidosis – can cause disease in
injury was inflicted or how the assailant the body such as for elders is
escaped. arthritis which is too much acidity in the
– Circle – blood drip body.
– Angle - spat 4. Transportation of injected and otherwise
given medicine to the
IMPORTANCE OF THE STUDY OF affected parts of the body
BLOOD
• Determination of the origin of the flow of
blood KINDS OF BLOOD
– Color 1. Arterial Blood:
– Consistency of blood from the crime – Also called as “Capillary Blood”
scene – Bright Red in Color – because it is
– Examples: Bubbly blood – origin is from carrying oxygen.
lungs. – Oxygenated Blood
Brownish – blood is coming from the 2. Venous Blood:
stomach due to addition of acid in the – Contains increased amount of CO2
stomach or the blood is already clotting, the – Dark Red in Color – because it carries
crime is already committed in a few hours or CO2
few days. – Non-oxygenated blood
 B. Liquid Portion (65% of total blood
volume)
CHARACTERISTICS OF BLOOD 1. Plasma: Straw liquid portion of the blood
1. Color – Bright Red (Arterial Blood) 2. Serum: Straw-yellowish liquid when blood
– Dark Red (Venous Blood) is allowed to clot
2. Volume – 70% of Total Body Weight For testing such as drugs, etc.
– 80-85 mL/kg Body Weight
3. Viscosity – Resistance of blood to flow
– Blood is thick and sticky PROBLEMS IN THE STUDY OF BLOOD
– 4-5 times more viscous than water • Where blood has to be searched for.
4. Specific Gravity – Blood = 1.065 due to • Collection, preservation and transportation
cellular elements of specimen suspected to
Water = 1.000 contain blood.
5. pH Reaction – Slightly Alkaline (pH = • Is the stain that of blood?
7.35 – 7.45) • Does the stain contain blood or another
– lower than 7 – acidic substance?
– 7 and above – basic alkaline • Is it human or animal?
• If the stain is of human blood, did of come
COMPOSITION OF BLOOD from the victim, the
A. Formed Elements (35%) Accused or from otherpersons?
1. Red Blood Cells (Erythrocytes)
Number: 5,000,000 RBCs/mm3
Shape: Disc shape biconcave
HEMOGLOBIN (Hb) The Forensics of Blood
– Coloring matter of blood
– Pigment which is found in the cytoplasm
of RBCs FORENSIC ANALYSIS OF BLOOD
- Carries/ transport
1. Visual examination of evidence
2. White Blood Cells (Leukocytes) 2. Presumptive screening test (Is it blood?)
Number: 5,000 – 10,000/mm3 3. Confirmation test (Seriously, is it blood?)
Soldiers of the Body 4. Determine species origin (human blood?)
a. Agranulocytes 5. Identify the blood (whose blood is it?)
– Monocyte
– Lymphocytes PRESUMPTIVE SCREENING TESTS
b. Granulocytes Presumptive Screening Tests – allow a field
– Eosinophil investigator to screen
– Neutrophil evidence to reduce the number of
– Basophil possibilities and to get a
preliminary identification. e.g. Is it blood?
3. Platelets (Thrombocytes) –Negative result means the questioned
Number: 150,000 – 350,000/ mm3 stain is not likely blood
Function: Blood Coagulation – blood clotting –Positive result means the questioned stain
is likely blood
 Some other substance produce colors other
– Presumptive tests produce a color than pink (not blood)
reaction or release of light Still used today
– Tests rely on catalytic properties of
blood (hemoglobin presence) O-Tolidine Test
Positive result = blue
Color Tests
Apply chromogen (color changing chemical) Similar to benzidine; still carcinogenic as it
Apply oxidizing agent (hydrogen peroxide) can be metabolized to
The catalyst of the reaction is hemoglobin benzidine

Rapid color change is a positive result. No longer used: gradually replaced by TMB
This means the stain is blood.
Tetramethylbenzidine (TMB)
Positive result = Blue-green

Most common test for blood

Remember, other non-blood substances 1.Rub stain with moist swab
might catalyze 2. Add TMB
the reaction also. 3. Add peroxidaseLook for quick blue color
– Chemical Oxidants
– Plant materials Leucomalachite Green (LMG) Test

False Positive – A positive result given by Positive result = Green

a substance Not as sensitive as TMB or specific as
that is not blood. phenolphthalein

