Clinical Haematology Flashcards
what are the erythrocyte (RBC) parameters shown on a haemogram?
manual reticulocyte count haemaglobin haematocrit number of RBC mean cell volume mean cell heamoglobin mean cell heamoglobin concentration
what value on a haemogram is most often wrong?
platelets
what are the main leukocyte (WBC) parameters on a haemogram?
total WBC neutrophils lymphocytes monocytes eosinophils basophils
what is the best tube to used for haematology?
EDTA blood tube
what does EDTA do to blood in the tube?
chelates (binds) calcium in the blood. This is required for clotting so EDTA prevents clotting.
when may heparin tubes be used?
in some exotic species as EDTA can cause lysis
what is essential when filling tubes for haematology?
respect the amount of blood required in the tube - only fill to the line
what must you do to haematology tubes to ensure good mixing of blood with anti-coagulate?
gently invert the tube 10-20 times and roll
how should blood smears be stored?
once dry - in slide containers
why should blood smears not be stored in the fridge?
condenses the cells and leads to water artefact
where should haematology samples be stored?
in the fridge until submission to the lab or running sample
why should haematology samples not be stored in the freezer?
causes cell rupture
how should haematology samples be packaged?
not right next to the ice pack as this may freeze them
what parameters evaluate RBC?
haematocrit
packed cell volume
RBC count
what parameters show average makeup of RBC/indexes?
mean corpuscular volume
mean corpuscular haemoglobin
mean corpuscular haemoglobin concentration
what test is used to assess RBC morphology?
peripheral blood smear exam
what parameters are directly measured by haematology analyser?
haemoglobin
red blood cell count
mean cell volume
what does the mean cell volume show?
average size of RBC
what can be calculated from parameters measured by haematology machine?
haematocrit
mean corpuscular haemoglobin
mean corpuscular haemoglobin concentration
how can haematocrit be calculated?
mean cell volume x red blood cell count
how can mean corpuscular haemoglobin be calculated?
Haemoglobin x 10/red blood cell count
how can mean corpuscular haemoglobin concentration be calculated?
Haemoglobin / haematocrit
what are the two types of haematology analysers?
flow cytometry
impedance
how do flow cytometry haematology analysers work?
individual cells pass through a laser beam absorbing and scattering light. Interruptions in light count cells and light scatter is used to determine size of cell and the internal complexity. Produces differential count
How is cell size measured by flow cytometry?
Interruptions in light count cells
how is cell size and complexity measured by haematology analyser?
light scatter
how do impedance haematology analysers work?
individual cells pass an isotonic solution between two electrodes. Cells are poor electrical conductors so when passed between electrodes they produce a change in electrical impedance that is proportional to the size of the cell
what form of haematology analyser is preferable?
flow cytometry can produce a differential count
what are the 9 common sample artefacts which interfere with automated CBCs?
clots of any size platelet clumps (may be read as a cell duce to lack of nucleus in mammalian RBC) macroplatelets RBC agglutination (RBC with nucleus) nRBC Heinz bodies Lipaemia leukocyte agglutination delay in sample handling (increased haemolysis)
what is packed cell volume?
PCV: percentage of RBC in a volume of blood
How is PCV read?
after centrifugation, the percentage of RBC in column of the capillary tube
what else can be assessed during PCV test?
buffy coat assessment - number of WBC
Plasma colour
total proteins
what does the buffy coat on a PCV test show?
WBC and platelets, should be small
what does plasma colour indicate?
straw/clear = normal
bright yellow = icteric
pink = haemolysed
milky and turbid = lipaemia
why is a blood smear so important?
morphology changes are not picked up by any analysers
will show discrepancies/errors in any analyser (QA)
help with quick clinical decisions
PCV+Blood smear are a low cost option when no machines available
what are the minimum levels of patients that should have their blood smear reviewed?
all critically ill patients
CBCs with unusual or suspicious results
what would indicate the need for performing a blood smear?
presence of:
nucleated RBC (indicating reduced RBC or high need for RBC)
neutrophil left shift (immature neutrophils)
unclassified or unidentified cells
automated WBC count that may not be accurate
what background would trigger a blood smear review?
unusual background matrix
unusual background colour
organisms or suspected organisms
what RBC parameters would trigger at blood smear review?
moderate to marked poikilocytosis of any kind (abnormally shaped RBC)
moderate to sever anaemias
any Heinz bodies in non-feline species (>10% in cats)
inclusions (organisms or suspected organisms)
Howell-Jolly bodies
abnormal MCV
what is poikilocyotsis?
strange shaped cells
what WBC parameters would trigger blood smear review?
left shift (marked or degenerative) leukopenia leukocytosis lymphocytosis unclassified cells organisms (or suspected) presence of granules in non-monocytes and abnormal granulation of any leukocyte
what is leukopenia?
decrease in number of WBC (<3,000 WBC)
what is leukocytosis?
