Romanowsky Stain /certified fixed orthodontic courses by Indian dental academy

RomanowskyRomanowsky
StainStain
Colour responses of blood
cells to
Romanowsky staining
INDIAN DENTAL ACADEMYINDIAN DENTAL ACADEMY
Leader in continuing Dental EducationLeader in continuing Dental Education
www.indiandentalacademy.comwww.indiandentalacademy.com

REFERENCESREFERENCES
 Shrish M. Kawathalkar; Essentials of haematology - 1Shrish M. Kawathalkar; Essentials of haematology - 1stst
editionedition
Jaypee brother medical publishers.Jaypee brother medical publishers.
 de Gruchi’s Clinical Haematology in Medical Practice- 5de Gruchi’s Clinical Haematology in Medical Practice- 5thth
editionedition
Blackwell Science Ltd.Blackwell Science Ltd.
 P. N. Marshall, S. A. Bentley, and S. M. Lewis ; StainingP. N. Marshall, S. A. Bentley, and S. M. Lewis ; Staining
properties and stability of a standardised Romanowsky stainproperties and stability of a standardised Romanowsky stain
Journal of Clinical Pathology, 1978, 31, 280-282Journal of Clinical Pathology, 1978, 31, 280-282

 P. N. Marshall, S. A. Bentley, and S. M. Lewis ; An evaluation ofP. N. Marshall, S. A. Bentley, and S. M. Lewis ; An evaluation of
some commercial Romanowsky Stains J. clin. Path., 1975, 28,some commercial Romanowsky Stains J. clin. Path., 1975, 28,
680-685.680-685.
 Text book Of Practical Haematology ; ninth edition, Dacie &Text book Of Practical Haematology ; ninth edition, Dacie &
Lewis.Lewis.

Colour responses of blood cells toColour responses of blood cells to
Romanowsky stainingRomanowsky staining
Cellular componentCellular component
NucleiNuclei
 ChromatinChromatin
 NucleoliNucleoli
CytoplasmCytoplasm
 ErythroblastErythroblast
 ReticulocyteReticulocyte
 ErythrocyteErythrocyte
ColourColour
 PurplePurple
 BlueBlue
 Dark blueDark blue
 BlueBlue
 Dark pinkDark pink

 PromyelocytePromyelocyte
 MetamyelocyteMetamyelocyte
 MyelocyteMyelocyte
 MonocyteMonocyte
 NeutrophilNeutrophil
 BasophilBasophil
 LymphocyteLymphocyte
GranulesGranules
 BasophilBasophil
 EosinophilEosinophil
 NeutrophilNeutrophil
 Toxic granulesToxic granules
 PlateletPlatelet
 BlueBlue
 PinkPink
 PinkPink
 Grey-blueGrey-blue
 Pink/purplePink/purple
 BlueBlue
 Blue/clearBlue/clear
 Purple / bluePurple / blue
 Red-orangeRed-orange
 PurplePurple
 Dark BlueDark Blue
 PurplePurple

Neutrophil Basophil Eosinophil
Monocyte Lymphocyte

ToxicToxic
GranulationGranulation
 SepsisSepsis
 InflammationInflammation
 Changes in granulocytesChanges in granulocytes
seen in blood film ofseen in blood film of
patients with inflammatorypatients with inflammatory
conditions.conditions.
 These are dark blue, coarseThese are dark blue, coarse
granules in neutrophils.granules in neutrophils.

Other inclusionsOther inclusions
 Howell-Jolly bodyHowell-Jolly body
 Dohle bodyDohle body
 PurplePurple
 Light blueLight blue

 Howell- jolly bodies are nuclearHowell- jolly bodies are nuclear
remnants in erythrocytes.remnants in erythrocytes.
 The nuclear remnants in normal redThe nuclear remnants in normal red
cells is removed by spleen.cells is removed by spleen.
 These bodies are seen inThese bodies are seen in
splenectomy, asplenia, hereditarysplenectomy, asplenia, hereditary
spherocytosis, megaloblasticspherocytosis, megaloblastic
anemia,abnormal erythropoisis.anemia,abnormal erythropoisis.