5 Types of Color Tests Chemiluminescence and Fluorescence

Benzidine
Phenolphthalein –Chemiluminescence – light is emitted as a
O-Tolidine product of the chemical
Tetramethylbenzidine (TMB) reaction
Leucomalachite Green (LMG) –Fluorescence – light is emitted when a
substance is exposed to a shorter
Benzidine Test wavelength of light
–Positive result = blue color
Chemiluminescence and Fluorescence
Carcinogen (cancer causing) in 1974 •More sensitive than color tests
No longer used by sane scientists •May damage blood stain (no blood/DNA
typing)
Tmb •Used to locate and define areas of blood
prob/Kastle – Meyer Test – old blood stains
Positive result = pinkb – cleaned floor
Luminol Teichmann Test and Takayama Test
–Small amount of blood added to
Method similar to color test microscope
slide
Luminol is combined with oxidant and
sprayed over area thought to contain –Chemical solution is added
blood –Slide is heated to form crystals
–Crystals viewed under microscope
Emits a blue-white to yellow green glow
Teichmann Test /Haemin Crystal Test
Fluorescein Takayama Test/Haemochromogen
Crystal Test
Fluorescein is combined with oxidant and
sprayed over area thought to contain blood Determine species origin

Fluoresces is treated with a UV light Most methods test for serum proteins

Fluorescein includes a thickener; this makes Serum proteins are found in all
it animals, but are slightly different
more effective on vertical surfaces
Species ID methods based on
Study showed no interference with DNA antigen/antibody interactions
analysis
Antigen = serum protein

Antibody = produced when foreign serum

protein is detected
CONFIRMATORY TESTS
Certain antibody will only attach to one
Due to the possibility of false positives with species’ serum protein
the presumptive tests, confirmatory
tests are necessary
Antibody is in antiserum
–Actual proof that a stain is blood. Antigen (serum protein) is in
–Establishing the presence of characteristic blood sample
of blood pigment Hb or one of its
derivatives. Human antiserum will only attach to human
–We are actually now testing Hemoglobin blood sample
which is not present in fruit and Rabbit antiserum will only attach to rabbit
vegetables but rather only blood. blood sample
Dog antiserum will only attach to dog blood
Confirmatory tests involve making crystals sample
that detect the presence of hemoglobin
Ring Precipitin
Blood sample (dilute) Blood type is detected
in top layer through an agglutination
test, where antibodies are
Antiserum in heavy added to a blood sample.
bottom layer
- To identify the origin of the species Clumping, or agglutination,
indicates a positive result.
Slide 28
BLOOD TYPING ANALYSIS

Identify the blood (whose blood is it?) Blood sample mixed with 3 antibodies
If blood clumps, proteins are present
GENETIC MARKERS IN BLOOD If no clumps, no proteins are present

If a stain is blood, and it is human blood, ◼ A cells clump with anti-A

then whose is it? ◼ B cells clump with anti-B
–Blood Group Markers ◼ AB cells clump with both
-DNA ◼ O cells do not clump

BLOOD GROUP MARKERS WHY DO BLOOD TYPING?

Many blood group markers including: Less expensive and quicker than DNA
– ABO Markers profiling
– Lewis System Class evidence
– Rhesus System Can be used to link or exclude suspect to
crime scene
BLOOD GROUP (but doesn’t prove guilt)
MARKERS
RH FACTOR
A person’s blood type is
the result of three Rh protein found on RBC’s when Alexander
antigens Weiner worked with
Rhesus monkeys (1940)
The surface of each red Rh+ : 85% Rh- : 15%
blood cell is covered with
proteins called antigens. Rh+ man and Rh- woman = could be
Rh+baby
Type A: has A antigen on surface of RBC First pregnancy : Rh blood of the fetus may
Type B: has B antigen enter the mother’s baby
Type AB: has both A & B antigens The other will develop anti Rh antibodies
Type O: has no antigens on surface The Rh antibodies might burst the blood
cells of the second baby
BLOOD TYPING
SECRETORS
80% of the population are secretors. Their
blood-
type antigens are found in high
concentration in
their body fluids such as saliva, semen,
vaginal
secretions and gastric juice.

GENETIC MARKERS IN BLOOD

◼ ABO blood typing and protein analysis
may help eliminate a
suspect.
◼ Since there are only a small number of
types (ABO = 4 types), a
match does not mean the stain definitely
came from a certain
person
◼ DNA testing can identify a person, and is
becoming just as easy as the above tests