Cerebrospinal fluid is collected through a lumbar puncture procedure where a needle is inserted into the lower back to draw fluid from around the brain and spinal cord. The fluid is analyzed to diagnose conditions like infections, autoimmune disorders, bleeding, or tumors in the brain or spinal cord. A CSF analysis examines characteristics of the fluid like white blood cell count, proteins, and presence of bacteria or cancer cells to determine the cause of a patient's symptoms. Proper collection and quick transport of CSF specimens to a laboratory is important for accurate diagnostic testing.

1. COLLECTION,TRANSPORT
AND PROCESSING OF
CLINICAL SPECIMENS: CSF

2.  Cerebrospinal fluid is the liquid (colourless)
around your brain & spinal cord.
 The fluid is made by a group of cells, called the
choroid plexus, that are deep inside your brain.
 CSF is a product of plasma filtration and
membrane secretion, and so its composition is
different from that of plasma. CSF, in general, is
clear, colourless, nearly acellular, and has a low
protein concentration.Various ions, enzymes,
and other substances are also found in normal
CSF.

3.  It cushions those organs, picks up needed
supplies from your blood, & gets rid of waste
products from brain cells & provides
lubrication between surrounding bones and the
brain and spinal cord.
 CSF is slightly alkaline and is about 99 percent
water.
 There are about 100 to 150 ml of CSF in the
normal adult human body.

4.  Examination of the CSF may diagnose a number
of diseases.
 A fluid sample is obtained by inserting a needle
into the lumbar region of the lower back below
the termination of the spinal cord; this procedure
is called a lumbar puncture or spinal tap.
 The needle is inserted in the lower back, usually
between the third and fourth lumbar
vertebrae, into the subarachnoid space of
the spinal cord, where the CSF is located.

6.  The kit for collection of CSF should contain:
 Skin disinfectant: 70% alcohol swab and povidone-iodine
 Alcohol with concentrations greater than 70% should not be
used because the increased concentrations result in decreased
bactericidal activity. Do not use alcohol with glycerol added to it.
 Sterile gloves
 Be sure to check the expiration date.
 Sterile gauze
 Surgical mask
 Adhesive bandage
 Lumbar puncture needle
 22 gauge/89 mm for adults
 23 gauge/64 mm for children

7.  Sterile screw-cap tubes
 Syringe and needle
 Transport container
 T-I medium (if CSF cannot be analyzed in a
microbiological laboratory immediately)
 T-I should be refrigerated at 4°C and added to the kit
immediately before use in the field.
 Venting needle (only ifT-I is being used)
 Instructions for lumbar puncture and use ofT-I
medium

8.  Gather all materials from the CSF collection kit and a
puncture-resistant autoclavable container for used needles.
 Wear surgical mask and sterile latex or nitrile gloves that are
impermeable to liquids and change gloves between every
patient.
 Label the collection tubes with appropriate information:
patient’s name, date and time of specimen collection, and
Unique Identification Number. Be sure this number matches
the number on both the request and report forms.
 Ensure that the patient is kept motionless during the lumbar
puncture procedure, either sitting up or lying on the side,
with his or her back arched forward so that the head almost
touches the knees in order to separate the lumbar vertebrae
during the procedure.

9.  Disinfect the skin along a line drawn between the crests of
the two ilia with 70% alcohol and povidone-iodine to clean
the surface and remove debris and oils. Allow to dry
completely.
 Position the spinal needle between the 2 vertebral spines at
the L4-L5 level and introduce into the skin with the bevel of
the needle facing up.
 Accurate placement of the needle is rewarded by a flow of fluid,
which normally is clear and colourless.
 Remove CSF (1 ml minimum, 3-4 ml if possible) and collect
into sterile screw-cap tubes. If 3-4 ml CSF is available, use 3
separate tubes and place approximately 1ml into each tube.
 Withdraw the needle and cover the insertion site with an
adhesive bandage. Discard the needle in a puncture-
resistant, autoclavable discard container.

