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From bloodjournal.hematologylibrary.org by guest on August 9, 2012. For personal use only.

1957 12: 657-670

A Comparative Evaluation of the Sensitivity of the L.E. Cell Test Performed Simultaneously by Different Methods
EDMUND L. DUBOIS and VIVIAN FREEMAN

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From bloodjournal.hematologylibrary.org by guest on August 9, 2012. For personal use only.

A

Comparative L.E. Cell

Evaluation Test Performed by Different
I.1. DtBoIs descriptioms of this its
AND

the SensitivitySimultaneously Methods
\IVIAN FREEMAN

of

of

the

By

EmmIuNn classic

S

IXCE
mssethods

Hargras-es’
for the As each

of test

the has-c reports methods, of the

L.E. felt

in cell1948,

miumerous

performance technic In view merits of appeared

beemi

described that their

by method

sarious is-as to deterto perfornis blood antico-

authors.ui

proponent-sthe most mine the

sensitive. comparative a one

of the conflicting of the various different and and
AN!)

and the necessity it si-as decided

simultaneously draivn from agulants,

battery venipuncture

to

types study

L.E. tests on peripheral relative importance of on the L.E.

preservatives,

clotting,
MATERIALS

leukocyte
METHOI)S

trauma

phetiomenomu.

thissttndy the following four tecisusics were utilized after blood from a sirsgle venipunct.ure: 10 ml. was added to each of two test tubes containing 0.75 and 3.75 mg. of aqueous sodiunns heparin, another 10 cc. was permitted to clot, andfinally several drops were used to perform the Snapper-Nathan

Dtiring the taming 30 cc.

first phase of peripheral

of

ob-

ring
using

technic
threedescribed
drops of

below.t2
aqueous

The
sodium

amounts
heparin

of anticoagunlant
10 mg. per ml.

could
and 50

be approxinsated by
mg. per cc. obtained

through a 21 gauge needle. The preservative contained in the 50 mg. heparin was phenol 0.5% and in the 10 mg. concentration was chlorobutanoh 0.5%. Outlined below is the method by which we performed the test utilizingheparinsized blood. 1) The heparinized blood was allowed to remain at room temperature 30-60 nun. 2) The specimen was then centrifuged at approxinuately 1000 RPM or (104 X G) for

five
3)
4)

minutes,
The
One ml.

or until
supernatant
of the

a

clear

separation

of

the and

three

layers

was

obtained.

plasma
buffy coat

was
was

removed
taken

with

aWintrobe
placed or into (104 X G)

pipet
a Wintrobe for

and
tube. five

discarded.
minuutes, or

5) unntil 6)
7)

The Wintrobe a separation The supernatant

of

tube was centrifuged 1000 at RPM the three layers was obtained. plasma was again removed

with

a Wintrobe

pipet

and puushed

discarded.
ous glass to

buffy coat was carefully aspirated into the pipetandthen slides. In order to make a thin sections of the smear, the smearing slide was and then pulled backward over one-half width the of the blood film. This better study the cytology when the btnffy coat was very in leukocytes. rids 8) The preparations were stained with Wright’s stain in the usual manner. 9) The film was examined especially at the edges for at least 10 minutes mersion lens. In...