Scribd
Upload
91 views48 pages

Introduction To Biochemstry Lab

The document provides instructions for laboratory safety and procedures. Key points include wearing personal protective equipment like lab coats and closed-toe shoes. Proper labeling and disposal of materials is required. Safety equipment like fume hoods and eyewash stations must be available. Procedures for blood collection, use of pipettes and centrifuges, and distillation are outlined. Common laboratory instruments such as spectrophotometers are also described.

Uploaded by

Nas Kata
Copyright
© Attribution Non-Commercial (BY-NC)
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPT, PDF, TXT or read online on Scribd
91 views48 pages

Introduction To Biochemstry Lab

The document provides instructions for laboratory safety and procedures. Key points include wearing personal protective equipment like lab coats and closed-toe shoes. Proper labeling and disposal of materials is required. Safety equipment like fume hoods and eyewash stations must be available. Procedures for blood collection, use of pipettes and centrifuges, and distillation are outlined. Common laboratory instruments such as spectrophotometers are also described.

Uploaded by

Nas Kata
Copyright
© Attribution Non-Commercial (BY-NC)
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPT, PDF, TXT or read online on Scribd
You are on page 1/ 48

Introduction to biochemistry

lab
LABORATORY RULES AND
INSTRUCTIONS
1. Prepare for the experiments by reading the
laboratory manual before coming to the lab.
2. Laboratory coats, safety glasses and closed in
shoes must be worn at all times. Gloves are
provided and must be worn when handling
biological materials.
3. Eating, drinking and smoking in the laboratory
are strictly prohibited.
4. Do not waste reagents and supplies. Some are
expensive and may be required for following
groups.
5.Return all apparatus, reagents and samples to their proper place, so
that others may use them.
6. Label all tubes etc. with a permanent marker pen for easy
recognition. Used test tubes should be placed in the buckets
provided and used glassware left at the ends of the benches.
7. Dispose of all pipette tips into the containers provided on the
bench.
8. Sharps (broken test tubes, needles, scalpel blades etc.) must be
placed in the yellow sharps containers.
9. Ensure you fully understand the correct use of all
instruments/apparatus before you start to use it. Use all apparatus
carefully.
10. Rinse all cuvettes that you have used and place them into the
special wash bucket
11. Turn off all electrical equipment at the end of the laboratory
session.
12. Make sure your bench is tidy and has been washed .
13. WASH YOUR HANDS BEFORE LEAVING THE LABORATORY.
Safety Equipment
• The following safety equipment is most
often used in laboratory settings:
– Eye protection
– Lab coats
– Gloves
– Fire protection equipment
– Materials storage cabinets
– Eye wash/shower
– Fume hood
Eye Protection
• Safety glasses, safety
goggles, or face shield
• Used to protect eyes
and face from spills or
splashes (one of the
most common
accidents in labs)
Eye Protection
• Face shields offer
the most protection
• Scratches can be
avoided by never
laying the protection
face down
Lab Coats
• Protect body and
clothes from spills
• Contribute to a sterile
environment
Gloves
• Usually surgical style, thin plastic
• Offer some protection to skin
• Used to achieve aseptic conditions
Materials Storage Cabinets
• Flammables
– Isolates flammable
chemicals for safety
– Should contain
chemicals ONLY
– Fire resistant
Eye Wash/Shower

• Should only be used in


case of emergency
– After chemical exposure,
eyes should be rinsed for
more than a minute to
ensure that damage is
limited as much as
possible.
Fume Hood
• Removes fumes
produced by
chemical solutions
from the laboratory
Chemical Safety
• Make certain that
chemicals are clearly
labeled with Material
Safety Data Sheets
(MSDS) well marked
and easily accessible.
Chemical Spills and Exposure
• Spills should be quickly
contained and the area
secured
• Special media can be
used to absorb harmful
chemicals
• Any exposed skin should be immediately
rinsed or neutralized (in the case of acid or
base)
• particularly with eye exposure MSDS
should be consulted for treatment
– A mild acid can sometimes be used to
neutralize areas exposed to a strong base
(works both ways)
Blood collection:
• Blood specimen collection is performed routinely to
obtain blood for laboratory testing.
• Blood can be obtained from venous access
devices(venipuncture). and sometimes by fingerstick.
• Before puncturing, the patient's skin should be cleaned.
iodine can be used, or alcohol.
• There are three veins in the antecubital area that are
appropriate for venipuncture: the medial,cephalic,and
basilic veins.
• The vein of choice is the medial, because it is usually the
closest to the skin's surface, the largest, and usually the
least painful to puncture.
• If the antecubital sites cannot be accessed, wrist and
hand veins are also acceptable for venipuncture
• Normally, a 21-gauge needle is used to collect blood.
• Never insert the needle at greater than a 30 degree
angle.
• Avoid drawing blood from an arm affected by a stroke or
neurological injury that has resulted in a loss of
sensation.
• collect the blood specimen from the opposite arm if a
patient is receiving intravenous fluids.
• Do not use a site that is swollen.
• Remove the tourniquet when the final tube of blood to be
drawn is filling
Tying the Tourniquet
1. 2.