10.  Remove mask and gloves and discard in an autoclavable container.
 Wash hands with antibacterial soap and water immediately after
removing gloves.
 Transport the CSF to a microbiology laboratory within 1 hour for
culture and analysis.
 If that is not possible, inoculate CSF intoT-I medium (see Section I.C.
below).
 IfT-I is not available, incubate CSF at 35-37°C with ~5% CO2 (see Section
I.D. below) and store in an approved location if the laboratory is closed.
 In the event of a needle-stick injury or other skin puncture or
wound, wash the wound liberally with soap and water. Encourage
bleeding.
 Report a needle-stick injury, any other skin puncture, or any
contamination of the hands or body with CSF to the supervisor and
appropriate health officials immediately as prophylactic treatment
of the personnel performing the procedure may be indicated.

11.  A CSF analysis may include tests to diagnose:
 Infectious diseases of the brain and spinal cord,
including meningitis and encephalitis. CSF tests for
infections look at white blood cells, bacteria, and
other substances in the CSF.
 Autoimmune disorders, such as Guillain-Barré
Syndrome and multiple sclerosis. CSF tests for
these disorders look for high levels of certain
proteins in the cerebrospinal fluid.These tests are
called albumin protein and igG/albumin.
 Bleeding in the brain.
 Brain tumors.

12.  You may need a CSF analysis if you have symptoms of
an infection of the brain or spinal cord, or of an
autoimmune disorder, such as multiple sclerosis (MS).
 Symptoms of a brain or spinal cord infection include:
 Fever
 Severe headache
 Seizures
 Stiff neck
 Nausea and vomiting
 Sensitivity to light
 Double vision
 Changes in behaviour
 Confusion

13.  Symptoms of MS include:
 Blurred or double vision
 Tingling in the arms, legs, or face
 Muscle spasms
 Weak muscles
 Dizziness
 Bladder control problems
 You may also need a CSF analysis if you've had an
injury to your brain or spinal cord, or have been
diagnosed with cancer that has spread to the brain or
spinal cord.

14.  It is done within first half an hour after withdrawal,
because, on longer standing, cells begin to
disintegrate. Normally, 0-10 cells/mm3, all of which
are lymphocytes, which are present. In children, a
normal count is slightly high, about 20 cells /mm3.
 Polymorphs are found only in pathological
conditions.
 Cell count should be done soon after collection of
the fluid.

15.  Centrifuge a little CSF and make 3 smears from
deposit:
 First smear stain by Leishman stain, to observe
leucocytes.
 Second smear with Grams stain and look for
meningococci.
 Third smear with Ziehl-Neelsen stain and look for
acid fast bacilli.

17.  It is done as follows:
CSF Diluting Fluid: Composition
1.Crystal violet - 0.2 gms
2. Glacial acetic acid - 10 ml
3. Distilled water - 90 ml
 Filter if cloudy, add a race of potassium oxalate
 The Fuchs- Rosenthal counting chamber is
specially meant for CSF cell count in entire ruled
area & divide the no. of cells by 3 or multiply by
0.35 to get no. of cells per mm3

19.  Tumor cells, when suspected, should be searched for
after staining the smear of the centrifuged deposit
with pan-cytokeratin stain.
 Count is between 10 to 100 cells/mm3 : In syphilits
disease of CNS, encephalitis, anterior poliomyelitis, in
chronic degenerative diseases of CNS.
 Count is between 100 to 500 cells/mm3: In
lymphocytic choriomeningitis, syphilitic meningitis,
TB meningitis, aseptic meningitis, acute
poliomyelitis, brain tumors.
 Count is over 500 cells/mm3: Count of 1000 or more
with predominance of lymphocytes found in lateTB
meningitis, poliomyelitis, pyogenic meningitis,
influenzal meningitis, brain abscess, etc.

20.  CSF specimens should be transported to a
microbiology laboratory as soon as possible.
 Specimens for culture should not be
refrigerated or exposed to extreme cold,
excessive heat, or sunlight.
 They should be transported at temperatures
between 20°C and 35°C.
 For proper culture results, CSF specimens
must be plated within 1 hour.

22. Thanks for
bearing with
us