DR.RITTU CHANDEL
MD BIOCHEMISTRY (SECOND YR)
GRANT GOVT MEDICAL COLLEGE, BYCULLA, MUMBAI –
400008
29 – 10 -2013
Cerebrospinal fluid

Total volumes:
– Adults: 140 - 170 ml
– Children:
29-Oct-13

cerebral ventricles – 30ml
subarachnoid space - 120 ml

10 - 60 ml
Dr.Rittu Chandel

2
hydrocephalus

29-Oct-13

Dr.Rittu Chandel

3
Functions of CSF
Buoyancy
2. Protective effect
3. Metabolic
4. homeostasis
1.

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Dr.Rittu Chandel

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Specimen collection

Cisternal (occipital)
In paralysis
Ventricular puncture
(lateral occipital)
For infants
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Dr.Rittu Chandel

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CSF analysis
CSF

Gross

microbiology
29-Oct-13

pathology

Dr.Rittu Chandel

biochemistry
6
Polypropylene tubes

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Dr.Rittu Chandel

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composition
•
•
•
•
•
•
•
•

Colour
- Colourless
PH
- 7.28 – 7.32
Appearance
- Clear
Sp. Gravity
- 1.003 – 1.004
No clot formation on standing
Proteins
- 10 – 45 mg/dl
Glucose
- 45 – 100 mg/dl
Chlorides
- 120 -130 mEq/l

29-Oct-13

Dr.Rittu Chandel

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







Urea
Uric acid
Creatinine
Cholesterol
Ammonia
A:G ratio
Serum:CSF protein

29-Oct-13

Dr.Rittu Chandel

- 6.0 - 16 mg/dL
- 0.5 - 3.0 mg/dL
- 0.6 - 1.2 mg/dL
- 0.2 - 0.6 mg/dL
- 10 – 35 μg/dL
- 3:1
– 200:1
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Gross examination
 Appearance

Normal – crystal clear, colourless and no coagulum
1.Colour
Any change in colour is always pathological
Red
Yellow

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Dr.Rittu Chandel

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xanthochromia
 Pale pink to yellow colour in supernatant of CSF

Froin’s syndrome
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Dr.Rittu Chandel

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D/D of bloody CSF
 Traumatic tap - hemorrhagic fluid clears between

first and third tubes; remains uniform in SAH
 Pathological
- RBC have crenated appearance
 Xanthochromia, erythrophagocytosis and hemosedrin

laden macrophages indicate a subarachnoid bleed in
absence of prior trauma

29-Oct-13

Dr.Rittu Chandel

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Gross examination(contd)
 2. Turbidity
o WBC >200 cell/μl
o RBC >400 cell/μl
o Microorganisms, radiographic contrasts, aspirated

epidural fat
o Protein > 150 mg/dl
 Clot formation
 Viscous CSF

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Dr.Rittu Chandel

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Pressure of CSF
Normal
Adults – 90 – 180 mm water
Infants and children – 10 – 100 mm water

>250 mm water
Intracranial hypertension
Tumors

Decreased pressure
Dehydration
Circulatory collapse
CSF leakage
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Dr.Rittu Chandel

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Biochemical analysis of
CSF
 1. Glucose

Normal – 45 – 100 mg/100 ml
decreased – coccal meningitis
TB meningitis
intrathecal streptomycin
Increased – diabetic hyperglycemia

In assessing response to treatment
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Dr.Rittu Chandel

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 2.Proteins

Normal – 15 – 45 mg/dl
Term infants – 90 – 150 mg/dl
Pre term - 115 – 170 mg/dl
 Increased CSF protein
I. Increased permeability of blood brain barrier
II. Increased intrathecal IgG secretion

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Dr.Rittu Chandel

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electrophoresis

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Permeability of blood brain
barrier
CSF/serum albumin index
index

permeability

<9

intact barrier

9 – 14

slight impairment

14 – 30

moderate impairment

>30

severe impairment

Index slightly elevated in infants up to 6 months of age
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Dr.Rittu Chandel

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CSF serum immunoglobulin
ratio

 Increases in multiple sclerosis

 Increased CSF IgM and kappa light chains – marker of

multiple sclerosis
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Dr.Rittu Chandel

