Urinalysis and Body Fluids CRg

Student review for Competency and BF Lab Practical

Neubauer hemacytometer / counting chamber• Large squares, such as # 1-9 below have volume of 0.1• Small squares – in center # 5 have volume of 0.004

Improved Neubauer hemacytometer / counting chamber• Counting the cells

• Cells touching either the top or left boundary are included in the count

• When triple lines are present, the MIDDLE line is used as the boundary.

Neubauer hemacytometer / counting chamber• Formula for calculations – results in # cells / uL

• Count and record cells from both sides of the chamber.• Average the two sides• Multiply by dilution factor (if no dilution is made, this number is 1)• Divide by number of squares counted X volume of each square

• Large squares, such as # 1-9 below have volume of 0.1• Small squares – in center # 5 have volume of 0.004

ave . # ce l ls counted x d ilution# squares counted x vo lume of each square

Body Fluid Differential• Specimens for cell differential – usually collected in

EDTA tube (except CSF samples)

• Cytospin preparations produce more accurate counts, however cell distortion often seen (can be minimized by adding albumin to sample before processing).

• Stain = Wright/Wright-Giemsa (Gram stain may be used if bacterial infection suspected.)

• Hyaluronidase added to synovial fluid to reduce the viscosity and make the slide more uniform.

Body Fluid Differential

• Any nucleated peripheral blood cell, including immature forms, plasma, and LE cells can be found.• Check textbook / classroom notes for normal values.

• Monocytes/macrophages with inclusions are renamed• Erythrophages contain one or more red cells• Siderophages contain hemosiderin /siderotic bodies or

hematin crystals• Leukophages contain white blood cell(s)• Lipophage contains ingested fat/lipid

Body Fluid Differential

• Entire smear should be evaluated for• abnormal cells• inclusions within cells• Clusters• Presence of intracellular organisms

• CSF specimen cells• L – neutrophiles & @6 macrophages

• R – neutrophiles and intracellular bacteria

Body Fluid Differential

Body Fluid Differential - CSF

• Eosinophils• Often associated with parasitic / fungal infections,

allergic reactions including reaction to shunts and other foreign objects.

• Ependymal cells• Normal cell, unique to CSF • Line the ventricles, produce CSF fluid• Large cell with distinct round/oval

nucleus, sometimes found in sheets

Body Fluid Differential - CSF

• Cellular inclusions• Erythrophage• Siderophage• Hematoidin crystals (see below)

•ASCP 21 CSF erythrophage, with few iron granules forming

ASCP 6 macrophage, lymphocyte, siderophage

1991 CAP 30 CSF hematoidin crystal / bilirubin crystal

Body Fluid Differential - CSF

• India-ink / nigrosin preparation • Negative stain to view the encapsulated

Cryptococcus neoformans (frequent complication seen in AIDs /immunocompromised patients)

• Instead of using stain, can also use dark field microscopy to get same effect.

• Has @ 25-50% sensitivity • Serological latex agglutination tests provide better

results – and are preferred.

Body Fluid Differential - CSF

• Suspicious / unclassified or malignant cells are reported as “other” or “unclassified” AND are sent to pathology. (as seen below)

• Cytology – send unstained slide to cytology / pathology• 1986 CAP CM10 CSF – blasts (appearance similar to peripheral blood,

always consult with hematology specialist / pathologist) ( see below right)

Body Fluid Differential

• left is 1988 CAP CM 25 – CSF 250x malignant cells• Remember – we classify them as ‘other’ or ‘unclassified’

and take the slide to the cytologist / pathologist• Right is leukemic cells found in CSF

Body Fluid Differential

• Serous fluid examples• Left -lymphocyte, macrophage/mono, basophil• Right - Mesothelial cells

Body Fluid Differential - Serous

Serous fluid - Mesothelial cell

• Mesothelial cell

Serous Fluids• Malignant cells• ACSP 7 Case 1 cont.

peritoneal fluid, malignancy

Serous Fluids• Malignant cells• ASCP 12 Case 4, 30 year old

with back pain and inability to work. Pleural effusion fluid – malignant tumor on spinal cord

Serous Fluids-LE cells • Seen in patients with Systemic Lupus Erythmatosis (SLE) a

systemic disease in which an autoantibody attacks the patients organs and body systems

• LE cell is a neutrophil or macrophage that has engulfed a homogeneous mass of purple staining nuclear material

•It is not unusual to find macrophages engulfing other cells,. If the nucleus of the ingested cell has intact nuclear structures / normal chromatin pattern, the macrophage is considered to be a Tart cell . Tart cells are not associated with SLE.•Example is a segmented neutrophil and demonstrates the LE cell characteristic of containing a smooth homogenous nuclear mass.

Synovial Fluid • In addition to counting and

identifying cells; identification of any crystals present aids in diagnosis.

•Monosodium urate associated with gouty arthritis.

•Calcium pyrophosphate associated with pseudogout.

•Blunt ended rod-like crystals resembling monosodium urate, crystals can be seen in patients following joint steroid injection.

•Others: oxalate and cholesterol

The Gout

• 1799 caricature by James Gillray