Blood smear evaluation and hematology
Dennis B. DeNicola, DVM, PhD, DACVP
Introduction
o Academic and commercial reference laboratories embrace the value
of the blood film
• Validating data
• Identifying key morphologic changes for all cell type
o In-clinic hematology analyzers are widely used throughout the globe
• Regional differences, but blood film evaluation is rarely performed
2
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Complete Blood Count: Data Evaluation
RBC mass
RBC morphology
Objective measure
of BM response
WBC mass
WBC relative
distribution
WBC absolute
distribution
PLT mass
PLT indices
3
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Introduction
o Academic and commercial reference laboratories embrace the value
of the blood film
• Validating data
• Identifying key morphologic changes for all cell type
o In-clinic hematology analyzers are widely used throughout the globe
• Regional differences, but blood film evaluation is rarely performed
o What can be done
• Continue education efforts both in veterinary colleges and with post
graduates
• Instruct usage and interpretation of various cytograms from analyzers
4
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Data, Cytograms and Blood Smear
Quantitative
Buffy coat.
Impedance
Laser
ProCyte
Dx
RBC
RBC
WBC
•
•
5
© 2016 IDEXX Laboratories, Inc. All rights reserved.
WBC
Help verify numbers in CBC
Help identify when a blood film review is essential
• Recognize abnormalities not identified with
numbers
• Recognize low frequency abnormal morphology
Blood film morphology – “Top Ten List”
Erythrogram
o Polychromasia
o Spherocytosis
o Agglutination
o Leptocytosis
o Acanthocytosis
o Hypochromasia
o Schistocytosis
o Metarubricytosis
o Heinz Body formation
o Miscellaneous inclusions
6
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Leukogram
o Normal leukocyte identification
• Five-part differential
o
o
o
o
o
Band neutrophils
Hyposegmented neutrophils
Toxic neutrophils
Reactive lymphocytes
Atypical leukocytes
Thrombogram
o Platelet number evaluation
o Enlarged platelets
Blood film morphology – “Top Ten List”
Erythrogram
o Polychromasia
o Spherocytosis
o Agglutination
o Leptocytosis
o Acanthocytosis
o Hypochromasia
o Schistocytosis
o Metarubricytosis
o Heinz Body formation
o Miscellaneous inclusions
7
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Leukogram
o Normal leukocyte identification
• Five-part differential
o
o
o
o
o
Band neutrophils
Hyposegmented neutrophils
Toxic neutrophils
Reactive lymphocytes
Atypical leukocytes
Thrombogram
o Platelet number evaluation
o Enlarged platelets
Erythron – “Top Ten List”
1.
8
Polychromasia – morphologic evidence of increased production in
response to a peripheral demand
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Classification of Anemia
Anemia
Regenerative
Anemia
Nonregenerative
Anemia
Absolute Reticulocyte count most objective method
of differentiating regenerative from non-regenerative
anemia
Polychromasia correlates with Reticulocyte count
9
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Reticulocytosis without anemia
No reticulocytosis
Reticulocytosis
87%
90.2 %
9.8 %
Retrospective review
- 285,169 Canine CBCs
- ProCyte Dx
10
© 2016 IDEXX Laboratories, Inc. All rights reserved.
13%
Retrospective review
- 7,421 Canine CBCs
- UK Reference lab
11
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Polychromasia
o Variation in color among cells
o Bluish color in erythrocytes (RNA)
o Cells generally larger than mature
RBCs
o Represent reticulocytes
o Few are normal in dogs and cats
o Good morphologic evidence of a bone
marrow response to a peripheral
demand
Problem in the field with use of rapid stains like Diff Quik
• Everything looks blue if not performed correctly
12
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Toby
o Signalment:
• 12 year old, intact male, Mixed breed dog
o History
• Current on all immunizations
• No previous significant illnesses
o Clinical Presentation / Physical Examination
•
•
•
•
13
Sudden onset depression and anorexia
Pale mucous membranes
Mild fever
Abdominal palpation – enlarged spleen
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Toby Erythrogram
14
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Absolute Reticulocyte Count vs RBC Indices
Normal MCV,
53.6 %
15.6%
8.3%
22.5%
MCHC
High MCV
Low MCHC
High MCV, Low
MCHC
6,752 (33%) Cases of
Canine Regenerative
Anemia
Retrospective review of laboratory submissions to
several national reference laboratories (200,196 CBCs)
15
© 2016 IDEXX Laboratories, Inc. All rights reserved.
o Retrospective study at UC Davis (2002 – 2008)
• 4,387 dog CBCs
33%
• 1,426 (32.5%) had regenerative anemia
8.3%
• 168 (11.8%) has macrocytosis and hypochromasia
• Based on reticulocyte count
o Blood film recommended (polychromasia) when a reticulocyte count
is not available
16
© 2016 IDEXX Laboratories, Inc. All rights reserved.
UK Reference lab – canine CBC results
180
163/851
(19%)
160
140
120
100
80
60
8.3%
25%
40
15%
10%
20
0
REG ANAEMIA
Non-anemic Samples =
All Anemic Samples =
Regenerative Anemic =
17
© 2016 IDEXX Laboratories, Inc. All rights reserved.