Increase in total WBC count (>30,000 WBC)
what is lymphocytosis?
elevation of lymphocytes (>10,000 cells)
what platelet parameters would trigger blood smear review?
> 900,000 platelet count
thrombocytopenia
abnormal MPV
suspected inclusions or abnormal granulation
what is thrombocytopenia?
low platelets (<100,000 cells)
what are the 3 main parts of a blood smear?
base/head
monolayer
feathered edge
what are the measured areas of a blood smear?
monolayer and feathered edge
how do RBC appear in the monolayer of the blood smear?
side by side, not overlapping
how should you start looking at a blood smear?
small - low magnification and at the feathered edge of the smear
what is the systemic approach to a blood smear exam?
start small at low magnification from the feathered end
go 2-3 fields back to the body of the smear in the monolayer
increase to oil and evaluate morphology
finish at the side, count 100 leukocytes into types looking for abnormal forms as you go
what do you look at regarding RBC in a blood smear?
numbers (does it loo anaemic)
do RBC look normal
is there evidence of regeneration
what do you look at regarding WBC in a blood smear?
number
type present
morphology
what do you look at regarding platelets in a blood smear?
number (estimate)
morphology (size)
what is the function of RBC?
oxygen carrying to tissues
how do RBC appear in dogs?
central pallor taking 1/3 of cell
how do RBC appear in cats?
small
no central pallor
all cells same size/colour
what is the role of neutrophils?
defence against invading microorganisms especially bacteria
when do neutrophils increase?
in inflammation and infection. Stress due to adrenaline and corticosteroids
what do neutrophils look like under a microscope?
ribbon shaped, segmented nucleus
pale cytoplasm
what do eosinophils do?
defence against parasites
allergic response
what do basophils do?
defence against parasites
allergic response as contian histemine
what do basophils do?
defence against parasites
allergic response as contain histamine
what are granulocytes?
collective name for neutrophils, basophils and eosinophils
how do eosinophils and basophils differ under a microscope?
basophils are blue toned. Eosinophils have pink granules with purple nucleus
what are lymphocytes involved in?
immunity both adaptive/cell mediated (T cell-) and humoral (b cells - antibody production)
how do lymphocytes appear under a microscope?
round cells with a large round nucleus
very little cytoplasm
what are monocytes?
precursor of macrophages
what are monocytes involved in?
phagocytosis
antigen presentation to T cells
immunomodulation
how do monocytes appear under a microscope?
have vacuoles
how do platelets appear under a microscope?
small
no nucleus
some granulation
what are platelets involved with?
haemostasis
formation of platelet plug
accelerating coagulation
inhibition of antithrombin III
in haematology what does the suffix philia or cytosis mean?
increase in number
in haematology what does the suffix penia mean?
decrease in number
what is thrombocytopenia?
reduction in number of platelets
what type of cells is the suffix philia used for?
granulocytes only
what is cytosis used for?
all other cells (apart from granulocytes)
in what species is the neutrophil the dominant cell type?
healthy cats, dogs, horses and camilids
in what species is lymphocyte the dominant cell type?
healthy cattle and rodents
what is WBC morphology like in rabbits and birds?
neutrophils are known as heterophils - have shiny pink granules like an eosinophil
in what species is the azurophil found?
reptiles and amphibions (monocyte with pinky/blue staining)
how do RBC appear in birds, reptiles, amphibians and fish?
nucleated
thrombocyte (platelet) is also nucleated - may look like a lymphocyte
counting is not performed by analysers due to presence of nucleus (cannot differentiate between RBC, thrombocyte or WBC)
what is used instead of a change in leukogram in horses, cattle and sheep?
haematology and measurement of serum acute phase proteins to detect inflammation
what can be diagnosed through blood smear?
thrombocytopenia regenerative vs non-regenerative anemia rouleux vs agglutination check machine WBC differentials/ manual differential WBC count normal and abnormal morphology
what can happen during automated counts of platelets in whole blood in EDTA?
done by analysers
commonly leads to artefactual (false) thrombocytopenia due to platelet clumping and macroplatelets
overlap in size between RBC and platelets often leads to false automated counts
under what circumstances should platelet numbers from blood in EDTA be checked by blood smear?
any animal with low automated counts
every animal with clinical signs of haemorrhage
what do platelet estimates on blood smears assume?
no platelet clumps on feather edge