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CSF proteins and CNS
diseases
proteins

Diseases or disorders

Α2 - macroglobulin

Subdural hemorrhage, bacterial meningitis

Β – amyloid protein 42 and ζ protein

Alzheimer’s disease

Β 2 - microglobulin

Leukemia/lymphoma, bechets syndrome

C- reactive protein

Bacterial and viral meningitis

fibronectin

Lymphoblastic leukemia, AIDS, meningitis

methemoglobin

Mild subarachnoid/subdural hemorrhage

Mylein basic protein

Multiple sclerosis, tumors

Protein 14- 3 -3
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Dr.Rittu Chandel

Creutzfeldt – jacob disease

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CSF leak (rhinorrhea)

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Dr.Rittu Chandel

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CSF lactate
 CSF and blood lactate are largely independent
 Normal

newborn – 10 – 60 mg/dl
Child and adult – 9 – 26 mg/dl
Differentiating meningitis

CSF lactate
(mg/dl)

meningitis

< 25 mg/dl
Almost always less
than 35 mg/dl

viral

>35 mg/dl

bacterial

Persistently elevated - poor prognosis in patients with severe
head injury
29-Oct-13

Dr.Rittu Chandel

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CSF phenylalanine
CSF copper

 Total cell count –

Normal - 0 – 5 cells/ μl
Neonate – 0 -30 cells/ μl
No RBC
 Differential count
Lymphocytes: monocytes = 70:30
Neutrophils – 7%
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Dr.Rittu Chandel

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Lumbar puncture findings
test

bacterial

viral

fungal

tuberculous

Opening
pressure

elevated

Usually normal

variable

variable

Leukocyte count > Or =1000/μl

<100/μl

variable

variable

Differential
count

Mainly
neutrophils

Mainly
lymphocytes

Mainly
lymphocytes

Mainly
lymphocytes

protein

Mid – marked
increase

Normal – mild
increase

increased

increased

glucose

< or = 40 mg/dl

normal

decreased

Decreased ;<45
mg/dl

CSF/serum
glucose ratio

Normal –
marked
decrease

Usually normal

low

low

29-Oct-13

Dr.Rittu Chandel

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CSF enzymes
CSF enzymes

disease

Adenosine deaminase

Tubercular meningitis

Creatine kinase ( CK – BB )

Hydrocephalus, cerebral infarction,
primary brain tumors, subarachnoid
hemorrhage

LDH

CNS leukaemia, lymphoma, metastatic
carcinoma, bacterial meningitis, SAH

lysozyme

Bacterial and tuberculous meningitis

Aspartate transaminase

Cerebral abscess, cerebral hemorrhage,
primary or metastatic malignancy

Placental alkaline phosphatase

Germ cell tumor

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Dr.Rittu Chandel

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CSF tumor markers
Carcinoembryonic antigen

Metastatic carcinoma of leptomeninges

Human chorionic gonadotropin

Choriocarcinoma, malignant germ cell
tumors

Alpha feto protein

Germ cell tumors

CSF ferritin

CNS malignancy

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Dr.Rittu Chandel

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Synovial fluid

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Dr.Rittu Chandel

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resilience
Surrounded by shell of water
molecules
A slippery consistency
Compressed – occupy smaller
volume
Relaxed – occupy

29-Oct-13

Dr.Rittu Chandel

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functions
 Reduce friction between bones
 Lubricates joints
 Fluid provides nutrients to cartilage
 Lessens shock of walking and jogging impact

29-Oct-13

Dr.Rittu Chandel

30
Composition
Volume
Color
Clarity

<3.5 mL
pale yellow
clear

Neutrophils
<20% of diff.
Lymphocytes
<15 % of diff.
Monocytes & macrophages
65% of diff.
Crystals
NONE
Glucose
<10 mg/dL (serum synovial
difference)
Lactate
<250 mg/dL
Total protein
<3 g/dL
Uric acid
= blood value
29-Oct-13

Dr.Rittu Chandel

31
collection
*Avoid all powdered anticoagulants – interfere
with crystal analysis
 Fluid verification