HIGH MCV
8278
851
163
LOW MCHC
HIGH MCV & LOW
MCHC
18
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Toby Erythrogram
Marked reticulocytosis
More consistent with hemolytic
versus blood loss
19
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Normal Erythrocyte Morphology
o Features similar in dogs, cats, horses
and ruminants
o All lack nuclei
o Stain reddish to reddish-orange
o Generally biconcave discoid-shaped
cells
o Major differences are in size and central
pallor
20
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Normal Erythrocyte Morphology
o Features similar in dogs, cats, horses
and ruminants
o All lack nuclei
o Stain reddish to reddish-orange
o Generally biconcave discoid-shaped
cells
o Major differences are in size and central
pallor
21
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blood film morphology – “Top Ten List”
Erythrogram
o Polychromasia
o Spherocytosis
o Agglutination
o Leptocytosis
o Acanthocytosis
o Hypochromasia
o Schistocytosis
o Metarubricytosis
o Heinz Body formation
o Miscellaneous inclusions
22
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Leukogram
o Normal leukocyte identification
• Five-part differential
o
o
o
o
o
Band neutrophils
Hyposegmented neutrophils
Toxic neutrophils
Reactive lymphocytes
Atypical leukocytes
Thrombogram
o Platelet number evaluation
o Enlarged platelets
Spherocytosis
o Appear smaller than normal mature
erythrocyte
o More dense staining than normal
mature erythrocyte
o No central zone of palor
o As name suggests, they have lost
their normal biconcave disc shape
and are “spherical”
o Supportive of extravascular immune
mediated destruction
• Partial phagocytosis
23
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Immune Mediated Hemolytic Anemia
Macrophage
24
© 2016 IDEXX Laboratories, Inc. All rights reserved.
FcR
RBC
Spherocytosis
25
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Toby
26
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blood film morphology – “Top Ten List”
Erythrogram
o Polychromasia
o Spherocytosis
o Agglutination
o Leptocytosis
o Acanthocytosis
o Hypochromasia
o Schistocytosis
o Metarubricytosis
o Heinz Body formation
o Miscellaneous inclusions
27
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Leukogram
o Normal leukocyte identification
• Five-part differential
o
o
o
o
o
Band neutrophils
Hyposegmented neutrophils
Toxic neutrophils
Reactive lymphocytes
Atypical leukocytes
Thrombogram
o Platelet number evaluation
o Enlarged platelets
Agglutination
28
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Agglutination
o Three dimensional clumps of
erythrocytes
• Unorganized in contrast to “stack of
coins” as with Rouleaux
o Relatively strong binding between
cells because of cross-linking of
antibodies on surface
o Supportive of an immune-mediated
mechanism
29
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Agglutination
Agglutination
30
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Rouleaux
Toby – body of the blood film
31
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Saline Agglutination Test
Relatively simple
Rarely performed
Over-interpretatoin of autoagglutination
0.9% Saline
Solution
Agglutination
32
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Rouleaux
33
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blood film morphology – “Top Ten List”
Erythrogram
o Polychromasia
o Spherocytosis
o Agglutination
o Leptocytosis
o Acanthocytosis
o Hypochromasia
o Schistocytosis
o Metarubricytosis
o Heinz Body formation
o Miscellaneous inclusions
34
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Leukogram
o Normal leukocyte identification
• Five-part differential
o
o
o
o
o
Band neutrophils
Hyposegmented neutrophils
Toxic neutrophils
Reactive lymphocytes
Atypical leukocytes
Thrombogram
o Platelet number evaluation
o Enlarged platelets
Target Cells - Leptocytosis
o “Floppy” cells with excess cell
membrane compared to normal
erythrocyte
o When associated with “regenerative”
blood picture
• Typical for immature erythrocytes
o When not associated with
“regenerative” blood picture
• Indicative of “lipid loading” potentially
associated with liver, splenic or
metabolic disorder
35
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Target Cells
36
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Kaos
o Signalment:
• 2 year old, intact female, Boxer dog
o History
• Current on all immunizations
• No previous significant illnesses
o Clinical Presentation / Physical Examination
• Slightly small for breed – smaller than littermates
• Slight decreased energy
• Occasionally has slightly loose feces
37
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Kaos - Erythrogram
38
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Lacey – Blood Film
39
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Kaos
o Clinical chemistry profile
• No significant abnormalities
o Urinalysis
• Ammonium biurate crystaluria
o Additional testing
• High fasting bile acids
• High fasting ammonia
o Diagnostic imaging
• Portosystemic shunt
40
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blood film morphology – “Top Ten List”
Erythrogram
o Polychromasia
o Spherocytosis
o Agglutination
o Leptocytosis
o Acanthocytosis
o Hypochromasia
o Schistocytosis
o Metarubricytosis
o Heinz Body formation
o Miscellaneous inclusions
41
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Leukogram
o Normal leukocyte identification
• Five-part differential
o
o
o
o
o
Band neutrophils
Hyposegmented neutrophils
Toxic neutrophils
Reactive lymphocytes
Atypical leukocytes
Thrombogram
o Platelet number evaluation
o Enlarged platelets
Acanthocytosis
o Specific type of poikilocyte
o Spherical shape with 2-10 surface
projections
o Projections are:
• Blunt
• Variably sized
• “Finger-like”
o Cholesterol:phospholipid ratio in RBC
membrane is increased
42
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Crenation
43
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Crenation
1 Hour
44
© 2016 IDEXX Laboratories, Inc. All rights reserved.
