3RD HEMA - Part 3 Flashcards
● Clinical syndrome resembling leukemia (Confused with CML)
● Not a disease, a description only
LEUKEMOID REACTION (LR)
● WBC ct.: >__ X 109/L (with neutrophilia and a marked left
shift [presence of immature neutrophilic forms])
(50) LEUKEMOID REACTION (LR)
● Excessive leukocytic response in the peripheral blood
LEUKEMOID REACTION (LR)
most frequently refers to neutrophils, but the increased
count may be due to an increase in other types of WBCs
LEUKEMOID REACTION (LR)
Generally used to distinguish LR from CML
LEUKOCYTE (NEUTROPHIL) ALKALINE PHOSPHATASE (LAP/NAP) TEST
Principle: ↑ LAP activity can be observed in neutrophils that have undergone normal growth.
LEUKOCYTE (NEUTROPHIL) ALKALINE PHOSPHATASE
(LAP/NAP) TEST
Principle: Hydrolysis of sodium alpha naphthyl phosphate by alkaline phosphatase produces a colored precipitate with a diazotized amine
KAPLOW’S METHOD
KAPLOW’S METHOD
: Fixative
Methanol and formalin
KAPLOW’S METHOD
Buffer:
Propanediole
KAPLOW’S METHOD
Substrate:
Sodium alpha naphthyl phosphate
mount in a mounting
solution like
Glycerol
KAPLOW’S METHOD
Initial stain:
Brentamine-fast garnet salt
KAPLOW’S METHOD
Counterstain:
Aqueous Mayer’s hematoxylin
No reddish brown to black precipitate
0
Moderately diffused reddish brown to black precipitate
2+
Slightly diffused reddish brown to black precipitate
1+
Very heavily diffused reddish brown to black precipitate
4+
Heavily diffused reddish brown to black precipitate
3+
Normal Kaplow’s (LAP) Score =
15 to 100
Decreased LAP score
Chronic myelogenous
leukemia (CML)
Normal or increased LAP score
Leukemoid reaction (LR)
Examples Of Cases With Increased Kaplow’s (Lap) Score
-Characteristics As Having
Leukemoid Reaction (Lr)
(4)
- Third trimester of pregnancy
- Polycythemia vera
- Infections
- Intoxication
Examples Of Cases With
Decreased Kaplow’s (Lap) Score
(4)
- CML
- Paroxysmal Nocturnal Hemoglobinuria
- Sideroblastic anemia
- Myelodysplastic syndrome
CML or LEUKEMOID REACTION
Leukocyte in the peripheral blood: Blasts/Promyelocytes
CML
CML or LEUKEMOID REACTION
Toxic granulation: Absent
CML
CML or LEUKEMOID REACTION
Leukocyte in the peripheral blood: Usually Myelocytes
LEUKEMOID REACTION
CML or LEUKEMOID REACTION
Toxic granulation: Present
LEUKEMOID REACTION
CML or LEUKEMOID REACTION
Eosinophils/Basophils: increased (↑)
CML
CML or LEUKEMOID REACTION
Eosinophils/Basophils: Decreased (↓)
LEUKEMOID REACTION
CML or LEUKEMOID REACTION
LAP: Decreased (↓)
CML
CML or LEUKEMOID REACTION
Splenomegaly: Usually prominent
CML
CML or LEUKEMOID REACTION
LAP: Increased (↑)
LEUKEMOID REACTION
CML or LEUKEMOID REACTION
Philadelphia chromosome (Ph1): Usually present
CML
CML or LEUKEMOID REACTION
Philadelphia chromosome (Ph1): Absent
LEUKEMOID REACTION
CML or LEUKEMOID REACTION
Splenomegaly: Mild (if present)
LEUKEMOID REACTION
CML or LEUKEMOID REACTION
Platelet count: Normal
LEUKEMOID REACTION
CML or LEUKEMOID REACTION
Platelet count: >600 or <50 X 109
/L
CML
Most common clinical application of flow cytometry: (2)
Diagnosis of Leukemias and Lymphomas
● Originally designed to evaluate physical properties of cells based on their ability to deflect light.
FLOW CYTOMETRY
- Measures physical antigenic, and functional properties of particles suspended in a fluid.
Flow cytometry
- Can describe antigen expression on many viable cells. Currently, ____ antigens can be detected at the same time on an individual cell. This is accomplished by the conjugation of monoclonal antibodies to a variety of fluorochromes that can be detected directly
17 antigens
Flow cytometry
Most significant discovery that led to improvement of flow cytometry and its subsequent widespread application in clinical practice:
Development of Monoclonal Antibodies
Although the term flow cytometry suggests the measurement of a cell, this procedure is successfully
applied to study other particles, including _____________, ______________, and __________.
Chromosomes,
Microorganisms, and
Proteins.
A flow cytometer is composed of
- Fluidics
- Light source
- Multiple detectors
- Computer
Laser =
Light Amplified by Stimulated Emission of Radiation
The basic unit of all radiation:
Photon
Commonly analyzed specimen in Flow Cytometry
(5)
● Bone marrow
● Peripheral blood
● Lymphoid tissue
● Body cavity fluids
● Solid tissue
● Must be processed within 24
to 48 hours from time of collection
For peripheral blood and bone marrow specimens
● Collected into a tube or
container with an anticoagulant (preferably heparin) and are transported to a flow cytometry laboratory at room temperature.
For peripheral blood and bone marrow specimens
This method, called
____________ __________, created a
central core of individually aligned cells surrounded by a
sheath fluid.
hydrodynamic focusing
The detector located directly in line with the illuminating laser beam measures _____________ scatter (FS or FSC), which is proportional to particle __________ or _______.
The detector located directly in line with the illuminating laser beam measures forward scatter (FS or FSC), which is proportional to particle volume or size.
A photodetector positioned to the side measures _____ scatter (SS or SSC), which reflects surface __________ and _______ structures such as granules and vacuoles.
A photodetector positioned to the side measures side scatter (SS or SSC), which reflects surface complexity and internal structures such as granules and vacuoles.
● CD 34
● CD117
● Terminal deoxynucleotidyl
transferase
Lineage
● CD 33
● CD 13
● CD 15
● CD 14
Granulocytic/
Monocytic
● CD 71
● Glycophorin A
Erythroid
● CD 41
● CD 42
● CD 61
Megakaryocytic
● CD 19
● CD 20
● CD 22
● κ Light chain
● Λ Light chain
B lymphocytes
● CD 2
● CD 3
● CD 4
● CD 5
● CD 7
● CD 8
T lymphocytes
__, __, and ___________ are displayed at the same time on
the instrument screen and registered by the computer system.
FS, SS, and fluorescence
Counterstain for __ to __ minutes
10 to 15 minutes