3. 4.
If no flow - common causes

Bevel near or in
vein wall
Hematoma formed

Needle passed
through vein
Vein collapsed
Send the specimen to the
laboratory as soon as possible.
• Blood collection tubes come with a variety
of colored stopper caps, and may contain
additives.
• The following tubes are the most
commonly used types:

• Red top. This tube contains no additives.


It is used for a variety of tests, including
blood typing and crossmatching.
• Serum separator tube (SST). This tube
contains a polymer gel and clot activator. The
SST is commonly used for blood chemistries.

• Lavender top. These tubes are used primarily


for obtaining complete blood counts. They
contain EDTA, an anticoagulant additive that
chelates calcium.
• Dark green top. Green top tubes contain the
anticoagulant heparin and are often used to
obtain lithium and ammonia levels.

• Light blue top. The light blue top tubes contain


sodium citrate, an agent that removes calcium,
and are used to obtain (PT) and (PTT).

• Light gray top. This specimen tube contains


sodium fluoride and potassium oxalate,
antiglycolytic agents that preserve glucose for up
to five days. The tube is used primarily to obtain
glucose levels.
Order of the Draw
• Red
• Blue
• SST
• Green
• Lavender
• gray
• Patient name and medical record
number
• Specimen requirements
• Collection date and time
• Volume
• Ordered Test
• Department codes
Type of sample
1. Whole blood : plasma + RBC’s
2. Plasma: serum + coagulation factor
3. Serum: no coagulation factor
• hemolysis:- The destruction of red blood
cells, which leads to the release of
hemoglobin from within the red blood cells
into the blood plasma.
• Lipemic sample: high cotent of lipid wich
interfer with many test measurment.
fingerstick

• Venipuncture is not always possible or


appropriate for the blood specimen required.
• In these cases, a fingerstick or heelstick may be
the method of choice for obtaining the specimen.
• The best locations for fingersticks are the third
and fourth fingers of the patient's non-dominant
hand
Finger Sticks and Capillary Draws
• Avoid sticking areas that are thick, where
there is little soft tissue, or where the bone
is close to the surface. Do not puncture a
finger that is swollen.
• The first drop of blood should be wiped
away, as this first drop tends to contain
extra fluid from the tissues.

• Instead of firmly milking the finger, it


should only be gently massaged to gain
drops of blood.
Laboratory Equipment
• There are several different types of
equipment that are essential to labs:
– Autoclave
– Centrifuge
– Micropipette
– Compound Light Microscope
– Incubator
– Water Bath
– Hot Plate
– Shaker
– Graduated Cylinder
Autoclave
• A chamber that uses high levels of heat
and pressure to sterilize instruments and
materials, or destroy harmful
organisms/pathogens.
Specimen separation :Centrifugation

• A packing device for separating components within a liquid or from a


liquid

• Separation activity is a function of both centrifugal force and timing

• Proper balance, lubrication and rotor function are essential for proper
centrifugation to occur
• close centrifuge firmly.
• select velocity and time

• Never open the centrifuge while the rotor is moving or try to stop the
rotor with your hands
Micropipette
• An instrument used to measure
and extract very small amounts
of liquid from a solution.
– Different versions measure to
different levels of accuracy, but
usually to the nearest uL
(microliter)
Using a Micropipette
• To avoid air bubbles
and extract the correct
amount of solution
utilizing a micropipette,
the tip must be
completely submerged
in the solution.
Water Bath
• A vessel that uses water
to heat or maintain a
constant temperature of
laboratory materials or
equipment.
• Often used for
incubation
Hot Plate
• Instrument that applies direct
heat to glassware containing
laboratory solutions
Graduated Cylinder
• Used to measure the
volume of liquids
• Readings should always
be taken at the
MENISCUS, the lowest
part of the curve.
distillation
D.H2O has been purified to remove
almost all organic materials, by using
distiller in which water is boiled and
vaporized.
• Many impurities do not rise in the water
vapor, remaining in the boiling apparatus.
• - Problem with distillation for preparing
reagent water includes the carryover of
volatile impurities and entrapped water
droplet that may contain impurities into the
purified water.
spectrophotometer

• A spectrophotometer is an instrument for


measuring the absorbance of a solution
• Light source: tungsten lamb
• Monochromators: isolation of individual
wavelengths
• Sample cell: fixed size
• Photo detector: convert light electrical
energy
Beer's law
• State that the concentration of substances
is directly proportional to the amount of
light absorbed or inversely proportional to
the logarithm of the transmitted light.
• A=ЄXbXc
• Є = molar absorpativity
• b = length of light
• c = concentration of absorbing
molecules.
• Standard substance: purified material of
known concentration.
• Control material: A material to be used for
the assessment of the performance of an
analytical procedure of known rang of
concetration , not high purified.

You might also like