6 Hours
Acanthocytosis
o Specific type of poikilocyte
o Spherical shape with 2-10 surface
projections
o Projections are:
• Blunt
• Variably sized
• “Finger-like”
o Cholesterol:phospholipid ratio in RBC
membrane is increased
45
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Metabolic Disorders
o RBCs have phospholipid bilayer
Normal Red Blood Cell Membrane
Outer leaflet
Inner leaflet
46
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Metabolic Disorders
o RBCs have phospholipid bilayer
o Insertion in either the outer or inner leaflets can
cause shape changes in RBC
Outer membrane
cholesterol
Inner membrane
47
© 2016 IDEXX Laboratories, Inc. All rights reserved.
membrane lipids
Acanthocytosis
o Specific type of poikilocyte
o Spherical shape with 2-10 surface
projections
o Projections are:
• Blunt
• Variably sized
• “Finger-like”
o Cholesterol:phospholipid ratio in RBC
membrane is increased
o Supportive of potential liver, splenic or
metabolic disorder
48
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Sable – 10 yr, Fs, Mixed breed dog
o “Not feeling well” for about a week
• Not eating well
• Slight depression
o Physical examination
• Good body condition and hair coat
• Pink mucous membranes
• Tender abdomen on palpation
49
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Sable – Erythrogram
10 - 110
Normal RBC mass
Normal RBC indices
True Reticulocytosis??
50
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Sable – Erythrogram
10 - 110
Normal
Normal RBC mass
Normal RBC indices
Mild-to-moderate reticulocytosis
Clear evidence of reticulocytosis
51
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Sable – Blood film
52
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Sable – Blood film
53
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Sable – 10 yr, Fs, Mixed breed dog
o Case Outcome
• Diagnostic imaging of abdomen
• Radiographs revealed enlarged spleen and liver
• Ultrasound suggested blood-filled cavities
• Suspect splenic hemangiosarcoma
• Suspect hepatic metastases
• Owner elected surgical exploration
• Large splenic mass with evidence of hepatic metastases
• Owner elected euthanasia at surgery
• Final diagnosis - hemangiosarcoma
54
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blood film morphology – “Top Ten List”
Erythrogram
o Polychromasia
o Spherocytosis
o Agglutination
o Leptocytosis
o Acanthocytosis
o Hypochromasia
o Schistocytosis
o Metarubricytosis
o Heinz Body formation
o Miscellaneous inclusions
55
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Leukogram
o Normal leukocyte identification
• Five-part differential
o
o
o
o
o
Band neutrophils
Hyposegmented neutrophils
Toxic neutrophils
Reactive lymphocytes
Atypical leukocytes
Thrombogram
o Platelet number evaluation
o Enlarged platelets
Hypochromasia
o Pale staining cells compared to
normal
o Increased zone of central palor
o Supportive of decreased mean
cellular hemoglobin content
o In veterinary medicine primarily
suggests chronic blood loss
56
© 2016 IDEXX Laboratories, Inc. All rights reserved.
“Trevor” – 8yr, Intact male, Mixed breed dog
o “Not feeling well” for about a week
• Mild anorexia
• Slight decreased activity
• Referred to specialists because of possible anterior abdominal mass
identified with radiographs
o Physical examination
• Good body condition and hair coat
• Pink mucous membranes
57
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Trevor – Erythrogram
Mild decreased RBC mass
Normocytic
Normochromic
Mildly regenerative
58
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Trevor – Erythrogram
2
1
3
Things to note:
1. Reticulocytosis appears real (many purple dots)
2. Location of reticulocyte population lower than “normal”
3. Increased number of digitized events in small size range
59
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Trevor – Blood film
60
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Trevor – Erythrogram
Mild decreased RBC mass
Normocytic
Normochromic
Mildly regenerative
Low RET-He (Fe deficiency)
61
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Trevor – Case outcome
o Diagnostic imaging
• Mass in the wall of the stomach
o Surgical removal
• Solid mass with ulceration through the gastric mucosa
• Source of chronic blood loss
o Pathology
• Stomach mass
• Leiomyoma, neoplastic cells are within 1mm of the lateral surgical margin
• Moderate mucosal ulceration
o Follow-up
• Repeat blood work at suture material removal (2 weeks)
62
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Trevor – Case outcome
Trevor
3/10/2015
RET-He = 18.1 pg
Trevor
3/25/2015
RET-He = 19.6 pg
RET-He = x`22.1-29.5 pg
3/10/16
63
© 2016 IDEXX Laboratories, Inc. All rights reserved.
3/25/16
Maxi
Maxi
Patient
16 month old, intact male, Boxer
Presenting Complaints
• Monthly recheck while on prednisone therapy
• Maxi is normal according to his owners
History
• Immune-mediated meningitis diagnosed 9 months earlier
• Placed on prednisone
64
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Maxi – serial dot plots and RET-He
7/17/2015
HCT
7/17/2015
65
09/08/15MCV 11/10/15
Date
07/17/15
08/26/15
HCT (%)
(37.3 0 61.7)
41.1
11.2
25.4
25.4
41.6
MCV (fL)
(61.6 – 73.5)
65.9
40.7
44.7
52.2
60.3
RETIC (K/µL)
(10 – 110)
141.6
239.5
171.0
51.11
72.5
RET-He
(22.1 – 29.5)
25.1
13.0
17.5
19.6
26.7
© 2016 IDEXX Laboratories, Inc. All rights reserved.
