Download Blood and Body Fluids

• Blood and Body Fluids
Notes and Labs
2003
Blood
Function: Transport of
dissolved gases
(oxygen and carbon
dioxide), nutrients,
enzymes, etc. In
addition, drugs,
medicines, enzymes
and neurotransmitters
are transported.
Drugs
could include: nicotine,
THC, LSD, cocaine,
methamphetamine,
alcohol. Toxicology
tests for
these.
Neurotransmitt
ers could include:
dopamine, serotonin,
GABA, Ach.
Blood is
composed of cells,
RBC (erythrocytes –
Hb carries oxygen and
carbon dioxide), WBC
(leucocytes from the
immune system– kill
foreign bacteria,
cleaning the blood),
platelets (used in blood
clotting) and
plasma
Blood Types:
A, B, AB, O and Rh
factor
DNA testing
uses the DNA from the
blood cell
nuclei
Hematocrit
measure the % Blood
cells
Spin blood so
that cells go to the
bottom leaving yellow
plasma at the top of the
tube
Plasma (55% of
blood) is a straw
colored part of the
blood which is
composed of water
(90%) and 10%
metabolites, waste,
salts with ions, mostly
Na +, Cl - HCO3- and
proteins.
Tests: Is
it Blood?
In order to
test if a substance is
actually blood, there
are tests that can be
performed using
various chemical
tests.
1. Luminol*:
short uv, spray samples
to see fluorescence
2.
Kastle- Meyer
reagent** + 3 %
H2O2:
wet filter
paper, label with
permanent ink or
pencil
touch stain to
wet filter paper to
transfer the substance
to be tested
add 2
drops of Kastle- Meyer
reagent
Add 2 drops
of fresh 3 % hydrogen
peroxide
Observe
color change will be
pink in the presence of
real blood
3.
Ninhydrin
solution:
Procedur
e:
• Set up samples of white
cloth and white paper
towels with ketchup,
Tabasco sauce,
worceshire sauce, BBQ
sauce,red food dye and
real steak or beef blood
from the
butcher.
*Luminol:
1.0 g luminal + .5 g
Na2CO3 dissolved in
100 ml of distilled
water, then add 0.70 g
sodium perborate (or
0.965 g potassium
iodate can be
substituted).
**
Kastle- Meyer reagent
– This is a reduced
phenolphthalein
solution. KOH,
phenolphthalaein and
Zinc. After dissolved
add ethanol to 500 ml
store in a brown bottle
in the
refrigerator.
Blood
Typing:
• ABO and Rh blood
characterizations are
important in forensic
serology because it can
be done quickly and
relatively
inexpensively. About
80% of the population
are secretors which
means that their blood
type antigens are found
in body fluids other
than blood.
The ABO
Blood System:
Blood
cells can be covered
with polysaccharides
known as antigens
which react with
antibodies found in
blood serum causing
blood cells to
coagulate in an
agglutination reaction
like a chemical
precipitation reaction.
There are 2 basic
antigens A and B. A
person’s RBC can have
either or both or none
i.e. A,B, AB, O. thte
purpose of the
antibodies in the serum
is to attack invaders
that enter the blood
system e.g. viruses.
Antibody coated
viruses are destroyed
by WBC i.e.
vaccines.
Thus, type
A person produces
anti-B antibodies or
anti-B serum to destroy
type B blood cells and
vice versa. Type O has
both anti-A and anti-B
antibodies in its serum
wrong blood type can
be fatal in blood
transfusions and organ
transplants
Generally
41% of a population
are type A, 10% type
B, 4 % type AB and
45% type O.
•
• Testing if agglutinatin
occurs as follows:
• Anti- • Anti• Blood type
A
B
•+
••A
••+
•B
•+
•+
• AB
•••0
• Rh factor is sometimes
referred to as D antigen
Rh+ people have it and
Rh- do not. About 85%
of the general
population has Rh
factor. Potential
problems can occur
between an Rh- mother
and an RH positive
baby. Late in
pregnancy, some of the
baby's blood may cross
over into the mothr's
blood. The mother will
make anti Rh factor
which is not a problem
in the first pregnancy
but could be in
successive
pregnancies. So, the
mother is injected with
anti-Rh antibodies to
eliminate the Rh
antigen before her own
immune system
responds and generates
immunological
memory against the Rh
factor, endangering
any of her future Rh+
babies.
