See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/267228889

Blood Cell Morphology

Article · January 2013

CITATIONS READS

0 9,713

3 authors, including:

François Mullier Bernard Chatelain

CHU UCL Namur Université Catholique de Louvain - UCLouvain
345 PUBLICATIONS   5,145 CITATIONS    278 PUBLICATIONS   3,432 CITATIONS   

SEE PROFILE SEE PROFILE

Some of the authors of this publication are also working on these related projects:

perioperative management of DOACs View project

Pharmacovigilance and signal detection View project

All content following this page was uploaded by François Mullier on 01 November 2014.

The user has requested enhancement of the downloaded file.

Blood Cell Morphology

Claire LOOSEN, François MULLIER

Bernard CHATELAIN
BHS course, October 1th, 2011
 Blood cell morphology
• Validation of the hematology counter
parameters (PLT count, MCV…)
• Lack of sensitivity and specificity of the
hematology counter alarms (lymphoblasts,
Immature granulocytes…)
• Answer to questions coming from
– Biological results
– Clinical situation
 Material and Methods
• K2EDTA Sample
• Smear
– Wedge smear
– Spun smear
– Automated device
– Buffy coat (rare cell detection, diff if low WBC
count…)
• Staining
• Observation (classical, image capture and analysis)
• Interpretation
BLOOD SMEAR (Wedge)
Hemaprep
Diff spin

• Higher
monocyte
percentage
• Lack of
Smudge cells
in CLL
 May-Grünwald STAINING Check the pH
of the
Reagents Sorensen
Sorensen Phosphate Buffer pH 6.8 : Na2HPO4.2H2O M/15 (2.56 g/l) and KH2PO4 buffer!
M/15 (6.63 g/l)
Buffered water : dilution 1/20e of Sorensen Phosphate Buffer in distilled water
May-Grünwald solution
Don’t adjust
Giemsa solution
the pH of the
buffered
Staining water!
Methanol 10 minutes
Pure May-Grünwald solution 5 minutes
May-Grünwald (50/50 in buffered water buffer pH 6.8) 2-3 minutes
Giemsa (1/10 in buffered water pH 6.8) 15 minutes
Rinse with running water
Buffered water pH 6.8 30 seconds
Let dry smears upright

Guidelines for blood smear preparation and staining procedure for setting up an external quality assessment scheme for
blood smear interpretation. Part I: control material (EQALM), J-L Vives Corrons, S. Albarède, G. Flandrin, S. Heller, K.
Hovarth, B. Houwen, G. Nordin, E. Sarkani, M. Skitek, M. Van Blerk, J-C. Libeer, Clin Chem Lab Med, 2004, 42(8):922-
926

EQALM : External Quality Assurance Programmes in Laboratory Medecine

 Microscope
• High resolution objective (highest N.A.) 60X
1.4 N.A. oil i. planapochromatic
• Kohler setting with condenser whose N.A. is
matched with the objective N.A.
• Condenser diaphragm adjustment: full open if
N.A. close to 1.4.
 Red Cell morphology

Dacryocyte Schistocyte Codocyte

Drepanocyte Spherocyte Jolly Bodies

Codocytes
Dacryocyte (Teardrop cells)
Schistocyte definition

JF LESESVE (Hématologie)
SPHEROCYTE
Plasmodium falciparum
Platelet Morphology
Platelet agglutination and aggregation
• Variable ensitivity of the automated
hematology analyser
Platelet satellitism
• Lack of detection by Automated hematology
analyser
 Grey platelet syndrome
Lack of detection by automated hematology analyser

Patient from Jolimont: C.Chevalier, G.Detry

 MYH9-related disease

Giant platelets Döhle bodies

Platelet Lack of detection by automated

distribution (MPV) hematology analyser
and count 19
White Blood Cell morphology
 Lymphocyte morphology (t < 4h)