12/22/15
Maxi – serial dot plots and RET-He
7/17/2015
VetConnect Plus
8/26/2015
HCT
Date
66
07/17/15
08/26/15
09/08/15
VetConnect Plus
11/10/15
12/22/15
MCV
HCT (%)
(37.3 0 61.7)
41.1
11.2
25.4
25.4
41.6
MCV (fL)
(61.6 – 73.5)
65.9
40.7
44.7
52.2
60.3
RETIC (K/µL)
(10 – 110)
141.6
239.5
171.0
51.11
72.5
RET-He
(22.1 – 29.5)
25.1
13.0
17.5
19.6
26.7
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Maxi – peripheral blood film
Relatively
normal
RBCs
Polychromatophils
equal to or smaller
than mature RBCs
67
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Small pale
RBCs
Maxi – serial dot plots and RET-He
7/17/2015
68
8/26/2015
9/8/2015
11/10/2015
12/22/2015
Date
07/17/15
08/26/15
09/08/15
11/10/15
12/22/15
HCT (%)
(37.3 0 61.7)
41.1
11.2
25.4
25.4
41.6
MCV (fL)
(61.6 – 73.5)
65.9
40.7
44.7
52.2
60.3
RETIC (K/µL)
(10 – 110)
141.6
239.5
171.0
51.11
72.5
RET-He
(22.1 – 29.5)
25.1
13.0
17.5
19.6
26.7
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blood film morphology – “Top Ten List”
Erythrogram
o Polychromasia
o Spherocytosis
o Agglutination
o Leptocytosis
o Acanthocytosis
o Hypochromasia
o Schistocytosis
o Metarubricytosis
o Heinz Body formation
o Miscellaneous inclusions
69
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Leukogram
o Normal leukocyte identification
• Five-part differential
o
o
o
o
o
Band neutrophils
Hyposegmented neutrophils
Toxic neutrophils
Reactive lymphocytes
Atypical leukocytes
Thrombogram
o Platelet number evaluation
o Enlarged platelets
Schistocytosis
o Irregularly shaped erythrocyte
fragments
o Indicate mechanical injury to
erythrocytes
o Damage often in small blood
vascular system
• “microangiopathy”
o Could prove helpful in
recognizing underlying
Disseminated Intravascular
Coagulopathy (DIC)
70
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Schistocytes
71
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Schistocytosis
o Irregularly shaped erythrocyte
fragments
o Indicate mechanical injury to
erythrocytes
o Damage often in small blood
vascular system
• “microangiopathy”
o Could prove helpful in
recognizing underlying
Disseminated Intravascular
Coagulopathy (DIC)
72
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Fleury – canine – 3/1/16 – severe inflammation
Fleury
Normal Canine
Mature red blood cells
Reticulocytes
Platelets
1. Note the clean separation of RBCs and Platelets as well as well
defined mature and immature RBC digitized events; these provide
confidence in the reported numbers
2. Note the increased number of events of “smaller” RBCs with Fleury
73
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Fleury – canine – 3/1/16 – severe inflammation
Fleury
The increased number of events should promote thorough microscopic evaluation
for microcytes, fragmented RBCs, and general poikilocytosis. In this case, there is
strong
forInc.
mechanical
74
© 2016 support
IDEXX Laboratories,
All rights reserved. injury to the RBC population.
Blood film morphology – “Top Ten List”
Erythrogram
o Polychromasia
o Spherocytosis
o Agglutination
o Leptocytosis
o Acanthocytosis
o Hypochromasia
o Schistocytosis
o Metarubricytosis
o Heinz Body formation
o Miscellaneous inclusions
75
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Leukogram
o Normal leukocyte identification
• Five-part differential
o
o
o
o
o
Band neutrophils
Hyposegmented neutrophils
Toxic neutrophils
Reactive lymphocytes
Atypical leukocytes
Thrombogram
o Platelet number evaluation
o Enlarged platelets
Metarubricytosis
o Metarubricytes are late stage
nucleated red blood cells
o Typically restricted from movement into
the blood
o Potentially present during:
Immune mediated
regenerative
anemia
• Marked erythroid hyperplasia in
“regenerative” marrow response
• Marrow stromal damage without
marked erythroid hyperplasia
• Acute lead toxicity
• Marrow infiltrative disease
• Septicemia
Acute Lead
Toxicity
76
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blood film morphology – “Top Ten List”
Erythrogram
o Polychromasia
o Spherocytosis
o Agglutination
o Leptocytosis
o Acanthocytosis
o Hypochromasia
o Schistocytosis
o Metarubricytosis
o Heinz Body formation
o Miscellaneous inclusions
77
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Leukogram
o Normal leukocyte identification
• Five-part differential
o
o
o
o
o
Band neutrophils
Hyposegmented neutrophils
Toxic neutrophils
Reactive lymphocytes
Atypical leukocytes
Thrombogram
o Platelet number evaluation
o Enlarged platelets
Heinz Body Formation
o Short blunt projections from surface of
cell
o Represent oxydized denatured
hemoglobin
• Onion toxicity - dog
• Acetominophen toxicity - cat
• Phenothiazide – horse
o Identification enhanced with New
Methylene Blue staining
78
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Heinz Bodies
79
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Heinz Bodies
80
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Tike - Physical Examination
Case study- Tike
Tike
Patient
7 year old, neutered male DSH
History
• 10 month history of PU/PD
• Referred as known diabetic
Insert
photo
81
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Presenting Complaints
• Lethargy and weight loss for last several weeks
Tike - Physical Examination
o
o
o
o
o
o
82
QAR, 5% dehydrated, mm pale pink, CRT 2 sec
TPR 103.1o F (39.5 o C),180 bpm,
BCS 2.5/5; mild generalized muscle atrophy
Activity score 3/10
Thoracic ausculation: unremarkable
Abdominal palpation: unremarkable
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Tike - Erythrogram
Remember – 5% dehydration
Probable minimal anemia
83
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Tike – Hemogram (day 4)
84
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Tike – Erythrogram (day 4)
85
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Tike – Erythrogram (day 4)
Tike
Note the increased density of digitized events in the region
where smaller erythrocytes would be found.
