AUBF CSF

Cotugno

Cerebrospinal Fluid was first recognized by _____ in 1764

1764

Cerebrospinal Fluid was first recognized by Cotugno in _____

2 lumbar ventricles, 3rd ventricle, 4th ventricle

CSF Produced by Choroid Plexuses of the

blood capillaries in the Arachnoid Villi/Granulation

CSF is reabsorbed in

90 - 150 mL

TOTAL VOLUME of CSF:

• Adults:

10 – 60 mL

TOTAL VOLUME of CSF:

Neonates:

20 mL

Rate of Production of CSF:

20 mL

Rate of Absorption of CSF:

Blood-Brain Barrier

tight-fitting junctures of endothelial cells in choroid plexuses that prevent passage of many molecules

Chemistry

Tube 1

Microbiology

Tube 2

Hematology

Tube 3

Additional Tests

Tube 4

microbiology

If only one tube can be collected, it must be tested first by _______

1 and 4

It is not unusual for cell counts requested to be performed on both Tubes _____ to check for cellular contamination by the puncture.

Colorless

Color

Same with water

Viscosity

Crystal clear

Clarity

1.006 to 1.008

Specific Gravity

7.30 to 7.35

pH

50 to 200 mm H20

Pressure

Slightly Hazy

200 to 500 wbc/uL

Cloudy

Above 500 wbc/uL

Turbid/Milky

wbc, rbc, proteins, increased lipid concentration, microorganisms, aspirated epidural fats, radiographic contrast media

Clotted

Froin’s Disease (increased proteins and clotting factors)

traumatic tap

Non-Pathologic cause of bloody appearance of CSF

intracranial hemorrhage or subarachnoid hemorrhage

Pathologic cause of bloody appearance of CSF

Viscous

Cryptococcal Meningitis metastasizing mucin producing adenocarcinoma

Cryptococcal Meningitis

________ metastasizing mucin producing adenocarcinoma

Oily

Radiographic Contrast Media

Pellicle Formation

Tubercular Meningitis/ web-like upon refrigeration

Xanthochroma

Presence of RBC Degradation Product

Pink

Color of CSF with very slight amount of oxyhemoglobin

Orange

Color of CSF with heavy hemolysis

Yellow

Color of CSF when there is conversion of oxyhemoglobin to unconjugated bilirubin

Brown

Methemoglobin, Hematoma, Melanin

Hazy/Turbid/Cloudy/Milky

Increased WBCs (>200/uL)
Increased RBCs (>400/uL)
Lipids & protein microorganisms

Bloody

Increased RBCs (>6,000/uL)
Traumatic Tap
Intracranial hemorrhage

Increased carotene,
Increased melanin,
Increased protein (>150 mg/dL),
Rifampin

other causes of xanthochromia

3% Glacial Acetic Acid

Specimens requiring dilution use

0 -5 wbc/uL

Normal CSF WBC Count:

• Adult:

0-30 mononuclears/uL

Normal CSF WBC Count:

• Neonates:

1:10

Dilution

Slightly Hazy

1:20

Dilution

Hazy

1:100

Dilution

Slightly Cloudy

1:200

Dilution

Slightly Bloody or Cloudy

1:10,000

Dilution

Bloody or Turbid

• 1st: Concentrated
• 2nd: Smeared
• 3rd: Stained

When performing differential count, the specimen should be:

Cytocentrifugation

fluid is added to a conical chamber

cells are forced into a monolayer within a 6mm diameter circle on the slide

0.1 mL of CSF + 1 drop of 30% Albumin

produces an adequate cell yield when processed with cytocentrifuge

0.2 mL of CSF + 2 drops of 30% albumin

daily control slide for bacteria

Lymphocytes
Monocytes
Neutrophils

NORMAL CELLS in CSF:

• 70:30

L: M Ratio in ADULTS:

• 30:70

L:M Ratio in Infants:

80

Up to __% monocytes is considered normal

PLEOCYTOSIS

increased number of normal cells an abnormal state

• immature WBC
• eosinophils
• plasma cells
• macrophages
• increased tissue cells
• malignant cells

ABNORMAL CELLS in CSF:

15 – 45 mg/dL

Normal Values of Protein

Adults

150 mg/dL

Normal Values of Protein

500 mg/dL

Normal Values of Protein

Albumin

– major CSF protein

Pre-albumin

2nd Most prevalent (1st only in migration in electrophoresis)

Alpha-globulins

Haptoglobin & Ceruloplasmin

Beta-globulins

Tau Protein (AKA: Beta2-Transferrin)
Carbohydrate-deficient transferrin

Beta2-Transferrin

Tau Protein is also known as

Gamma-globulins

IgG and some IgA

IgM, Fibrinogen, Beta Lipoproteins

Not found in normal CSF

1. Dye-binding
2. Turbidity production

Methods for Total Protein Determination

Turbidity Method

adapted in automated instrumentation in the form of nephelometry

Trichloroacetic Acid (3% TCA)

1. Turbidimetric:

• preferred method
• precipitates BOTH albumin & globulin

Sulfosalicylic Acid (3% SSA)

1. Turbidimetric:

• precipitates albumin ONLY

sodium sulfate

In Sulfosalicylic Acid (3% SSA) to precipitate globulin add

Dye-binding

• protein binds to dye (red to blue)
• Result: Increased protein = Increased blue color

Coomasie Brilliant Blue

dye used in dye binding

CSF/SERUM ALBUMIN INDEX

Evaluates BBB integrity

• Normal Index value of INTACT BBB

9-14

Index Value BBB
slight impairment

15-30

Index Value BBB
moderate impairment

> 30

Index Value BBB
severe impairment

100

Index Value BBB
complete damage to BBB

CSF IgG INDEX

• Measures IgG synthesis within the CNS

Normal CSF IgG INDEX

>0.77

CSF IgG INDEX IgG production within CNS

Electrophoretic Techniques

detection of oligoclonal bands

This is the METHOD OF CHOICE when it is necessary to determine if the fluid is CSF.

Agarose gel electrophoresis followed by CBB staining

Most frequently performed Electrophoretic Techniques

MULTIPLE SCLEROSIS

a demyelinating disorder

tau (B2 – Transferrin)

This is the METHOD OF CHOICE when it is necessary to determine if the fluid is CSF.

How? Through __________ identification

• (+) Anti-myelin Sheath Autoantibody
• (+) Oligoclonal band in CSF but not in serum
• (+) Myelin Basic Protein
• Increased IgG Index

Finding in Multiple Sclerosis

Myelin Basic Protein (MBP)

protein component of the lipid-protein complex that insulate the nerve fibers

Used to monitor the course of multiple sclerosis

myelin sheath

Presence of MBP in CSF indicates destruction of ______

60 – 70 % of blood glucose (50 – 80 mg/dL)

Glucose

Normal Values

VIRAL MENINGITIS

Glucose is Normal in:

blood glucose

CSF glucose is done in conjunction with

2 hours prior

Specimen for blood glucose should be drawn _____ to spinal tap to allow EQUILIBRATION between CSF and plasma glucose

• 10 - 22 mg/dL

Lactate Normal value

BACTERIAL MENINGITIS (>35 mg/dL)

TUBERCULAR & FUNGAL MENINGITIS (>25 mg/dL)

Hypoxia

Lactate is elevated in

GLUCOSE

Lactate is inversely proportional to

Lactate

Sensitive method for evaluating the effectiveness of antibiotic therapy

Glutamine

Product of Ammonia and a-ketoglutarate metabolism

Indirect test for the presence of excess ammonia in the CSF

Ammonia and a-ketoglutarate

Glutamine is the product of ______ and ____ metabolism