Week 7 Flashcards
how do you obtain a csf
lumbar puncture
how do you obtain a serous fluid
needle aspiration
Thoracentesis
Pericardiocentesis
paracentesis
how do you obtain synovial fluid
needle aspiration
arthrocentesis
do we anti coagulate the samples or no
cells counts cant be done on clotted samples
-Csf doesnt have fibrinogen since the BBB controls filtration of plasma compenents
Serous fluids and synovial fluids are plasma ultra filtrates
seminal fluid clots and liquefies
What type tube does each fluid need
CSF- 2-3 ml, 3-4 tube with NO anticoagulant
Serous - ~100 ml in EDTA
Synovial fluid - 3 tubes in EDTA or heparin
seminal fluid - 2-3ml in sterile container no anticoagulant
What do we look at the neat fluid for and what does it indicate
Colour
Turbidity
Consistency
abnormality type and how its processed
if its red or bloody
yellow or xanthochromic
be blood or hemolysis - centrifuge and look at SUPERNATANT and then describe the supernatant color
bilirubin
if fluid is turbid what does it indicate
infection
what are the types of consistency the sample can have
Stringy
Clotted
Thick
testing times for fluids
CSF - in one hour
Others - 24 hours
when you look at the BF what type of processing options are there
counting manually not diluted
manual counting with dilution
Manuals must be done for low CSF counts
automated on cell counter - sample must be bloody, must check analyzer linearity because they may not be accurate at low RBC counts . Not good for WBC
How do you do a manual count of a clear fluid
-add to chamber neat or undiluted
-entire chamber is counted (9 squares on both sides)
how do you count cloudy , blood fluids
-dilute 1:10 or 1:200
-count all 9 squares if there arent too many cells
- do WBC squares or RBC squares
how to dilute samples for a low count
-use the smallest dilution
-flood chamber with neat sample and assess
-less accuracy with dilution
-use saline for RBC/WBC
-acetic acid to lyse RBC = for WBC
-TURK solution with acetic acid and methylene blue for WBC count since it stains nuclei
-DONT USE ACETIC ACID FOR SYNOVIAL FLUID
ADD HYALURONIDASE to synovial fluid to liquefy before doing counts or prepping cytocentrifuge slides
What is the cytocentrifuge
how are they prepped
concentrates sample into a small area so counts can be done
-you can also spin sample and make a circular sediment smear
Must have a circle on slide to find cells – low cell counts are hard to see
-stain with Wrights stain
-count 100 wbc and nrbc
-comment on bacteria that are present
What are the normal lining cells in the
CSF
SEROUS FLUID
SYNOVIAL FLUID
-Ependymal & choroid plexus cells, spindle-shaped cells, Large with lots of cytoplasm, found in clumps
-Mesothelial cells
-Synoviocytes (synovial lining cells)
How are lining cells different than normal ones
Lining cells are :
- larger
- flat clumps or sheets
- large, central, round nuclei, multi-nuclei
- moderately abundant blue cytoplasm
What cells are seen in
normal
Bacterial infection
Viral infection
Parasitic infections
Allergic reaction
Shunt
normal- hypersegmented neuts with filaments , few lymphs in adults and monocytes for peds,
Bacterial infection - neutrophils (meningitis) early TB, viral or fungal
Viral infection - lymphs, and reactive lymphs
Parasitic infections - eos, basos
Allergic reaction
Shunt
What cells are seen in fluid of
Leukemia /Lymphoma
Inflammation
TRAUMA
Caner/tumors
Leukemia /Lymphoma - Leukemic Blasts, Lymphoma cells, Plasma cells
Inflammation -Neutrophils, LE cells
TRAUMA -Red Blood Cells Siderophages
Caner/tumors -Tumour cells
recognition of these cells is done in diagnostic cytology
-send smear to pathologist
when looking at a CSF
gross appearance
Clear = report
Cloudy, bloody - spin and report supernatant appearance
Cell count
Clear - count UNDILUTED
cloudy/bloody - dilute
Turks (1:2 or WBC 1:20 = WBC reported
RBC 1:200 - report RBC, fresh and crenated
diseased CSF can cause
how many tubes and which goes to what department
hydrocephalus - increased volume
Meningitis - symptoms of headache, fever
trauma can cause intracranial hemorrhage
STAT= draw 3-4 tubes -No Anticoagulant
analyze now since RBC and WBC can degenerate
Tube # 1 for Chemistry & serology 2-8 fridge
Tube # 2 for Microbiology - RT (keep orgs viable)
Tube # 3 for Hematology -2-8 fridge
Cell counts
Tube # 4 (if possible) for
cytology
What do we look for in a visual assessment
-all tubes assessed first