Microcytes
Schistocytes
86
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Tike – Blood Film
87
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Tike – Blood Film
Heinz Bodies
Heinz Bodies
88
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Tike – Blood Film – NMB Stain
Heinz Bodies
Aggregate Reticulocytes
89
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Heinz Bodies and “Keratocytes”
o Previous “blister cell”
o One or two projections that form from a ruptured
vesicle
o Usually seen with Heinz body anemias
Blister Cell
Keratocyte
90
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Heinz Bodies and “Keratocytes”
o Previous “blister cell”
o One or two projections that form from a ruptured
vesicle
o Usually seen with Heinz body anemias
Blister Cell
Keratocyte
Helmet cell
91
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blister Cell
92
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Keratinocyte
93
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Eccentrocyte Formation
o Fusion of opposed oxidized RBC membrane
• Potential accumulation of small portions of oxidized hemoglobin
between membranes (velcro)
o Leaves a pale area on one side of RBC
94
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Eccentrocyte Formation
o Fusion of opposed oxidized RBC membrane
• Potential accumulation of small portions of oxidized hemoglobin
between membranes (velcro)
o Leaves a pale area on one side of RBC
Side view
95
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Top view
Heinz Bodies and Eccentrocytes
96
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blood film morphology – “Top Ten List”
Erythrogram
o Polychromasia
o Spherocytosis
o Agglutination
o Leptocytosis
o Acanthocytosis
o Hypochromasia
o Schistocytosis
o Metarubricytosis
o Heinz Body formation
o Miscellaneous inclusions
97
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Leukogram
o Normal leukocyte identification
• Five-part differential
o
o
o
o
o
Band neutrophils
Hyposegmented neutrophils
Toxic neutrophils
Reactive lymphocytes
Atypical leukocytes
Thrombogram
o Platelet number evaluation
o Enlarged platelets
Miscellaneous Inclusions
Haemobartonella /
Mycoplasma
Distemper virus
inclusions
Babesia gibsoni
Basophilic stippling
98
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blood film morphology – “Top Ten List”
Erythrogram
o Polychromasia
o Spherocytosis
o Agglutination
o Leptocytosis
o Acanthocytosis
o Hypochromasia
o Schistocytosis
o Metarubricytosis
o Heinz Body formation
o Miscellaneous inclusions
99
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Leukogram
o Normal leukocyte identification
• Five-part differential
o
o
o
o
o
Band neutrophils
Hyposegmented neutrophils
Toxic neutrophils
Reactive lymphocytes
Atypical leukocytes
Thrombogram
o Platelet number evaluation
o Enlarged platelets
Five-part Differential
WBC
NEU
LYM
MONO
EOS
BASO
100
=
=
=
=
=
=
70.60
64.25
4.94
1.41
0.00
0.00
x10^9/L
x10^9/L
x10^9/L
x10^9/L
x10^9/L
x10^9/L
© 2016 IDEXX Laboratories, Inc. All rights reserved.
(
(
(
(
(
(
5.50 - 16.90
2.00 - 12.00
0.70 - 4.90
0.30 - 2.00
0.10 - 1.49
0.00 - 0.10
)
)
)
)
)
)
Common Leukogram Patterns
Moderate
Glucocorticoids
Epinephrine
Inflammation
(“Stress”)
(“Excitement”)
 - 
 - 
N-
Band Neutrophil
 - 
N
N
Lymphocyte
 - 

N-
Monocyte
N - 
N-
N
Eosinophil


N
Basophil
N-
N
N
Leukocyte Type
Mature
Neutrophil
101
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Inflammatory Leukogram Patterns
Leukocyte
Minimal
Mild
Moderate
Established
Overwhelming
Type
Inflammation
Inflammation
Inflammation
Inflammation
Inflammation
N
N-
 - 
 - 
 - 
N
N-
 - 
N-

Lymphocyte
N
N-
 - 
N-

Monocyte
N
N-
N - 
N-
N
Eosinophil
N
N-

N-

N-
N-
N-
N
Mature
Neutrophil
Band
Neutrophil
Basophil
N
© 2016 IDEXX Laboratories, Inc. All rights reserved.