Lab: "Blood
Typing" (used an
ABO Rh kit from
Flinn
Scientific)
Lab:
Blood Spatter
Analysis: Using
simulated blood
(diluted Karo Syrup
with Red and Blue
food dye), students
took platic knifes and
spattered simulated
blood onto white
butcher paper at
different angles outside
the class
room.
BODY
FLUIDS:
• Body fluids can be saliva,
mucous, semen,
vaginal fluid, breast
milk, or urine. These
body fluids can carry
metabolites (waste
products), drugs,
medicines, AB and Rh
antigens, proteins,
hormones, salts
(electrolytes)and
sugars. There exists a
secretory gene (Se) in
about 80 % of the
population independent
of your blood type i.e.
a person could be type
A and Se or type B and
Se etc. This gene
interacts with blood
type genes and
determines your ability
to secrete your blood
type antigens into body
fluids and
secretions. In other
words secretors put
blood type antigens
into the blood and nonsecretors put little to no
blood type antigens
into the blood. Persons
who are secretors can
have their body fluids
tested for blood type.
These people also tend
to have strong immune
systems, therefore it is
better to be a secretor
than a non-secretor.
• URINE: is composed of
95% water and 5% is
other dissolved or
suspended substances
including carry
metabolites (waste
products), drugs,
medicines, AB and Rh
antigens, proteins,
hormones, salts
(electrolytes)and
sugars. In generaly 0.6
to 2.5 liters of urine are
produced by one
person, depending on
intake and environment
i.e. temperature and
humidity. If a person is
diabetic, their urine
may contain glucose.
• LAB tests for urine:
(setup for 3 different
samples)
1.
Tests include, density
(specific gravity),
pH, sugar and
protein.
2.
Teacher Notes:
3.
Students need
approximately 60
minutes to
perform the tests:
4.
Purchase synthetic
urine or make
urine from the
following recipes
1.
A: normal
urine:160
ml of water
+ drops of
yellow food
coloring +
2.0 grams
NaCl
2.
B. add 40 ml of
apple juice
to 66 ml of
normal
urine A
3.
C. Add a few
drops of
egg white
(albumin)
• A crime scenario can be
put in place here
(student or teacher
generated)
• LAB for urine (this lab
based on Lab 3-1
"Don't flush the
evidence", p. 127 of
Crime Scene
Investigations, Walker
and Wood)
1.
Materials:
1.
centrifuge
2.
pyrex test tubes
to hold 20
ml
3.
hot plates
4.
250 ml beakers
(for hot
water baths
)
5.
100 ml beakers
(for urine
samples)
6.
Benedict's
Solution
7.
pH paper
8.
Samples of
synthetic
urine
2.
Procedures:
1.
Obtain crime
scene urine
and urine
2.
3.
4.
from the 2
suspects
Make a Data
Table for
the three
samples and
4 tests and
observation
s: specific
gravity, pH,
albumen (+
or -),
glucose (+
or -), color
odor and
clarity
Record odor,
color
(yellow,
amber,
gold, etc.)
and clarity
(clear,
cloudy,
etc.)
Determine
specific
gravity
(density)
using a
balance and
graduated
cylinder.
Record
results.
5.
6.
Test for pH
using pH
paper and
record
results.
Test for
albumin:
centrifuge
10-15 ml of
each urine
sample
(label each
test tube
with water
proof pen),
remove
supernatant
into 2 clean
test tubes.
Place one in
a hot water
bath for 2-4
minutes,
compare
clarity with
original
sample, if it
becomes
more
cloudy then
albumin is
present and
record a +
in data
table.
7.
Repeat for
other
samples.
Test for
glucose:
Add 10.0
ml of each
sample to a
pyrex
labelled test
tube, add 10
drops of
Bendict's
solution to
each of the
3 samples
and to a test
tub of
water. Place
all four test
tubes into a
hot (not
boiling)
water bath.
Observe
and record
color. If the
blue
changes to
yellow or
gold, then
glucose is
present and
put a + in
your data
table.
• References:
"Biology"
fifth edition Neil A.
Campbell, jane B.
Reece, and Lawrence
g. Mitchell, Addison
wesley Longman.
ISBN 0-8053-65664
intesting site on
urine:
http://www.heartlandh
ealing.com/pages/archi
ve/urine_therapy/
"Cr
ime Scene
Investigations - Real
Life Science Labs for
Grades 6-12" by Pam
Walker and Elaine
Wood ISBN 0-87628135-8 Prentice-hall
(Simon and Schuster)
by The Center for
Applied Research in
education.