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1251

37 xem —x SW666
undivided clones

April 21, 1955

56 (1) isolates, grow to 213 and summarily examined for motiles. Counts are
underestinates,. No tech losses
NG 22;2 Swarms 4 No motiles % less than9 22 10ormore 11

Maximum extimate: 40+. 5 clones were harvested for replaging of the intermediate

chains,
No, Est motiles harv. + nm. mot left behind Plate
B3 (18 20 4 ID) 3 oy: Lus T. Fee
B12. 28 22: 4 10) No lvsT leuk,
. I+fuaad, ri be but
Kh te co 20008 6 5 fat pet
fesihee Cl 30 37 Hvis, Coe 30'S
, Fy 10 10 34 swan + JS nm.
eat ” “ No-fracts,

) Estimates on clones with many chains are therefore moderately low. Some of the seros
ork may have had a motile but this was looked for. However, these drops were not searched
with a trap owing to shortness of time.

—

In addition, 4 drops had apparent swarms, but it was difficult to estimate incidence
of non-mbbile elements. Therefore these were blind-picked and plated immediately.
(picked to 10 ml, est. 60-70% recovery; plate .02 and .2 ml samples) (This will help
evaluate estimate of clone size as 2

Swarm @% mot, clone size logs Plate

c2 2 '0% wv . 0% Hw 43 1's

of, oe Ya '
out 70 40% 13 GF ae dee wal

pape" pegm 7 2 Ldormuristyd 06 «J
ee RNs ass

The data may be grouped as follows:

_ mots, clones Inchaba fue
iny Al 9 U1 B2 11 15 63 56 14.D 10 115 DB OF 12+ snl |
615-D1 0 A10 15 BSCE_DA AT 6
conf,in 1 A6 14 B4.7 10 12 C7 10 D2 9
tally bu 2 Bg C12 z
not pltg@’ 3 ‘A2 BS 14 C1 DZ Ss
4 A5 Bl C4 3
~~. m6 Ki2 ~ !
8 D6 DO 2
"10" A3 A4 D10 a
11-12 a8 B6 3
14 18 20 B12 B3 C9
20+ B12 Cl co = see above shh 3

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1253
Tetrazolium- formazan chains preliminary APR 3 4 teae

005%
P22. Prepare stained cultures. Add 1 ml overnight culture to 7 al broth + TZ .O9f$%
Incubate c. 3-4 hours. Also (A) add T% (1/200 .5%) to 1 ml culture directly.

Best method of preparation appears to be growth for short interval with TZ. Probably
only older nongrowing cells willstain.

‘Refr. to 1 P 23. Test isol. to agar, small liq. drops. Man traouble with agar is
confusion from dirt even under o1l. Probably better in fluid with a nonmotile culture.

1PM isol. 1) mot. W-2344 to Al

4PM 3 more to A2 >>:
4PM 6 stained W-2802 to small drops near situs C. These were termihally marked. ( b2}
_ Hunt for rare medial marked= 7,9 OA.

2 W-1177, (dividing) 9 is terminal, 10 is medial.

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—x SW 666; divided clones, TZ
1254

DATE: APR 25 1955 REF:

 

 

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Standard] sys tam now: SW-9L0 (FA 93;/H,~) @& tx SW 666.

  
  

0 .
A. Overnight SW666. Iml + 1 ml .01% TZ broth} Staion 9:00~-10330. Wash and add
FA 93 to|pellet. 11 AM- 1 PM. (Cf. B); sediment pellet for harvesting motile init.

A= ¢ells presbained; 2hours+ c
)

B. 1:1 +, FA93 9:30-LIAM. Add = vol.|TZ broth, Incuhats tillg stained (1PM). Sediment
to ah pellet, (31/2 hrs. + pha

 

 

10 B= cells poststained.

 

Found: y motile initials in each) but st no motile A were staindd (overstained?)
About of motile B were|hawelled; c. 50% of parent population.

 

Summary? | 28 isolates from B, 2 fromlA. 5 clones inviable. initially., jonly one Z

(granule) chain|died later. Zoqgckmn Z c s were) followed for 4 to 6 fissi
l clone gave a swarm (c. 50% motile) = 31B/ ee 7 neptles
?

Ucores autmonneg A P kaso ahr E. Thad oggpinaad
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2 P¥ stained W-1177 (c.
‘Plate out in mthocel broth on c.g.

t cells stain unipolar. Rat
terminal granule,

 

8:20 ~ 104K

 

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few definite

The bho bee Lawrmetteleyen Shp

edd oh To otf, te /2: Ox

probably from suppolar calls.

on . “s J2 95>
DATE: APR 28 1955 REF:
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