102
Neutrophil Identification
o Most common leukocyte in most
animals
o 10-12 micrometers in diameter
o Nucleus divided into multiple lobes
o Condensed chromatin pattern
o Faint blue to light pink cytoplasm
o May see poorly distinct granules
103
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Neutrophil Morphology – Different Species
Canine
Feline
104
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Equine
Bovine
Neutrophil Function
105
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Lymphocyte Identification
o Second most common leukocyte in
most animals
o 10m in diameter
o Round in shape
o Round to oval, sometimes indented
nucleus
o Chromatin smooth areas intermixed
with more clumped chromatin
o Cytoplasm scant and blue
106
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Lymphocyte Morphology – Different Species
Canine
Feline
107
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Equine
Bovine
Equine
Bovine
Lymphocyte Function
108
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Monocyte Identification
o Largest leukocyte in peripheral blood
in health
o Round to angular in shape
o 15-20m in diameter
o Nucleus is variable in shape
o Chromatin is lacey
o Cytoplasm is blue-grey and often
contains vacuoles
o Looks similar in common domestic
species
109
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Monocyte Morphology – Different Species
Canine
Feline
110
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Equine
Bovine
Monocyte – Macrophage Function
111
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Monocytic and Macrophagic Inflammation
o Monocytes with differentiation to macrophages
• Erythrophage, Leukophage, Lipophage, etc.
Erythrophage
Lipophage
112
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Eosinophil Identification
o Absent or present in low numbers
o Slightly larger than neutrophil
o Nucleus similar to neutrophil
• Slightly less densely clumped
o Cytoplasm faint blue
o Multiple reddish to reddish-orange
granules
o Granules different shapes for most
species
Canine
Feline
Equine
113
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Eosinophilic Inflammation
1.
Parasitic infection
“Tissue Phase”
Paragonimus spp.
Canine Heartworm Disease - microfilaria
Equine verminous arteritis
114
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Eosinophilic Inflammation
1.
Parasitic infection
“Tissue Phase”
2.
Allergic or hypersensitivity reactions
Bronchoalveolar lavage fluid from
a dog with hypersensitivity
eosinophilic bronchitis
115
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Eosinophilic Inflammation
1.
Parasitic infection
“Tissue Phase”
2.
3.
Allergic or hypersensitivity reactions
“Other”

Association with various neoplastic processes
Mast cell tumor, malignant lymphoma, selected epithelial neoplasms, etc.

Various non-parasitic infections
Fungal agents

116
Foreign bodies
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Basophil Identification
o Larger than neutrophils
• Nucleus less segmented
• Chromatin less clumped
o Granules round and stain purple
o In cat, mature basophil granules do
not stain well
o High numbers of granules in horse
and cow
o Dog has only few granules
Canine
Feline
117
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Basophil Morphology – Different Species
Canine
Feline
118
Canine
Equine
Bovine
Feline
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Quantitative Buffy Coat Analysis
DNA
RNA
EOS
Plasma
RETIC
Top of
the float
Change
Bottom of
the float
PLT
119
© 2016 IDEXX Laboratories, Inc. All rights reserved.
L/M
GRANS
RBCs
Impedance technology
Heska
Difficult to accurately count
monocytes even with normal
samples
Abaxis
120
© 2016 IDEXX Laboratories, Inc. All rights reserved.
RNA content (Fluorescence)
Laser Flow Cytometry plus Optical Fluorescence
Granularity (Side Scatter)
121
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blood film morphology – “Top Ten List”
Erythrogram
o Polychromasia
o Spherocytosis
o Agglutination
o Leptocytosis
o Acanthocytosis
o Hypochromasia
o Schistocytosis
o Metarubricytosis
o Heinz Body formation
o Miscellaneous inclusions
122
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Leukogram
o Normal leukocyte identification
• Five-part differential
o
o
o
o
o
Band neutrophils
Hyposegmented neutrophils
Toxic neutrophils
Reactive lymphocytes
Atypical leukocytes
Thrombogram
o Platelet number evaluation
o Enlarged platelets
Common Leukogram Patterns
Moderate
Glucocorticoids
Epinephrine
Inflammation
(“Stress”)
(“Excitement”)
 - 
 - 
N-
Band Neutrophil
 - 
N
N
Lymphocyte
 - 

N-
Monocyte
N - 
N-
N
Eosinophil


N
Basophil
N-
N
N
Leukocyte Type
Mature
Neutrophil
123
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Identification of Band Neutrophils
o Definitions
• A neutrophil with a “horseshoe-shaped” nucleus with parallel-sided
nuclear membranes
• “Slight” nuclear indentations are acceptable
Band neutrophil
124
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Mature neutrophil
Identification of Band Neutrophils
o Definitions
• A neutrophil with a “horseshoe-shaped” nucleus with parallel-sided
nuclear membranes
• “Slight” nuclear indentations are acceptable
• A neutrophil where the thinnest portion of the nucleus is more than one
third the thickest portion of the nucleus
• Problem when nucleus is folded on itself
125
© 2016 IDEXX Laboratories, Inc. All rights reserved.
126
© 2016 IDEXX Laboratories, Inc. All rights reserved.
127
© 2016 IDEXX Laboratories, Inc. All rights reserved.
ASVCP/ACVP 2012
7
128
© 2016 IDEXX Laboratories, Inc. All rights reserved.
129
© 2016 IDEXX Laboratories, Inc. All rights reserved.
ASVCP/ACVP 2012
130
© 2016 IDEXX Laboratories, Inc. All rights reserved.
