por #C a6 prepared at appropraste interven Cf
senihzed by autuchve. A volume Ot
ex,
ind
0.5 mb containing 50-75 pCig axkicd ¢ the
mm the centrefuge tube end mixed by gentle one
ai room temperature for 45 micutes!. Plasmg
excess Cr*! are separated from the tagged
tik
ceutnfugimg and washing thrice wah o Volum?
0.9 per cent sake soktion equal to thet of
.
decanted plasma, at room temperature The on
are resuspended in sufficics sabine
to Festone -
vohume to that of the original blood
santpite
either
used at ouce of refrigerated overnighto
4—5°C. The suspeimion is kept at
oun terery
dure for 45 minutes or longer befune it rawenius te
jecton Strict aseptic precautions are obverves
throughout.
Fig. }. Apparatus for delivering tagged cella. A =
calibrated 10 mi syringe, 5% inch needie; B =
ghaes tTesarverr (capacaics of upper and lower
chambers 35 mi and 15 ml, reopectively); C =
droposeble plastic tubing, /e inch deameter; D =
ghavs acheter.
of erythrocyte survival (Hughes Jones &
Mollison 1956).
The following report will describe and
assess a modification of Sterling & Gray's
method that we have used extensively (Noee et
mof et al. 1954; Wennesland, Brown, Hopper, Hodges, Guttentag, Scott, Tucker &
Measuring and delivering the dose of tagged cet
The subject's height and woight are recorded
and he rechnes ma comfortably wann room for a
beast 30 manmutes before the test. A wide-bore needle
connected to a small infusion apparatus (Fig |
ts placed én an antecubitad vein, The same madwelting
needic is used for dehvery of tagged cel!s and
walhdrawa!l of samples Enough 0.9 per cot satee
és pul wto the apparatus so that the bing
. * free, ;
of aie; the Jower chamber contains
2 wl,ant
the upper about 16 ml. Saline is allowed to ‘thy
slowly into the ver (o demonstrate the adequacy
of the venipuncture befbre the ocllsiare injected
‘ Converted
¥
Radioactryidy Research Cea
ained from the Oak Rider
fron Cr#Qly
Bradley 1959; Rapaport, Yamauchi, Green, ter
Nate Labor ony.
Brown & Hopper 1960; Brown, Hopper,
1 above is the one used
* The procedhire
1
Hodges, Bradley, Wennesland & Yamauchi). “at present, When we were collecting the data
we «in
forted a this paper, we were usiag 4 pias
METHOD
Cell tagging.
Vedi.
About 12 ml of ihe eubject’s
lagen cither 1" bebure of the momn-
tillation detector in which only $2 per cent of the
observed i
gamma disintegratrons of Cr! were
cans, 90 that 150-200 pC of Cr** were nented
ke
for lagen.
tarmal
"Tubes are fastened to a phorogeaph
revolving at J or pai.
oe ters he Wan Slyke 132)
i
Wes)
ert
subaatited
for
4 erg tice
the
needle
tise
ien
rome. Exactly Hot of the suspets
Oe
wt
Chanher of the me
delyveredh into the kevwer
raus (sce lig 1) ‘The syrume aanl
_
a tutalal Bla
agi ieraahet three ties with
r resersolr. ‘The
“_ ane takes fromm Ure uppe
ian ark washings TC alowed to) flow
-
pont the lower resersain ade the vem
8
be (elena
ay re followed by 4 sina aac bobrb
swcep ttn tarbuing dran
ol an which belps to
lhe
t Inde aes the
pene the aic Dubtde eaters the ver
period After Uns,
ni
masi
etre
de
the
of
beg ne
in
fe rate of infuse es celteced to the am
le C10 diagrnl
ecded to posure patclicy of the aecii
ed portions ofl the tapacesl
enue of fess}. Thre uous
e ol Meel wuspetewun is reserved for op
padouctivity.
