ENZYMATIC TECHNIQUES Flashcards
primary granules of granulocytic cells
MYELOPEROXIDASE
neutrophils
eosinophils
monocytes
lymphocyte (-)
MYELOPEROXIDASE
acute myeloid leukemia (AML) vs. acute lymphoblastic leukemia (ALL)
MYELOPEROXIDASE
lysosomes
all hematopoietic cells
T lymphocytes
ACID PHOSPHATASE (ACP) without Tartrate
resistance
preliminary identification of T ell subsets of acute lymphoblastic leukemia (ALL)
ACID PHOSPHATASE (ACP) without Tartrate
resistance
L-tartaric acid + staining substrate
TARTRATE-RESISTANT ACID PHOSPHATASE
hairy cells
TARTRATE-RESISTANT ACID PHOSPHATASE
hairy cell leukemia
TARTRATE-RESISTANT ACID PHOSPHATASE
diaminobenzidine (DAB) (benzidine substitute)
MYELOPEROXIDASE
brown reaction product
MYELOPEROXIDASE
copper salts followed by Gill’s modified Papanicolaou stain
MYELOPEROXIDASE
intense gray-black granules
MYELOPEROXIDASE
neutrophil and monocyte
MYELOPEROXIDASE
differentiates cells of lymphoid origin
from granulocytes and their precursors and monocytes
MYELOPEROXIDASE
Peripheral blood or bone marrow smears
ACID PHOSPHATASE (ACP) without Tartrate
resistance
TARTRATE-RESISTANT ACID PHOSPHATASE (TRAP)
ALKALINE PHOSPHATASE (ALP
ESTERASES
naphthol AS I phosphoric acid and fast garnet GBC salt.
ACID PHOSPHATASE (ACP) without Tartrate
resistance
enzymatic hydrolysis
ACID PHOSPHATASE (ACP) without Tartrate
resistance
Erastase
insoluble maroon dye
ACID PHOSPHATASE (ACP) without Tartrate
resistance
cellular sites
ACID PHOSPHATASE (ACP) without Tartrate
resistance
L- tartrate–containing substrate
TARTRATE-RESISTANT ACID PHOSPHATASE (TRAP)
Cells containing tartaric acid–sensitive acid phosphatase do not exhibit any dye deposits
TARTRATE-RESISTANT ACID PHOSPHATASE (TRAP)
mononuclear cells containing tartaric acid–resistant acid phosphatase are not affected by such treatment
TARTRATE-RESISTANT ACID PHOSPHATASE (TRAP)
leukocytes (+)
lymphocytes ()
TARTRATE-RESISTANT ACID PHOSPHATASE (TRAP)
tartrate resistant
hairy cell leukemia, Sézary syndrome, and some T-cell acute lymphoblastic leukemias
are
ACP IsoE 5
TARTRATE-RESISTANT ACID PHOSPHATASE (TRAP)
alkaline-dye solution
ALKALINE PHOSPHATASE (ALP
alkaline-dye solution
ALKALINE PHOSPHATASE (ALP)
naphthol AS-MX phosphate
ALKALINE PHOSPHATASE (ALP)
‘fast blue RR salt or fast violet B salt
ALKALINE PHOSPHATASE (ALP)
diazonium salt
ALKALINE PHOSPHATASE (ALP)
ESTERASES
insoluble, visible pigment
ALKALINE PHOSPHATASE (ALP)
Leukocyte alkaline phosphatase (LAP) activity
ALKALINE PHOSPHATASE (ALP)
leukemoid reactions and chronic granulocytic (myelogenous) leukemia
ALKALINE PHOSPHATASE (ALP)
touch preparations
ESTERASES
a Naphthol compound
ESTERASES
Naphthol AS-D chloroacetate
ESTERASES
alphanaphthyl acetate or alpha-naphthyl butyrate
stable diazonium salt
ESTERASES
highly colored deposits
ESTERASES
granulocytic vs. monocytic
ESTERASES
differentiation of leukemias
ESTERASES
• identify the cells of the granulocytic series. It does not stain lymphocytes and monocytes
Specific esterase (Naphthol AS-D chloroacetate)
• It can even be used to identify granulocytes in formalin fixed tissue
Specific esterase (Naphthol AS-D chloroacetate)
• In this procedure, blood or bone marrow smears are incubated with naphthol AS-D chloroacetate in the presence of a stable diazonium salt
Specific esterase (Naphthol AS-D chloroacetate)
• Naphthol compounds are coupled with diazonium salt, forming highly colored deposits at the sites of enzyme activity
Specific esterase (Naphthol AS-D chloroacetate)
• seen in monocytes (see Figure 5
Non-specific esterase (Alpha-naphthyl acetate esterase
• Both α-naphthyl butyrate or α-naphthyl acetate can be
Non-specific esterase (Alpha-naphthyl acetate esterase
The cells of granulocytic series are negative with this stain
Non-specific esterase (Alpha-naphthyl acetate esterase
The α -naphthyl butyrate stain is more specific, while the αnaphthyl acetate stain is
more sensitive
Non-specific esterase (Alpha-naphthyl acetate esterase
In a suspected myeloid leukemia, dual esterase stains can be used as they can
simultaneously classify the blasts as either myeloblasts or monoblasts
Non-specific esterase (Alpha-naphthyl acetate esterase
To differentiate positive reacting cells conclusively from monocytes, sodium fluoride is
incorporated with the incubation system
Non-specific esterase (Alpha-naphthyl acetate esterase
The monocyte enzyme is inactivated in the presence of this compound
Non-specific esterase (Alpha-naphthyl acetate esterase
Monocytic non-specific esterase is Fluoride sensitive.
Non-specific esterase (Alpha-naphthyl acetate esterase