Cytoplasmic Cytoplasm Nucleus

membrane
• N/C ratio • smudge cells (lymphocyte
• smooth or irregular count + good prognostic factor in CLL
• Basophilic cyt, golgi but not specific of CLL)
• mobility, fragility, pattern
• Granules, inclusions • shape
• chromatin structure
• nucleolus (i)
 Lymphocytic population

Benign ? Neoplastic

Polymorphism Monomorphism

lymphocyte
immunophenotype
 Reactive Lymphocytes
(infectious mononucleosis)

Immunophenotype :
CD3+/ CD8+/ CD38+/ HLA-DR+ ou CD19+ activés
Binucleated polyclonal B lymphocytosis
Chronic Lymphocytic Leukaemia
B Prolymphocytic Leukaemia
Mantle cell lymphoma
Mantle cell lymphoma
Follicular lymphoma
Lymphocyte mobility
Hairy-cell leukaemia (1)
Hairy-cell leukaemia (2)
Splenic Lymphoma with villous lymphocytes
Lymphoplasmocytic lymphoma
 Marginal zone lymphoma

1. Villous lymphocytes
2. Plasmocytoid
3. Cleaved cells
4. Clumped chromatin
5. Nucleoli
6. …

!!! pleiomorphic
 Lymphoplasmocytic Lymphoma

• Red granules without peroxydase activity

Cytoplasmic inclusions in B lymphomas

S. Daliphard et al., Hématologie 2006; 12(1): 77-79

Acute Lymphoblastic Leukaemia
 Keep in mind…
• Granules may also be present in B lineage
• Neoplastic population may some time be present in low
percentage
• Smudge cells are not restricted to CLL
• Consider always the DD with Acute Leukaemia

MORPHO

CMF CLINIQUE
 EM (Blasts)

ALL AML
Phase Contrast Microscopy: blasts
Belgian EQA march 2010
 Belgian EQA march 2010

PEROXIDASE: negative

ALL
Sang LAM M3
Quality Assessment and Education
 Belgian EQAS
2004 November ALL
virtual microscopy
2005 March Normal, B Prolymphocytic Leukaemia
2005 June Normal, Hereditary Spherocytosis
2005 November Normal, Secondary leukaemia
2006 March Normal, B12 deficiency
2006 June Normal, Drepanocytosis, May Hegglin (didactic)
2006 November Myeloma, Chediak-Higashi (didactic)
2007 March Normal, CML
2007 June Normal, Reactive Lymphocytosis
2007 November Normal, Hairy Cell Leukemia
2008 March Normal, TTP
2008 May B12 deficiency, Secondary leukaemia (didactic) (bone marrow aspirate)
2008 June Secondary ALL, BiN Polyclonal Hyperlymphocytosis (didactic)
2008 November T Prolymphocytic leukaemia, MDS Ider20Q (didactic)
2009 March Hairy Cell Leukaemia, t(8;21) AML (didactic)
2009 June Normal, T cell large Granular Lymphocytic leukaemia
2009 November Normal, Leukemoid reaction
2009 November Myeloma (bone marrow aspirate)
2010 March CLL, Mucopolysaccharidosis Maroteaux-Lamy syndrome (didactic)
 Belgian EQAS
virtual microscopy
2010 June Normal, Plasmodium Falciparum, TTP
2010 November Normal, Sezary Lymphoma
2010 November Mantle Cell Lymphoma
2011 March Normal, Acute Lymphoblastic Leukemia
2011 June Normal, Acute Myeloblastic Leukemia
www.e-hematimage.eu
 References:
• Repères en diagnostic de laboratoire, X.
BOSSUYT, J.-M. BOEYNAEMS Ed: Garant ISBN
90-441-1022-5
• Wegwijzer in Laboratorium-diagnose, X.
BOSSUYT, J.-M. BOEYNAEMS Ed: Garant ISBN
90-441-1021-7
• Blood Cells A Practical Guide, Barbara J.
BAIN, ISBN 1-4051-4265-0
• Bone Marrow Pathology, Barbara J. BAIN,
David M. CLARK, Bridget S. WILKINS, ISBN 978-
1-4051-6825-0
View publication stats