29-Oct-13

Mucin clot test Add fluid to dilute acetic acid 
turbidity (clot formation) due to
hyaluronate
Dr.Rittu Chandel

32
examination
Synovial
fluid

Gross

microbiology
29-Oct-13

pathology

Dr.Rittu Chandel

biochemistry
33
Gross examination
 Total volume – recorded at bedside
 Normal= 3.5 mL
 Diseased / inflamed = up to 25

mL

 Colour – colourless

xanthochromia
red brown
 Clarity –transparent
opaque
oily
floating rice bodies
ground pepper appearance
29-Oct-13

Dr.Rittu Chandel

34
Biochemical analysis
 Mucin clot test

Non specific finding
 Glucose
Serum synovial difference < 10 mg/dl
Septic arthritis – difference 20 – 60 mg/dl
 Protein
Normal – 1-3 g/dl
Increased in inflammation
 Uric acid
 RF
 ANA
29-Oct-13

Dr.Rittu Chandel

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Cell count
 Total count

Normal – 150 – 200 / μl
abnormal >10,000/ μl
 DLC
Normal - Neutrophils – 20%
Gout >50%
Acute bacterial arthritis – 75%

29-Oct-13

Dr.Rittu Chandel

36
Differential count






Normal
Neutrophils – 20%
Lymphocytes – 15%
Monocytes and macrophages – 65%
Abnormal
Reiters cell
Eosinophilia >2%

 Microscopic examination
29-Oct-13

Dr.Rittu Chandel

37
Uric acid crystals

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Dr.Rittu Chandel

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crystals

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Dr.Rittu Chandel

39
lymph
 The lymphatic system is an extensive vascular network
 responsible for the transport of fluid , immune cells ,

proteins and lipids .
 The failure to transport lymph fluid results in a
number of disorders and diseases .
 Lymphedema , for example , is a pathology
characterized by the retention of fluid in limbs
creating extreme discomfort , reduced mobility.

29-Oct-13

Dr.Rittu Chandel

40
 slightly basic fluid
 Resembles plasma in its content
 Protein content varies widely from 2% to 8%

29-Oct-13

Dr.Rittu Chandel

41
bibliography
Harrisons internal medicine
Ranna shinde
Vasudevan
Henrys clinical diagnosis