 Howell jolly bodies are round, smooth,Howell jolly bodies are round, smooth,
pyknotic purple bodies.pyknotic purple bodies.
 Size: 0.5-1 micron.Size: 0.5-1 micron.
 They are nuclear remnants or aggregatesThey are nuclear remnants or aggregates
of chromosomes that have been separatedof chromosomes that have been separated
from mitotic spindle and remain in RBC.from mitotic spindle and remain in RBC.

Dohle BodiesDohle Bodies
 11. Burn. Burn
 2. Infection2. Infection
 3. Physical trauma3. Physical trauma
 4. Neoplastic diseases4. Neoplastic diseases
 5. Chediak-higashi5. Chediak-higashi
Syndrome.Syndrome.
Döhle bodies are light blue-gray,
oval, basophilic, leukocyte inclusions located in
the cytoplasm of neutrophils.
They measure 1-3 µm in diameter. They are
thought to be remnants of the rough endoplasmic
reticulum.
They composed of agglutinated ribosomes.

MICROCYTEMICROCYTE
Iron deficiency anemia
Thalassemia
Lead poisoning

MACROCYTEMACROCYTE

BURR CELLBURR CELL
1. Chronic renal disease1. Chronic renal disease
2. Hyperlipidemia2. Hyperlipidemia

BURR CELLBURR CELL
 A/c echinocytes , red cells with abnormalA/c echinocytes , red cells with abnormal
cell membrane characterized by thornycell membrane characterized by thorny
projection.projection.
 It is reversible condition.It is reversible condition.
 Loss of intracorpuscular water due toLoss of intracorpuscular water due to
osmotic imbalance.osmotic imbalance.
 Red cell receptors bind to HDL whichRed cell receptors bind to HDL which
induces this shape in hyperlipidemia.induces this shape in hyperlipidemia.

TARGET CELLTARGET CELL
 1. Thalassemia1. Thalassemia
 2. Iron deficiency2. Iron deficiency
anaemiaanaemia
 3. Liver disease3. Liver disease
 4. Asplenia4. Asplenia

TARGET CELLTARGET CELL
 A/C codocyte.A/C codocyte.
 The appearance is due to decrease in HbThe appearance is due to decrease in Hb
content relative to surface area of redcontent relative to surface area of red
cell.cell.
 Decrease in cell volume without alteringDecrease in cell volume without altering
membrane area.membrane area.
 In blood vessel codocyte is bell shapeIn blood vessel codocyte is bell shape
and in smear appears target shaped.and in smear appears target shaped.

Sickle cell AnemiaSickle cell Anemia
1. Sickle Cell Anaemia1. Sickle Cell Anaemia

Sickle cell AnemiaSickle cell Anemia
Red cells containing HbSRed cells containing HbS
when subjected towhen subjected to
deoxygenated state leads todeoxygenated state leads to
polymerization (viscous gelpolymerization (viscous gel
state) of HbS molecule.state) of HbS molecule.
Upon oxygenationUpon oxygenation
depolymerization (fluiddepolymerization (fluid
state) of Hbs occurs leadingstate) of Hbs occurs leading
to normal shape of RBCs.to normal shape of RBCs.

Bite CellsBite Cells
 HaemolyticHaemolytic
disorder withdisorder with
GG6DP deficiency6DP deficiency

Bite CellsBite Cells
 Deficiency of G6DP leads to decreasedDeficiency of G6DP leads to decreased
availability of glutathione,which isavailability of glutathione,which is
important for removal of H2O2.important for removal of H2O2.
 Accumulation of H2O2 causesAccumulation of H2O2 causes
oxidation of Hb and ppt. of globinoxidation of Hb and ppt. of globin
which attaches to red cell membrane inwhich attaches to red cell membrane in
the form of inclusion bodies.(Heinzthe form of inclusion bodies.(Heinz
bodies)bodies)
 This red cell membrane isThis red cell membrane is
phagocytosed by splenic macrophages.phagocytosed by splenic macrophages.

Tear drop CellsTear drop Cells
 a/c dacroocyte.a/c dacroocyte.
 It is a type of poikilocyte.It is a type of poikilocyte.
 Seen in myelofibrosis.Seen in myelofibrosis.