ASVCP/ACVP 2012
Blood film morphology – “Top Ten List”
Erythrogram
o Polychromasia
o Spherocytosis
o Agglutination
o Leptocytosis
o Acanthocytosis
o Hypochromasia
o Schistocytosis
o Metarubricytosis
o Heinz Body formation
o Miscellaneous inclusions
131
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Leukogram
o Normal leukocyte identification
• Five-part differential
o
o
o
o
o
Band neutrophils
Hyposegmented neutrophils
Toxic neutrophils
Reactive lymphocytes
Atypical leukocytes
Thrombogram
o Platelet number evaluation
o Enlarged platelets
Hyposegmented Neutrophil Identification
o Between a band and mature
segmented neutrophil
• Potentially 2 versus the normal 3-4
lobes
• Clearly hyposegmented
o Represents the stage of maturation
between band and segmented
neutrophil form
o Presence in significant numbers
supports presence of inflammation
132
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Granulopoiesis
133
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Granulopoiesis
Band Neutrophil
134
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Granulopoiesis
Band
???
???
???
Immature neutrophil forms
135
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Segmented
Neutrophil
Blood film morphology – “Top Ten List”
Erythrogram
o Polychromasia
o Spherocytosis
o Agglutination
o Leptocytosis
o Acanthocytosis
o Hypochromasia
o Schistocytosis
o Metarubricytosis
o Heinz Body formation
o Miscellaneous inclusions
136
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Leukogram
o Normal leukocyte identification
• Five-part differential
o
o
o
o
o
Band neutrophils
Hyposegmented neutrophils
Toxic neutrophils
Reactive lymphocytes
Atypical leukocytes
Thrombogram
o Platelet number evaluation
o Enlarged platelets
Toxic Neutrophil Identification
o Morphologic changes seen in
neutrophil forms with severe or
overwhelming inflammation
• Rapid maturation in response to
demand – skipping normal maturation
steps
• Potential direct effect of bacterial
endotoxins
o Potentially present in mature and
immature neutrophil forms
o Indicative of inflammatory disease
137
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Toxic Neutrophil Identification
1+
2+
3+
4+
138
Basophilia &/or Döhle bodies
Foamy cytoplasm
Marked basophilia & foamy cytoplasm
Nuclear degeneration
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Bands, Hyposegmentation and Toxicity
o
o
o
o
Nuclear shape
Nuclear chromatin – less condensed
Cell size – potentially slighly larger than mature neutrophil
Cytoplasmic staining – potentially slightly to moderately more blue
than mature neutrophil
o Cytoplasmic complexity – granule numbers, development, and
maturation
139
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Bands, Hyposegmentation and Toxicity
o
o
o
o
Nuclear shape
Nuclear chromatin – less condensed
Cell size – potentially slighly larger than mature neutrophil
Cytoplasmic staining – potentially slightly to moderately more blue
than mature neutrophil
o Cytoplasmic complexity – granule numbers, development, and
maturation
140
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blizzard
Case study- Blizzard
Blizzard
Patient
8 year old, intact male, English Springer Spaniel
Insert
photo
141
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Presenting Complaints
• Sudden onset decreased exercise tolerance
• Extremely weak
• Panting
• Fever
• Intermittent vomiting
Blizzard - Physical Examination
o
o
o
o
o
o
142
Quiet to depressed, ARH, mm pale, CRT 2 sec
TPR – T = 104.2 o F (40.1o C), P = 120 pbm, R = panting
BCS – 3 / 6
Activity score – 5 / 10
Abdominal palpation: WNL
Thoracic ausculation:
• diminished lung sounds on left
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blizzard - Leukogram
• Potential minimal leukocytosis
Qualified results
must be verified by
dot plot and/or blood
film evaluation
• Neutropenia, lymphocytosis,
monocytosis
• Suspect left shift
• Qualified (*) results
143
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blizzard - Leukogram
“Abnormal”
population of
cells
Location for
normal mature
neutrophils
Scan blood film!
144
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blizzard Blood Film
145
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blizzard – Blood Film
146
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blizzard - Leukogram
Immature and toxic neutrophils
147
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blizzard - Imaging
o Radiographs
• Pleural effusion
• Consolidated lung field
148
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blizzard - Additional Diagnostics
oIn-house fluid cytologic evaluation
oMicrobiologic evaluation – reference laboratory
149
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blizzard
150
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blizzard
151
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blizzard
152
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blizzard
Day 2
Day 4
Day 3
Day 1
153
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Day 8
Day 15
Blood film morphology – “Top Ten List”
Erythrogram
o Polychromasia
o Spherocytosis
o Agglutination
o Leptocytosis
o Acanthocytosis
o Hypochromasia
o Schistocytosis
o Metarubricytosis
o Heinz Body formation
o Miscellaneous inclusions
154
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Leukogram
o Normal leukocyte identification
• Five-part differential
o
o
o
o
o
Band neutrophils
Hyposegmented neutrophils
Toxic neutrophils
Reactive lymphocytes
Atypical leukocytes
Thrombogram
o Platelet number evaluation
o Enlarged platelets
Reactive Lymphocyte Identification
o Reactive Lymphocytes
• Small lymphocytes
• Increased cytoplasmic basophilia
• Increased quantity of cytoplasm
o Immunocytes (plasmacytoid)
• Small to large lymphocytes
• Moderate to abundant, deeply
basophilic cytoplasm
• Perinuclear clear zone
155
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Louie Lewis – canine – 3/2/2016
Louis
Normal Canine
11:09 AM
LYM
NEU
1. Lack of normal cluster for NEU
2. “Abnormal” cluster starting with NEU cluster and extending upward
3. LYM cluster appears shifted upward
156
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Louie Lewis – canine – 3/2/2016
Louis
11:09 AM
NEU
IVLS report: Bands – Suspect presence
The upward location of digitized events for the neutrophils is related to
increased amounts of cytoplasmic RNA, which is being detected on the
y-axis fluorescent scale. Note the dark blue staining cytoplasma
(increased RNA) and the poor nuclear segmentation (immature cells)
157
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Louie Lewis – canine – 3/2/2016
Louis
11:09 AM
LYM
Most lymphocytes on blood film have increased blue staining and/or
increased amounts of cytoplasm. Both indicate increased cytoplasmic
RNA explaining why they are located at a higher position on the
fluorescence y-axis.