THANK YOU
29-Oct-13

Dr.Rittu Chandel

42

Fluids

  • 1.
    DR.RITTU CHANDEL MD BIOCHEMISTRY(SECOND YR) GRANT GOVT MEDICAL COLLEGE, BYCULLA, MUMBAI – 400008 29 – 10 -2013
  • 2.
    Cerebrospinal fluid Total volumes: –Adults: 140 - 170 ml – Children: 29-Oct-13 cerebral ventricles – 30ml subarachnoid space - 120 ml 10 - 60 ml Dr.Rittu Chandel 2
  • 3.
  • 4.
    Functions of CSF Buoyancy 2.Protective effect 3. Metabolic 4. homeostasis 1. 29-Oct-13 Dr.Rittu Chandel 4
  • 5.
    Specimen collection Cisternal (occipital) Inparalysis Ventricular puncture (lateral occipital) For infants 29-Oct-13 Dr.Rittu Chandel 5
  • 6.
  • 7.
  • 8.
    composition • • • • • • • • Colour - Colourless PH - 7.28– 7.32 Appearance - Clear Sp. Gravity - 1.003 – 1.004 No clot formation on standing Proteins - 10 – 45 mg/dl Glucose - 45 – 100 mg/dl Chlorides - 120 -130 mEq/l 29-Oct-13 Dr.Rittu Chandel 8
  • 9.
           Urea Uric acid Creatinine Cholesterol Ammonia A:G ratio Serum:CSFprotein 29-Oct-13 Dr.Rittu Chandel - 6.0 - 16 mg/dL - 0.5 - 3.0 mg/dL - 0.6 - 1.2 mg/dL - 0.2 - 0.6 mg/dL - 10 – 35 μg/dL - 3:1 – 200:1 9
  • 10.
    Gross examination  Appearance Normal– crystal clear, colourless and no coagulum 1.Colour Any change in colour is always pathological Red Yellow 29-Oct-13 Dr.Rittu Chandel 10
  • 11.
    xanthochromia  Pale pinkto yellow colour in supernatant of CSF Froin’s syndrome 29-Oct-13 Dr.Rittu Chandel 11
  • 12.
    D/D of bloodyCSF  Traumatic tap - hemorrhagic fluid clears between first and third tubes; remains uniform in SAH  Pathological - RBC have crenated appearance  Xanthochromia, erythrophagocytosis and hemosedrin laden macrophages indicate a subarachnoid bleed in absence of prior trauma 29-Oct-13 Dr.Rittu Chandel 12
  • 13.
    Gross examination(contd)  2.Turbidity o WBC >200 cell/μl o RBC >400 cell/μl o Microorganisms, radiographic contrasts, aspirated epidural fat o Protein > 150 mg/dl  Clot formation  Viscous CSF 29-Oct-13 Dr.Rittu Chandel 13
  • 14.
    Pressure of CSF Normal Adults– 90 – 180 mm water Infants and children – 10 – 100 mm water >250 mm water Intracranial hypertension Tumors Decreased pressure Dehydration Circulatory collapse CSF leakage 29-Oct-13 Dr.Rittu Chandel 14
  • 15.
    Biochemical analysis of CSF 1. Glucose Normal – 45 – 100 mg/100 ml decreased – coccal meningitis TB meningitis intrathecal streptomycin Increased – diabetic hyperglycemia In assessing response to treatment 29-Oct-13 Dr.Rittu Chandel 15
  • 16.
     2.Proteins Normal –15 – 45 mg/dl Term infants – 90 – 150 mg/dl Pre term - 115 – 170 mg/dl  Increased CSF protein I. Increased permeability of blood brain barrier II. Increased intrathecal IgG secretion 29-Oct-13 Dr.Rittu Chandel 16
  • 17.
  • 18.
  • 19.
    Permeability of bloodbrain barrier CSF/serum albumin index index permeability <9 intact barrier 9 – 14 slight impairment 14 – 30 moderate impairment >30 severe impairment Index slightly elevated in infants up to 6 months of age 29-Oct-13 Dr.Rittu Chandel 19
  • 20.
    CSF serum immunoglobulin ratio Increases in multiple sclerosis  Increased CSF IgM and kappa light chains – marker of multiple sclerosis 29-Oct-13 Dr.Rittu Chandel 20
  • 21.
    CSF proteins andCNS diseases proteins Diseases or disorders Α2 - macroglobulin Subdural hemorrhage, bacterial meningitis Β – amyloid protein 42 and ζ protein Alzheimer’s disease Β 2 - microglobulin Leukemia/lymphoma, bechets syndrome C- reactive protein Bacterial and viral meningitis fibronectin Lymphoblastic leukemia, AIDS, meningitis methemoglobin Mild subarachnoid/subdural hemorrhage Mylein basic protein Multiple sclerosis, tumors Protein 14- 3 -3 29-Oct-13 Dr.Rittu Chandel Creutzfeldt – jacob disease 21
  • 22.
  • 23.
    CSF lactate  CSFand blood lactate are largely independent  Normal newborn – 10 – 60 mg/dl Child and adult – 9 – 26 mg/dl Differentiating meningitis CSF lactate (mg/dl) meningitis < 25 mg/dl Almost always less than 35 mg/dl viral >35 mg/dl bacterial Persistently elevated - poor prognosis in patients with severe head injury 29-Oct-13 Dr.Rittu Chandel 23