-turbid, cloudy, = meningitis , hemorrhage, blood tap
-bloody = hemorrhage= blood tap
-pellicle = TB
What are the 2 possibilities when assessing a blood CSF
-intracranial bleeding due to trauma
-blood in sample only because of poor procurement technique (bloody tap)
What must hematology do before all departments
- look at all tubes if bloody
-count cells BEFORE chem spins
-enter Micro tube with sterile technique
-smears or cytospins on all tubes
-send rest to all departments
What are the characteristics of a traumatic tap
Clear supernatant
Clearing from tube to tube
1st tube cloudiest
Progressively less cloudy
Fresh RBCs
What are the characteristics of a Pathological Hemorrhage
Xanthochromic / hemolyzed supernatant
Same appearance in all tubes
Crenated RBCs
a blood tap get progressively less bloody whereas hemorrhage is bloody in all tubes
when spun a blood tap has a clear supernatant whereas a hemorrhage has a supernatant which is xanthochromic due bilirubin from broken cells
blood from blood tap may clot
What does CSF dilution in saline depend on
appearance
Clear —- Undiluted
Hazy —– 1/10
Cloudy —1/100
Bloody/Turbid —–1/1000
What is the 1 count method for CSF
load chamber with neat CSF on both sides
-count all cells that differentiating WBC and RBC
-dont diff the WBC in a counting chamber
-no dilution
What is the 2 Dilutions/counts
-if you have a higher cell count do TWO different cell counts Total cell count and WBC count
-count all cells without dilution = TOTAL cell count
-dilute with acetic acid - WBC only since RBC got lysed
Total - WBC = RBC so you dont have differentiate the two types but you still do 2 types of assessments
What is the QC required on CSF
needed to assess the count
check equipment
check for reagent contamination
incorrect concentration of acetic acid
When must you do cytospin on csf
if any WBC are seen
commonly seen orgs on a gram of csf
GNDC
GPC
GNCB - H FLU
Crypto
Where do you take serous fluids from
and when can you aspirate them
- they are lubricant in small closed cavities
pleural - round the lung
pericardial - around the heart
peritoneal - around the intestine
-only take if there is a disease process
constant volume
Production = reabsorption
What is Effusion in serous fluids
-when fluid accumulates in a cavity
ascites = large amount of fluid in peritoneal cavity
divided into transudates and exudates
What is a transudate
-problem in formation/absorption mechanism
-formed due to increased capillary hydrostatic pressure or decreased plasma oncotic pressure
-develop as part of system disease process
-seen in congestive heart failure
What are exudates
-problem with membrane
-problem with reabsorption at the location where the fluid originates
-also occur due to increased capillary permeability or decreased lymphatic resorption
-indicates inflammatory disorders that are associated with bacterial/viral infections, malignancy, pulmonary emboli, and SLE
if a serous effusion is turbid and cloudy what does it suggest depending on where its from
Pleural: microbial infection, pneumonia, TB
Pericardial: Bacterial endocarditis
Peritoneal: Peritonitis (perforated bowel, appendix)
If fluid is cloudy dilute with Turk’s fluid: Count Total or RBC then WBC with Turk’s
why would a serous effusion be milky
chylous material (triglycerides) from thoracic duct
Why would a serous effusion be bloody
Hemothorax – hemorrhage in pleural sac
What are cells that are normally seen in serous fluids
Mesothelial cells that look like fried eggs
-lymphs
-macros
What are some abnormal cells seen in serous fluids
-malignant cells from primary or metastatic tumors
-same as the malignant cells found in CSF
-neuts in bacterial infection
-Eos and basos due to allergic sensitivity of foreign material
What is seen in in pleural cells due to SLE
LE cell
-appear due to environmental conditions
AB in fluid, Trauma to cells, incubation
-ANA or RA done on pleural fluids
synovial fluid
what is it
what does it secrete
-formed as ultrafiltrate of plasma
-normal is 3.5 ml
-membrane cells secrete Hyraluronic acid to make the fluid viscous and allow for joint movement
how is synovial fluid tested
- can clot so anticoagulant can be used
-use EDTA or heparinized tube
-normally it will be viscous with few cells and no crystals
do a visual check with color, turbidity, viscosity, and microscopic examination for crystals and cells
how is viscosity assessed for synovial fluids
