Cerebrospinal fluid (CSF) is a major fluid of the body.

contamination from the external environment = most

likely in tube 1 than tube 2
- to supply nutrients to the nervous tissue,
- remove metabolic wastes, and TUBE 4
- produce a mechanical barrier to cushion the brain
and spinal cord against trauma.  used for cytologic testing
 not done much
meninges - lines brain and spinal cord:
excess from tube 1 to 3 - subject for other test and are
 dura mater (“hard mother”) preserved
 arachnoid mater (“spiderweb-like”)
 pia mater (“spiderweb-like”) STAT - short turn-around time; a more immediate
matter of testing
CSF flows through the subarachnoid space
Mechanical method of specimen preservations are as
blood-brain barrier - keeps csf and blood from mixing follows:
up
TUBE 1: chemistry and serology tubes are frozen
choroid plexus - 20 mL
TUBE 2 at room tep
arachnoid vili - 140 to 170 mL in adults and 10 to 60
mL in neonates TUBE 3 are refrigerated

production of CSF is by: 40% of WBC are lost if unpreserved

 filtration

specimen handling and collection APPEARANCE

lumbar puncture Macroscopic: physical and chemical

neonates - between l4 and l5 Chemical test in CSF is done by automated instruments

aduls - bet. l3 and l4 NORMAL: crystal clear liquid (water-like)

3 sterile tubes specimen Crystal Clear

tube 1 - chemical and serological ttest Cloudy or turbid

tube2 - is for microbiology section Milky

tube 3 - is for cell count because it is the least likely to Xanthochromic

contain cells introduced by the spinal tap collection;
most sterile Hemolyzed or bloody

bloody - intra hernial hemmorhage; traumatic tap -

introduction of the lumpar tap procedure
contamination of blood = tube 1
least likely to contain blood = tube 3
CLOUDY or TURBID – increased protein or lipid
concentration or due to increased protein or lipid
concentration or due to infection with the cloudiness
caused by

TUBE 2
Xanthochromatic – discoloration, presence of different
color other than bloody; pink, orange or yellow
CAUSE : RBC degradation products
Hemolyzed – red or pinkish and clear
= blood is introduced to the sample during collection
If there is only one tube collected = tube is sent first to
microbiology (because that sample has to be free from
contamination prior to microbiology testing)

Bloody – red and clouy, intact blood cells microbio = culture sensitivity, gram stain (secondary)

Yellow – conversion of oxyhemoglobin to unconjugated

bilirubin
TESTS PERFORMED
Red – oxyhemoglobin
1) cell count
Orange – heavy hemolysis METHODOLOGY
- normal adult : 0 to 5 WBCs/L.
Pink – very slight amount of oxyhemoglobin - normal in children and newborn = 30
mononuclear cells/microlites
CAUSE OF XANTHOCHROMIA:  mononuclear = lymphocytes etc
- you can vary the procedure to increase the
Elevated serum bilirubin – autoimmune hemolytic
anemia, transfusion reaction STANDARD NEUBAUER CALC. FORMULA:

*unconjugated bilirubin – not soluble in water, when

increased, are able to cross the blood-brain barrier; if
present it could cause complication in the nervous
system = brain damage
DEPTH OF THE COUNTING CHANBERS: 0.1 MM??
Presence of the pigment carotene = orange
AREA OF CELL CHU CHU = 10 MM^2
Markedly increased protein concentrations and
melanoma pigment 1.1 x 10 = 1mm^3

dilution conversion factor

Xantochromia – immature liver etc *this formula can be used for both diluted and undiluted
specimens.
Refer to the book
FORMULA TO CORRECT FOR THE COLUME
COUNTED:

Traumatic tap

Intracranial Traumatic tap Dilution = performed in bloody sample

hemorrhage collection
Distribution Even (in all 3 Uneven (most Pipette
of blood tubes) bloody tube 1,
least bloody For a clear sample YOU DO NOT DILUTE; less cell in
tube 3) normal cell so it will be lesser if diluted
Color of xanthochromatic Colorless and
supernatant Clear (after
centrifugation)
Clot Negative Positive (after
formation centrifuge)
d-dimer test positive negative
Dilution = Aspirate 30mm of sample to 270 diluent

Not a one step dilution

You cannot lyse RBC with undiluted specimen only
Add it to the 9.9 mL diluted

Serial dilution

1) perform the 1:100 dilution and add 0,m1

mL to the 9.9 mL diluent

1)what is the dilution of a sample prepared by diluting a

50uL-aliquot in 250uL of the diluent

[hysiologic saline solution / normal saline – dilutions for

total cell counts
Corrections for contamination
3% acetic acid to lyse RBC to clearly see WBC
Calculations are possible to correct for WBCs and
protein artificially introduced as a result of a traumatic
tap.
INCC
Ratio for this one = same ratio for wbc
Canal (letter H) = excess fluid will flow CONTRIBUTED BY THE BLOOD TO THE CSF

Add cover slip then add sample or drop the sample Corrected WBC count = actual WBC – WBC(added)

Depth of the cutting chamber = 0.01mm WBC (Added) = WBC (blood) x RBC

1mm by 1mm
DIFFERENTIAL COUNT
PMNS – poly neutrophils
In csf = lymphocytes and neutrophils (more common)
Neutrophils – fine purple granules; 3 to 5 lobes
Granulocytes – neutrophil (abundant), eosinophils,
basophils

Mononuclear – lymphocytes and monocytes = in

neonates

Not a lymphocyte = nucleated RBC

 You do not refrigerated

Darkly staining background

CSF

Turbidimetric methods rely on the precipitation of

protein by either sulfosalicylic acid (SSA) or
trichloroacetic acid.

PROTEIN FRACTIONS

- IgG = increased = blood-brain brarrier defect

- CSF/serum albumin index

CSF GLUCOSE

Normal = 60 – 70% of the plasma concentration (50-

80mg/dL)

Plasma (75mg/dL (57 to 67 mg/dL) CSF glucose

Haemiphillus pneumoniae

Bacterial meningitis

Vira; meningitis – does not decreased CSF glucose only

bacterial and