158
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Louie Lewis – canine – 3/2/2016
11:09 AM
11:09 AM
Looking at a research tool to provide a higher resolution dot plot
confirms the suspected cluster evaluation on the IVLS dot plot
159
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blood film morphology – “Top Ten List”
Erythrogram
o Polychromasia
o Spherocytosis
o Agglutination
o Leptocytosis
o Acanthocytosis
o Hypochromasia
o Schistocytosis
o Metarubricytosis
o Heinz Body formation
o Miscellaneous inclusions
160
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Leukogram
o Normal leukocyte identification
• Five-part differential
o
o
o
o
o
Band neutrophils
Hyposegmented neutrophils
Toxic neutrophils
Reactive lymphocytes
Atypical leukocytes
Thrombogram
o Platelet number evaluation
o Enlarged platelets
“Atypical” Leukocyte Identification
o Large immature appearing nucleated
cells
o Typically mononuclear
o Potential morphologic features of
different cell lines
o Important to recognize
• Report to veterinarian
• Send to reference laboratory for
further characterization
• Likely to need special stains
o Potential underlying leukemia
161
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Louie Lewis – canine – 3/2/2016
Louis
11:09 AM
Normal Canine
Abnormal high
fluorescent cells
LYM
NEU
1.
2.
3.
4.
162
Lack of normal cluster for NEU
“Abnormal” cluster starting with NEU cluster and extending upward
LYM cluster appears shifted upward
“Abnormal” small cluster of high fluorescent cellular events
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Louie Lewis – canine – 3/2/2016
Louis
11:09 AM
Abnormal high
fluorescence cells
Significant numbers of high fluorescent cells with abundant nucleic acid
(RNA and DNA) content, which should prompt detailed blood film
review, especially at the feathered edge of the blood film. These
findings are highly supportive of a neoplastic lymphoid population.
163
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Abby – 6yr, Female, Labrador Retriever
o Original presentation
• Recurring epistaxis
• Physical examination
• Good body condition
• Mild splenomegaly
• Laboratory findings
• Marked thrombocytopenia (< 30 K/µL)
• SNAP 4Dx negative
o Being treated for immune-mediate thrombocytopenia of unidentified
etiology
164
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Abby – Leukogram
Moderate leukocytosis
Automated differential
Monocytosis / Eosinopenia
Suspect immature and/or toxic neutrophils (“BAND”)
Multiple qualified results (“*”)
Verify with dot plot and/or blood film review
165
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Abby – Leukogram
Questions to ask:
• Is there dot plot support for a left
shift?
• What is this abnormal cell
population?
166
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Abby – WBC Dot Plot
Abnormal population rich in nucleic acid content
Abnormal NEU
cluster – increasing
nucleic acid content
167
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Abby – blood film
168
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Abby – blood film
169
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Abby – serial WBC Dot Plots
170
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blood film morphology – “Top Ten List”
Erythrogram
o Polychromasia
o Spherocytosis
o Agglutination
o Leptocytosis
o Acanthocytosis
o Hypochromasia
o Schistocytosis
o Metarubricytosis
o Heinz Body formation
o Miscellaneous inclusions
171
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Leukogram
o Normal leukocyte identification
• Five-part differential
o
o
o
o
o
Band neutrophils
Hyposegmented neutrophils
Toxic neutrophils
Reactive lymphocytes
Atypical leukocytes
Thrombogram
o Platelet number evaluation
o Enlarged platelets
Platelet Number Evaluation
o Number of platelets per 100x oil
objective field of view
Normal platelet count
• Minimum: 8 – 10
• Maximum: 35 – 40
o Potential semiquantitation
• 20,000 x number of platelets seen per
100x objective field of view
Thrombocytopenia
172
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Blood film morphology – “Top Ten List”
Erythrogram
o Polychromasia
o Spherocytosis
o Agglutination
o Leptocytosis
o Acanthocytosis
o Hypochromasia
o Schistocytosis
o Metarubricytosis
o Heinz Body formation
o Miscellaneous inclusions
173
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Leukogram
o Normal leukocyte identification
• Five-part differential
o
o
o
o
o
Band neutrophils
Hyposegmented neutrophils
Toxic neutrophils
Reactive lymphocytes
Atypical leukocytes
Thrombogram
o Platelet number evaluation
o Enlarged platelets
Identification Of Enlarged Platelets
o Platelets larger than normal
Normal sized platelets
• Potential increased MPV from
hematology analyzer
o In the cat
• Usually equivocal finding
o In most other species
• Indicates marrow response to
peripheral demand for platelets
• Thrombocytopenia not required
• Inflammation and compensated
response by marrow
Enlarged platelets
174
© 2016 IDEXX Laboratories, Inc. All rights reserved.
Questions?
175
© 2016 IDEXX Laboratories, Inc. All rights reserved.