-measure ability of the fluid to form a string 4-6 cm long
-hyaluronic acid helps with joint movement
-makes cell counts smear prep difficult
-to reduce viscosity add ‘Hyaluronidase’ enzyme to liquefy the fluid AFTER you look for crystals
What types of crystals can you see in synovial joint fluid
Monosodium urate - NEEDLE LIKE
Calcium Pyro - RHOMBOID
Cholesterol - NOTCHED WEDGE
What are the different types of joint disorders
Noninflammatory - degenerative disorder
Inflammatory /Immunological - RA, SLE, = crystal induced: Gout or Pseudogout
Septic - Microbial infection
Hemorrhagic -Trauma, Bleeding disorders
What can different colored fluid suggest in synovial fluid abnormalities
Deeper yellow with inflammation
Green-yellow with infection
Red : Presence of blood due to hemorrhage or traumatic tap
Brown/Pink : Old hemorrhage / lysed blood
how are cell counts done with synovial fluid
same way as CSF
-higher dilution is used for turbid samples
-count cells in 5 large squares and multiply by dilution to get cells/uL
-WBCS will be high in inflammatory, septic and hemorrahagic joint disorders
-RBCs are increased in hemorrhagic disorders
What are normal cells seen in synovial fluid
-lymphocytes, Monocytes/histocytes
-synovial cells are lining cells that look like mesothelial cells
How do you prep for looking at crystals from a fluid
-can have intra or extracellular crystals
-can view on slide with coverslip or make a cytospin
-examin with polarizing mic and red filter
What can crystal in fluid result in
what are the most common types and causes
-result in acute, painful inflammation
common
Monosodium urate (MSU) in gout
Calcium pyrophosphate (CPPD) in pseudogout- stain with Wrights (blue when parallel to polarized light and yellow when rotated 90 degrees)
Cholesterol in chronic arthritis and rheumatoid arthritis
Caused by
Metabolic disorders
Decreased renal excretion
Increased intake
Degeneration of cartilage and bone
Injection of medication viz. corticosteroids
What will you see in non inflammatory disorders in synovial fluid
Colour/Clarity: Yellow/clear
Good viscosity
WBCs <3000/µL, Neutrophils<30%
RBCs not present
What will you see in non Inflammatory Disorders
(Immunologic) in synovial fluid
Colour/Clarity: Yellow/Cloudy
Poor viscosity
WBCs 3000 – 75,000 /µL, Neutrophils >50%
RBCs not present
What will you see in Inflammatory
(Crystal-induced) in synovial fluid
Colour/Clarity: Yellow/Cloudy, milky fluid
Poor Viscosity
WBCs 500 – 200,000 /µL, Neutrophils <90%
RBCs not present
What will you see in Septic Disorders
in synovial fluid
Colour/Clarity: yellow-green/turbid
Poor viscosity
WBCs 50,000-200,000 /µL, Neutrophils >90%
RBCs present
What will you see in Hemorrhagic Disorders
in synovial fluid
Colour/Clarity: Red-brown/cloudy
Poor viscosity
WBCs 50 – 10,000/µL, Neutrophils <50%
RBCs >2000/µL
What will you see in Autoimmune
in synovial fluid
LE cells in SLE & ANA positive in SLE
RA factor in RA
What do we look at seminal fluid for
infertility
post vasectomy
semen has four fractions
Testes and epididymis: Sperms (5%)
Seminal vesicles: Fluid rich in fructose (60%)
Prostate: Acids and proteolytic enzymes (30%)
Bulbourethral glands: Alkaline mucus (5%)
What is assesed in semen
appearance - GREY TRANSLUCENT
-volume 2-5ML
-viscosity
-sperm count
-sperm concentration (density)- # per volume
-sperm motility >50% IN AN HOUR
-% of normal and abnormal (defective)
-coagulation and liquefaction - thick to liquid (POUR IN DROPLETS AND LIQUID IN 30-60 MINS)
-fructose = energy
pH = acidity SHOULD BE 7.2-8
-immature sperm
-# of WBCs
how is sperm collected
collected by masturbation after >3 days and <5 days of sexual abstinence
-WHOLE specimen needs to be sent to the lab within an hour of collection
-note collection time
-keep at 37C until liquefaction then process immediately
how do we study sperm morph
-thin smear
-air dry
-Wright’s, Giemsa or Papanicolaou stain
-count 200 sperm
-look for abnormal forms
-Note WBCs or Premature sperms if present
how to count sperm
1/20 dilution in fluid/saline
-charge chamber
-count sperms in 5 RBC squares
Sperms/mL = Sperms counted X 106
Total Sperm Count = Sperms/mL X Vol of Fluid
What tested post vasectomy
- confirm success of vasectomy by absence of spermatozoa
-do monthly intervals until 2 consecutive specimens show no sperms
-look for sperm on wet prep
-motile or non motile
-confirm absence by spinning for 10 mins and looking at sediment