 DISORDERS OFDISORDERS OF
WBCSWBCS

 LeukopeniaLeukopenia: WBCs count < 4,000/cumm.: WBCs count < 4,000/cumm.
 LeukocytosisLeukocytosis: 11,000-25,000/cumm: 11,000-25,000/cumm
 Aleukemia:Aleukemia:
 Blasts are not present in peripheral smear but are present in boneBlasts are not present in peripheral smear but are present in bone
marrow. Leukemic changes in bone marrow. ( WBC count ismarrow. Leukemic changes in bone marrow. ( WBC count is
subnormal)subnormal)
 Subleukemia:Subleukemia:
 Total leukocyte count is normal or low but blasts areTotal leukocyte count is normal or low but blasts are
demonstrable in peripheral blood.( WBC count is normal ordemonstrable in peripheral blood.( WBC count is normal or
subnormal)subnormal) www.indiandentalacademy.comwww.indiandentalacademy.com

 Leukaemoid reaction:Leukaemoid reaction:
 TLC is < 50,000/cumm.it can occur in infection,TLC is < 50,000/cumm.it can occur in infection,
inflammation and malignancies. presence of both mature andinflammation and malignancies. presence of both mature and
immature WBCs.immature WBCs.
 presence of toxic granules in neutrophils.presence of toxic granules in neutrophils.
 No splenomegaly, monocytosis, eosinophilia.No splenomegaly, monocytosis, eosinophilia.
 Acute leukemia:Acute leukemia:
 TLC is > 1 lakh/cumm.TLC is > 1 lakh/cumm.
 It is malignant clonal proliferation of immature or blast cellIt is malignant clonal proliferation of immature or blast cell
of haematopoitic origin.of haematopoitic origin.
 Short duration, fast growing , early appearance of c/f.Short duration, fast growing , early appearance of c/f.

 Subacute leukemia:Subacute leukemia:
 TLC is between 1-2 lakh/cumm.TLC is between 1-2 lakh/cumm.
 The severity and duration ranges between acute and chronic.The severity and duration ranges between acute and chronic.
 Chronic leukemia:Chronic leukemia:
 TLC >2 lakhs/cummTLC >2 lakhs/cumm
 It is malignant clonal proliferation of mature or differentiatedIt is malignant clonal proliferation of mature or differentiated
cells of haematopoitic origin.cells of haematopoitic origin.
 Long duration, slow growing , late appearance of c/f.Long duration, slow growing , late appearance of c/f.

Auer rods are clumpsAuer rods are clumps
of azurophilicof azurophilic
granular materialgranular material
present as elongatedpresent as elongated
structure instructure in
cytoplasm ofcytoplasm of
myeloblastsmyeloblasts..

Quantitative disorders of WBCsQuantitative disorders of WBCs

Neutrophilia :Neutrophilia :
 Acute haemorrhageAcute haemorrhage
 Acute infectionsAcute infections
 TraumaTrauma
 BurnsBurns
 IntoxicationIntoxication
 Metabolic disorders: acidosisMetabolic disorders: acidosis

Eosinophilia:Eosinophilia:
 Allergic reactionAllergic reaction::
urticaria,rhinitisurticaria,rhinitis
 FilariasisFilariasis
 TrichinosisTrichinosis
 Pulmonary disordersPulmonary disorders
 MyeloproliferativeMyeloproliferative
disordersdisorders
 HPV associatedHPV associated
carcinomascarcinomas

Monocytosis:Monocytosis:
 MalariaMalaria
 T.B.T.B.
 TyphoidTyphoid
 AMLAML
 SarcoidosisSarcoidosis
 Ulcerative colitisUlcerative colitis
 CarcinomasCarcinomas

Lymphocytosis:Lymphocytosis:
 Infectious mononucleosisInfectious mononucleosis
 MumpsMumps
 VaricellaVaricella
 ToxoplasmosisToxoplasmosis
 T.B.T.B.
 ALLALL
 CLLCLL
Atypical lymphocytes with indented nucleus, irregular cellular and
nuclear contours, dark basophilic lymphocyte with prominent nucleoli

Blood smear of acute lymphoblasticBlood smear of acute lymphoblastic
leukemialeukemia

Acute myeloid leukemiaAcute myeloid leukemia

Chronic lymphocytic leukemiaChronic lymphocytic leukemia

Chronic myeloid leukemiaChronic myeloid leukemia

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