Page 702 - Clinical Hematology_ Theory _ Procedures ( PDFDrive )

686 PART 8 ■ Fundamentals of Hematological Analysis

SPECIAL HEMATOLOGY PROCEDURES (continued)

enter into the cell than leave. T is net in ux o water mol-
Normal Fresh Incubated
Range ecules eventually ruptures the cell membrane.
100 Hereditary
s Spherocytosis Te main actor in this proce ure is the shape o the eryth-
i
ys
l rocyte, which is epen ent on the volume, sur ace area, an
o 75
m
e unctional state o the erythrocytic membrane. A spherocytic
h

t erythrocyte ruptures much more quickly than normal eryth-
n 50
e
c
r rocytes or erythrocytes that have a large sur ace area per vol-
e
P
25 ume, such as target or sickle cells. T e ragility o erythrocytes
is increase when the rate o hemolysis is increase . I the rate
0
0.7 0.6 0.5 0.4 0.3 0.2 0.7 0.6 0.5 0.4 0.3 0.2 o hemolysis is ecrease , the erythrocytic ragility is consi -
NaCI (g/dL) ere to be ecrease . T e clinical value o the proce ure is in

FIGURE 32.11 Osmotic ragility (as mani este by percent hemo- i erentiating various types o anemias (Figs. 32.10 an 32.11).
lysis) o normal an here itary spherocytosis (HS) erythrocytes T is proce ure, in CLSI ormat, is provi e on this book’s

a er incubation in salt solutions o varying tonicity. In resh HS companion Web site at http://thepoint.lww.com/ urgeon6e.

erythrocytes, note the “tail” o cells with increase sensitivity as a
result o splenic con itioning (lef ). In the incubate re bloo cell SUCROSE HEMOLYSIS TEST

(RBC), note that the entire HS population o RBCs is more osmoti-

cally sensitive (right). (From Gla er BE, Naumovski L. Here itary Principle

re bloo cell isor ers. In: Rimoin DL, Connor JM, Pyeritz RE, Erythrocytes in PNH lyse when expose to serum solutions o

et al., e s. Principles and Practice of Medical Genetics. New York: low ionic strength containing complement. T is test emon-
Churchill Livingstone, 1996, with permission.) strates the sensitivity o erythrocytes to the protein, complement.

Normal erythrocytes un er similar circumstances o not lyse.

the membrane in equal amounts. In hypotonic solutions, T is proce ure, in CLSI ormat, is provi e on this book’s

erythrocytes will hemolyze because more water molecules companion Web site at http://thepoint.lww.com/ urgeon6e.

SPECIAL STAINS

ACID PHOSPHATASE IN LEUKOCYTES: than other leukocytes. Most o the aci phosphatase isoen-

CYTOCHEMICAL STAINING METHOD WITH AND zyme is inhibite by l-tartaric aci . T e cells o hairy cell

WITHOUT TARTRATE leukemia, Sézary syn rome, an some -cell acute lympho-

Principle blastic leukemias are tartrate resistant.

Peripheral bloo or bone marrow smears are f xe an incu- In the absence o tartaric aci , all leukocytes emonstrate

bate in a solution o naphthol AS-BI phosphoric aci an granular sites o enzyme activity. When incubate with tar-

ast garnet GBC salt. Naphthol AS-BI, release by enzymatic trate, an occasional granule may be observe in lympho-

hy rolysis, couples imme iately at aci pH with ast garnet cytes an some specialize macrophages (e.g., Gaucher

GBC to orm an insoluble maroon ye eposit at sites o cells an hairy cells). In bloo smears, a positive reaction is

activity. T is is the reaction that occurs at the cellular sites o enote by the presence o more than two cells with i use

aci phosphatase activity. an intense activity (i.e., more than 40 granules).

-
-
Naphthol AS BI phosphate aci phosphatase → naphthol Clinical Applications
AS BI T is proce ure istinguishes lymphocytes rom other types
-
o mononuclear cells. Because the cells o hairy cell leukemia

+
Naphthol AS-BI ast garnet GBC → insoluble are among the ew abnormal cell types that are tartrate resis-
maroon pigment tant, it is consi ere to be a signif cant marker in this isease.

T is proce ure in CLSI ormat is provi e on this book’s
Duplicate bloo or bone marrow smears are incubate in a

solution that also contains l-(+)tartrate–containing substrate. companion Web site at thepoint.lww.com/ urgeon6e.

Cells containing tartaric aci -sensitive aci phosphatase o ALKALINE PHOSPHATASE IN LEUKOCYTES:

not exhibit any ye eposits, whereas those mononuclear CYTOCHEMICAL STAINING METHOD

cells containing tartaric aci -resistant aci phosphatase are

not a ecte by such treatment. Principle

Most leukocytes exhibit a positive aci phosphatase reac- Leukocyte alkaline phosphatase (LAP) activity can be

tion to varying egrees. Lymphocytes isplay less activity increase , normal, or ecrease in a variety o con itions.