  • 24.
    CSF phenylalanine CSF copper Total cell count – Normal - 0 – 5 cells/ μl Neonate – 0 -30 cells/ μl No RBC  Differential count Lymphocytes: monocytes = 70:30 Neutrophils – 7% 29-Oct-13 Dr.Rittu Chandel 24
  • 25.
    Lumbar puncture findings test bacterial viral fungal tuberculous Opening pressure elevated Usuallynormal variable variable Leukocyte count > Or =1000/μl <100/μl variable variable Differential count Mainly neutrophils Mainly lymphocytes Mainly lymphocytes Mainly lymphocytes protein Mid – marked increase Normal – mild increase increased increased glucose < or = 40 mg/dl normal decreased Decreased ;<45 mg/dl CSF/serum glucose ratio Normal – marked decrease Usually normal low low 29-Oct-13 Dr.Rittu Chandel 25
  • 26.
    CSF enzymes CSF enzymes disease Adenosinedeaminase Tubercular meningitis Creatine kinase ( CK – BB ) Hydrocephalus, cerebral infarction, primary brain tumors, subarachnoid hemorrhage LDH CNS leukaemia, lymphoma, metastatic carcinoma, bacterial meningitis, SAH lysozyme Bacterial and tuberculous meningitis Aspartate transaminase Cerebral abscess, cerebral hemorrhage, primary or metastatic malignancy Placental alkaline phosphatase Germ cell tumor 29-Oct-13 Dr.Rittu Chandel 26
  • 27.
    CSF tumor markers Carcinoembryonicantigen Metastatic carcinoma of leptomeninges Human chorionic gonadotropin Choriocarcinoma, malignant germ cell tumors Alpha feto protein Germ cell tumors CSF ferritin CNS malignancy 29-Oct-13 Dr.Rittu Chandel 27
  • 28.
  • 29.
    resilience Surrounded by shellof water molecules A slippery consistency Compressed – occupy smaller volume Relaxed – occupy 29-Oct-13 Dr.Rittu Chandel 29
  • 30.
    functions  Reduce frictionbetween bones  Lubricates joints  Fluid provides nutrients to cartilage  Lessens shock of walking and jogging impact 29-Oct-13 Dr.Rittu Chandel 30
  • 31.
    Composition Volume Color Clarity <3.5 mL pale yellow clear Neutrophils <20%of diff. Lymphocytes <15 % of diff. Monocytes & macrophages 65% of diff. Crystals NONE Glucose <10 mg/dL (serum synovial difference) Lactate <250 mg/dL Total protein <3 g/dL Uric acid = blood value 29-Oct-13 Dr.Rittu Chandel 31
  • 32.
    collection *Avoid all powderedanticoagulants – interfere with crystal analysis  Fluid verification  29-Oct-13 Mucin clot test Add fluid to dilute acetic acid  turbidity (clot formation) due to hyaluronate Dr.Rittu Chandel 32
  • 33.
  • 34.
    Gross examination  Totalvolume – recorded at bedside  Normal= 3.5 mL  Diseased / inflamed = up to 25 mL  Colour – colourless xanthochromia red brown  Clarity –transparent opaque oily floating rice bodies ground pepper appearance 29-Oct-13 Dr.Rittu Chandel 34
  • 35.
    Biochemical analysis  Mucinclot test Non specific finding  Glucose Serum synovial difference < 10 mg/dl Septic arthritis – difference 20 – 60 mg/dl  Protein Normal – 1-3 g/dl Increased in inflammation  Uric acid  RF  ANA 29-Oct-13 Dr.Rittu Chandel 35
  • 36.
    Cell count  Totalcount Normal – 150 – 200 / μl abnormal >10,000/ μl  DLC Normal - Neutrophils – 20% Gout >50% Acute bacterial arthritis – 75% 29-Oct-13 Dr.Rittu Chandel 36
  • 37.
    Differential count      Normal Neutrophils –20% Lymphocytes – 15% Monocytes and macrophages – 65% Abnormal Reiters cell Eosinophilia >2%  Microscopic examination 29-Oct-13 Dr.Rittu Chandel 37
  • 38.
  • 39.
  • 40.
    lymph  The lymphaticsystem is an extensive vascular network  responsible for the transport of fluid , immune cells , proteins and lipids .  The failure to transport lymph fluid results in a number of disorders and diseases .  Lymphedema , for example , is a pathology characterized by the retention of fluid in limbs creating extreme discomfort , reduced mobility. 29-Oct-13 Dr.Rittu Chandel 40
  • 41.
     slightly basicfluid  Resembles plasma in its content  Protein content varies widely from 2% to 8% 29-Oct-13 Dr.Rittu Chandel 41
  • 42.
    bibliography Harrisons internal medicine Rannashinde Vasudevan Henrys clinical diagnosis THANK YOU 29-Oct-13 Dr.Rittu Chandel 42