+ Dr. Beenish Zaki Senior Instructor Department of Biochemistry Vitamins and its nutritional aspect.
B IOCHEMISTRY P RACTICAL By: Dr. Beenish Zaki. CSF A NALYSIS.
Transcript of B IOCHEMISTRY P RACTICAL By: Dr. Beenish Zaki. CSF A NALYSIS.
BIOCHEMISTRY PRACTICALBy: Dr. Beenish Zaki
CSF ANALYSIS
CEREBROSPINAL FLUID
The surface of the central nervous system is covered by the meninges: Duramater Arachnoid Piamater.
The space between arachnoid and piamater is called sub-arachnoid space.
CSF is secreted by the choroid plexuses. In normal healthy adults rate of CSF formation is
100-250ml/24hrs.
CSF FORMATION AND CIRCULATION
SITE OF WITHDRAWL
•CSF is withdrawn by a procedure called lumbar puncture.•Spinal cord ends near the first lumbar vertebra.•CSF is obtained by passing Lumbar Puncture (L.P) needle between L4-L5.•Second preferred site is L3-L4 into the sub-arachnoid space.
SITE OF LUMBAR PUNCTURE
NORMAL COMPOSITION:
Colour : Clear colorless; no coagulum or deposit
PH : 7.3
Pressure:60-150 mm of Hg
Specific gravity: 1.006-1.002
Cells : 0-4 mononuclear cells/mm3
Protein content: 10-45mg/dl
Globulins: absent
Glucose: 45-100mg/dl
Chlorides: 120-130meq/l
MEASURING OPENING PRESSURE
ABNORMALITIES
In appearance
Blood stained a)Due to traumatic tap RBC under microscope
have normal shape b) In pathological conditions like sub-arachnoid
haemorrhage in this blood is homogeneously mixed with CSF under microscope RBC have a crenated appearence.
ABNORMALITIES CONT’
In appearance Xanthochromia:-Yellow colorization of CSF
due to either: bilirubin Carotenoids
Turbidity :- is due to presence polymorphs/mm3
Coagulum:- In tuberculosis meningitis cob-web like coagulum is seen if the fluid is allowed to stand overnight from the web tubercle bacilli can be examined easily under microscope.
ABNORMALITIES CONT’
In Glucose CSF glucose is increased in :- Diabetes Encephalitis Cerebral abscess CSF glucose is decreased in :- Meningitis due to bacteria. The glucose reduces from the fluid because
the organisms are glycolytic and thus use up the glucose.
A CASE
A 19 year old student presents with fever, headache, nausea, vomiting. On
examination there is neck stiffness.
INDICATION FOR CSF COLLECTION
CSF infection
Hemorrhage
Malignancy
Demyelinating Diseases
CSF PROTEINS ESTIMATION
METHOD AND PRINCIPLE
Method:Turbidity Method
Principle:Proteins are precipitated with Trichloro acetic acid which gives a turbidity.
PROCEDUREProcedure Blank(ml) Test(ml)
Sample - 1.00
ReagentTCA
4.00 3.00
Mix well and take reading immediately using green filter
OBSERVATION AND CALCULATION
O.D of Test= O.D of Standard=0.16 Concentration of sample =
O.D Test X concentration of standard (mg/dl)
O.D Standard
O.D Test x 100
0.16
O.D-Optical Density
REFERENCE RANGE AND CLINICAL CONDITIONS
Adult: 10-45 mg/dl
CSF Protein Increased in: Meningitis Meningoma Acoustic Neuroma Multiple Sclerosis
CSF Protein Decreased in: Malnutrition
CSF CHLORIDE ESTIMATION
METHOD AND PRINCIPLE
Method: Titration Method
Principle:Chlorides present in CSF reacts with Silver nitrate to form silver chloride this reacts with the indicator to form a colored complex
PROCEDURE
Pipette 1ml of CSF and 2ml of distilled water into a conical flask.
Add 3 drops of potassium chromate as the indicator.
Titrate against silver nitrate in the burette to a faint brick red color which is the end point.
OBSERVATION AND CALCULATION
Initial Reading= Final Reading=
Concentration of sample:mEq. Chloride per liter of CSF= ml of silver nitrate solution required x 30
mEq/L= Milliequivalent/L
REFERENCE RANGE AND CLINICAL CONDITIONS
Adult: 90-120 mEq/L
CSF Chloride Increased in: Hypertension Renal disease
CSF Chloride Decreased in: Meningitis
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