|
Blood samples act taken AS nites or more
sher the begin of the misnog prermwl Ufsinelty,
two sanvples are taken al wi maceval of S annite~
wd the resuts averaged ‘To prevent dilution of
tee blood, wath sade, Che nelisgat as discontinued
i 30 sobonds before swnplin The first 2 3 nl
di bhad is discarded. AS ant samphe is then tikes
wd transferred to a test tube containing
elas
mahte mexture (Helker & Paul WOd4) Pupticate
Wazrube tubes are felled topped wath a small clioy
uf moucrad ait aad comtriiigel for A aia. al
(distawce frony Urea rag fee centre
luge comter = ES cm}. Dhey are reel te the top
@ the cell columnar Tae tot of the well nsed
vemple are deliver by
calebeated pipette dite a
fet tube (12 mm aterixe chaineter} ail cote
Veeoa well scitdlation comtec’. “Pwo 2 wl
1
.
The thiackwess of the butis coat is recorded
© Correclaon rs apqrlicd utiless st exeecds Foun
* With our Preset equipment, 45 per cout ot the
famma rays from the Cro! are alse reed as cont:
\ sallicrent Pumber of cunets (4080 of moter at
“Teed te
lant
the
1
of
the
statins ab
Cumeg to £ 16 per cou
fluctuation
ool
nesereal
cell
suspetseutr,
dilured
5) wath distilled water, are coumled: ur the some
hanes Coanetigs ol dkewal aid CoH suspenssar are
alternated fo nine the iaffaeace of possible
[hiktuations mi the scuseavity of the counter,
Caloulatrons
VC! Gal) -s
Ax 5SdxV
B
where A aml Hoace the averages of the observed
counts por seal per ol of the diluted cell suspenwom aial af thy boul specumens respectively, and
Vothbe volume em of cell suypensiun anyectedk
Cell yodume (Vile) is the product of VCrt! and
the average of the twe bematucnit readings of the
sanple, At this stage at 1s convenent to compare
the abverveadl Verbe with tbe predicted oclt volume
aba healthy sulgect of the same weight, height
aml sex, as established ly our studies wath thea
wetbou om 2H healthy men (Weonevkand ef al,
$959) goed TUE
Sampling and counliny
MOQ cpm
Portions
women (Frown af al). “Predicted
wormal’ values for Virbe can be fuwnd by a com
vermont grape anethod CWeusosland ef al 1959,
Wirtrobe 18Gb, Prown «f af) of by the following
—
4b t
4
A sobution Comaineng O.1—1.0 yg of N
was Caged
aerel test tube me whine Wout
for cou
eowhah bas bev calybeated)
an
WW
SS
2.8 mb of acid-citrate-dextrose solution i
ic i is clhawe frou,
ile tagged-cell suspene
nite
StaoNc Wade Seon
CUPL eS
Por
aen
Vide
bho x weil (he)
Heo
(cm)
- BW OSD. 190)
+
Vile Cink) = ? Sc beight (cm) 4
bar wontcn
Wa dx weight
(mil -- 86x beight
chy
O03 (ST).
134)
sdowikl be eter that trie cell volume is
shwhtly
sundher
thaw
Vide
as
cakukiled
above
Devatse the coll columis of the heanatocont tubes
fran
which the
sahe
is calculated conta 2-—S
ee
on
ing of the dest. Jin cases of severe
arma, nase:
blood is drawn to provide an
:
of cells. The bloud 6 injected throws
Wateay
cap of @ stembe §£5-mi Centrifuge tube he .
bebe
percent of teapped plana (Reeve 1952; Chaplin
&
Modliame
132.0
IMSS; Crrepedsca
A
Ebxugh,
Levee
&
Eanersou
Kaw 1059) aad the batty
Cont dsomecludel in the homer at peading. Our prethe Gone standiood Wome shed ef af 19599; Brown
eho) owete pooponed without curnrectiots fur trap
pal plasmna bectie segtle factur is utnfarmly
sepliadite CO lisplin & Modlin 1952; Ebaugh ef al.
55) Ali olevivel Vibe has been compared
woth othe “protec ponnal” for the sdbypect, a
switable cottoctien Lictot foe trapped plana may
ee
ik’! AUP bape BeeR Feb
——__
R. WENNESLAND ET AL.
oF
a
356