Important Extra Information Doctors slides Doctors notes " ما تعثرت ارتباكا يا حصى .. او جنوحا لإختصار الالف ميل  ليس ذنبي أن دربي ما استوى..  كان ذنبي ثوب احلامي طويل! " جلال الدين الرومي Biochemistry Cerebrospinal Fluid (CSF) Analysis for total protein

CSF Examination Report: Physical examination: Color Appearance Viscosity Chemical examination: CSF protein concentration (g/L) Group number & Student names.

Indications for CSF analysis CSF examination Indications for CSF analysis CNS infections Demyelinating diseases malignancy Hemorrhage in CNS Autoimmune disorders The following examinations are performed for CNS samples: Physical examination Microscopic examination Chemical examination Microbiological test

1- Physical Examination: Parameter Normal abnormal Volume 100-150 ml (8% of CNS cavity volume) Turbidity Clear Cloudy/turbid- may indicate the presence of: white, or red blood cells, microorganisms, or an increase in protein level. Color Colorless Changes in color may point to additional substances in the fluid. Yellow (Xanthochromia)1, orange-brown or red may indicate the presence of blood or hemoglobin pigments2. Viscosity3 same consistency as water Thicker CSF may be seen in patients with certain types of cancers or meningitis. Xanthochromia is the yellow discoloration indicating the presence of bilirubin in the (CSF). The two most common reasons for blood and hemoglobin pigments in CSF are: traumatic tap (color is bright and red and erythrocytes number is decreased. subarachnoid hemorrhage (SAH) = xanthochromia is present. you should shake the sample to inspect its viscosity.

2- Chemical Analysis: Remember !! Centrifuge: Before any analysis, the fluid should be centrifuged to avoid contamination by cellular elements. CSF is the most precious biological material. Often, only small volumes of CSF are available for analysis due to difficulty in collection; hence handle this with care. The specimen may not be pre-tested for micro-organisms ( may contain virulent organisms), so strict safety precautions should be followed. Routinely performed biochemical tests in CSF are: Total protein (total- specific e.g. albumin & Immunoglobulin) glutamine and acid-base parameters lactate lactate dehydrogenase glucose protein is normally low in CSF: if increased suspect infection, but its decrease is not significant. lactate and lactate dehydrogenase increase in bacterial infection, but not in viral. glutamine increase in CSF due to liver disease

Abnormal findings of CSF in some pathological conditions Mind map Turbidity Physical Examination Color Viscosity CSF Protein Assay Spectrophotometer glucose Cerebrospinal Fluid (CSF) protein (total and specific) Chemical Analysis Procedure Mind map by 432 biochemistry team lactate Calculation lactate dehydrogenase Normal Range Abnormal findings of CSF in some pathological conditions glutamine and acid-base parameters

CSF Protein Assay Spectrophotometer “Extra Information” Protein present in the CSF is detected by a series of enzymatic reactions that ultimately form a colored product like kit based on Biuret method. Biuret reagent when interacts with the peptide bonds in the protein give a blue coloured product. The intensity of the color is proportional the amount of protein in CSF Color intensity is determined by measuring the absorbance by the colored solution at a wavelength of 546nm Absorbance is measured by an instrument, spectrophotometer Spectrophotometer “Extra Information” Most of visible spectrophotometers are composed of: Light source which works with visible wavelengths (400-700 nm) Monochromator filter for choosing desired wavelength Sample holder (cuvette) Detector Meter or recorder

Tuberculous Meningitis Abnormal findings of CSF in some pathological conditions Very important Condition Parameter Brain tumor Viral Meningitis Tuberculous Meningitis Bacterial Meningitis (pyogenic) ↑ Normal ↑↑ Proteins ↓ Normal or slightly ↓ ↓↓ Glucose Normal or ↓ Chloride

Normal reference values for CSF protein: Procedure Normal Range Normal reference values for CSF protein: 15 - 45 mg/dL ( 0.1 - 0.4 g/L ) Test Standard Blank Reagent 2 ml CSF sample 40 l - Dist.H2O Mix and incubate for 15 minutes at room temperature Measure absorbance at 546 nm Calculation Protein concentration (g/L) = Absorbance of sample ”test” X Conc. of standard (60 g/L) Absorbance of standard Important: you must convert from mg/dL to g/L ) , divide on 100 .

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