1. Urinalysis and Body Fluids CRg
Student review
for Competency and BF Lab Practical

2. 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
RBC and WBC counts are reported as # cells / uL
To obtain this you must Count and record cells from both sides of the chamber.
Average the two sides
And enter this information into the formula shown
The average number of cells from both sides of the chamber are multiplied by the dilution factor . This number will be 1 ,unless you needed to make a dilution. (
On the denominator of the calculation, you enter the number of squares you counted each side of the chamber. Times the volume of each square.
Large squares, such as # 1-9 seen on the picture below have volume of 0.1
Small squares – in center # 5 have volume of 0.004

3. 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.

4. 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

5. 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.

6. 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

7. Body Fluid Differential
Entire smear should be evaluated for
abnormal cells
inclusions within cells
Clusters
Presence of intracellular organisms

8. Body Fluid Differential
CSF specimen cells
L – neutrophiles macrophages
R – neutrophiles and intracellular bacteria

9. Body Fluid Differential - CSF
Eosinophils
Often associated with parasitic / fungal infections, allergic reactions including reaction to shunts and other foreign objects.

10. Body Fluid Differential - CSF
Ependymal cells
Normal cell, unique to CSF
Line the ventricles, produce CSF fluid
Large cell with distinct round/oval nucleus, sometimes found in sheets

11. Body Fluid Differential - CSF
Cellular inclusions
Erythrophage
Siderophage
Hematoidin crystals (see below)
ASCP 21 CSF erythrophage, with few iron granules forming
1991 CAP 30 CSF hematoidin crystal / bilirubin crystal
ASCP 6 macrophage, lymphocyte, siderophage

12. 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.
% sensitivity
Serological latex agglutination tests provide better results – and are preferred.

13. Body Fluid Differential
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)

14. Body Fluid Differential
left is 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

15. Body Fluid Differential - Serous
Serous fluid examples
Left -lymphocyte, macrophage/mono, basophil
Right - Mesothelial cells

16. Serous fluid - Mesothelial cell

17. Serous Fluids Malignant cells
ACSP 7 Case 1 cont. peritoneal fluid, malignancy

18. 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

19. 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.

20. 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

21. The Gout
1799 caricature by James Gillray