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Wie FALOC (2/467 HneAid-tacts thoes drof hea To cmd etontr> V7 dese and W967, We ye lof ful. he 2 aepetilcn p FOB G67 : ° (unt gots coc Hy OA GC gut thes lea COC. o Ava | rf 767 *. 4 pmagtps /22 21K | Cakrgwr aly bo \ acl aeapnded cl ow auth oc aungpoed 22711) IGE [ie] Fr 2-3 are ches bin 9 |ucetinan, alls comple 2), EY 494 medtle chase fs « eal om gaat a toatl stage. (fiierrn ?) PS: teeT 1-3 fn Aly wiranee ond x Fito. (yuri Herarerer / E/~3 prt be galactone poreclot SEV os Sel inl a peuorer error bes ben rade, et dart EF 3 wer pryerslor FIAT Gal [flab Conbel Gallx be SW 957 ? whncze > [**5') Teal om ENA aGR. - Thee en a smal pratt oY theme wy W299 =lae® SR Dog dt- WLI WBE pre an egverylecs Ue . come dpa! Matting 9 Clea caigh ats Mfr fom fs ease mya abe pe tperesdinny Aumnrny cqrig lest for. Ror apohareg. a bect8! —— =—— cee ce > ae V/ FAb0C = 22/967 ad EG th 6. pate, , e FALL? BOC XE/-3 Gar NO FUneA. i catid —K 67 awe 7¢S. a A SWI? x Sebbee. M xed Phi port , sesmterees soa Noss famaductetotpose ) ( PAr% J) ~ 6 MA t chet. — Yer. ~%9b7 , 666 te curtth | Grune Fassendnon’ of hey W465. UJ 1404 Tw comednae G bp” c be tere) nuladnes hav elie twterobed) WTF and WU-/6 5% A. A Hid canetincled taal >> bac, bes rucerthin etd forfinco- ductins exbinbeage 1 bp, ( [2 eg cite thle chad Prat werteinnca, ys airtArgrlty , A. O prisgole WY oo Agaghed. Firef hte le ba publentally nl pe Len N Yeoh Dard 2 B00 Por sucatot thd | np wideteling ’ > ON wile aRerescgeeyl M; Ho tarot Cs A Otter TD ke pp. lec bE 2H sisi sled lta lp OL te nba Crrbedie tir WPS, IO Ardged Ui pancdduclin Fiqh af Cheba Hac fre Healt © boleh DCF aloe loch Jr other Aras C he’. P| ILE cackt] 3 [ bac bl) fn cilon. Taal by Porn? tone Hare wnt tryatillot hector ‘ wth EL fi Gaur an traced lp” auth td PA Bo EE [isd Apel 1974. Drdtin up Wormitdle Merrett Rha. Firstar 2% howe} 6/0. Ape ide Lhrbewer ~ 3:35. opis dirk af bach mee be ! Yo tadelad bw. fe 2 Q a amie, y ae othr "soe “atl Al OO as So) ARDB— > Sue Hirde ctatinnyelee GOGO — ed @ 6 (2 ES QO @ 6) & FB: @ ed & 14 O H3 &@/) & | H4 F | 63 62) Pr | F2@ Wo - ta sling cella of A? [1267 3 - FS), VK (an stile. spc Muntlowr © rn byt le oboe EX EY waar tor thy Alf vos berth Mod dette ihe ly pedisect De se ——x peed i c foe. __ y bl 4007. | a oo 221 578. Le A -FY. [27] Bh. Bo" PIP Threat anil colle. Hr iterate Q mre $5 jf dod | IZ panntde aH 10 *. bo WO 578 SOT; IS IT. _ 6% 20 bo ~« $34 © / s3/f © : sBdyY ft Pahebt Mibu eon siurtcatenate or BAMLTIAWKE sta C. i “rl | i bua, © Yar | Mota lk 4 “4 rey. dele mitcle uth, ae Li tajen a Mik z _* I ton Shpall ur yer, ' nl agp W- YB = C reed Manas fr ep elgcf) Gti e W209 a Are fat) esr, fat TA ty ) 9 psi fo fi = Ly, yy foc aon lg eal eb ieee fs, NE ILdDRL An becth (32°) . yo yt ofall. Due. {fr | tort f jp mdeEX [120A 37% 29°, /9 peered. 62076: ve rene j verve ee # Oy ot bunk mare ga wo att altnne “mM = = SIG. ql]: 1 cathe pine agp 10° bine | Retarcaden 30° Rnd 2° preuner te tho Aawme , BPI. Pid: eo spoken MK Bibeln “19 (Bh tnt fh, tacginhe, TOO [se 1 tarps om gehn- vetiby oe iy “iss Pid cig He > { re PIs both. Aheee 30 MITA. ZA Dd 40: 50: + 2YDE, W2L¥5D @ Fada sear >176-Gb- 108 (wun) Chat an 5 4,10 wW 1377, 395-97 x wWlt7) nfia[s B/6 — wyneh waz Wd th/rs Karis . . 13% pvaestd, "77 fe % (, Mi, Glas ‘Pee oi 10 20 30 40 > (re decors }) 4 D iy Ah t/O-/>), fed of aes ooh | ae. jr DATE: . REF: q710-( hex Ink ratle dba oro ho Nek phage tha.) fe, bal Fin 57 /1unk Sct hl (dd dil I |’ bist. ) ad Fdetgde db Aper 2 N SS Motos — Sys | {4 o> 1S" 2°5 /® (#4 TF Hibllethey hago iq “ee t NO x Sag 4 1955 Malis Peurihat | by Fe 7,/0 , » @.fO | Avt ia bev lhe 1%p ages ¢ Archangel (hdEA¥7 + Ing saced) PA 3? —m SW6GE JUS spose tome alge LTaNe raceme DATE: JAN 19 1955 REF: Deh aleep. See (Ap). A.tluk subbb+ fh wh FAT |. Feat Meryefils (Ler bye) | : by P- ol ui . ty patty “Zan. Poahummarey spy} , I No Cel cof cart ry ’ taf ,000] " T+8 jord dhe | 7g rrawol JAN 20 20 : pith Jo-3 lds, pfbebe a | Z (>Ss) vs nia] attt ) xjo~% ( lek lak beth | £47) x ION It [ek bee, ten". swb& 40 $y 22 Aener I. a ffee THA goede pat ohana fromm cherrse Peach falda Pe Syne OA Me ore fed obs tcae anu of Txaks : wdaspaten fucls b gvrarree ? hater rate i cabratis O guemns > O pt ehCt rh pitino w Tapble; fests wa (hee tubes? > ? en lly oe tn i i ig B7~XSWb66. (adte| ell, path, thine. Lintrose | [ Sebreaulls) iT QNN BO. 1955 JAN 1 1955 benreetion with bene rasa Fla. ~ Lented. jrecbsety ees 2. Fm free i H Leb Clr aac SLIZ =Swi0ye pur At , S128: MW/07r = hulls SOY 3616 pr ® 50 14+ Schuutge 0 $Wsv3 - Atty * SW9LE (Mart 0 6) / ” callin pear ; Mad vot yemap heres a a“ a ee ue "0 ee + #3 ry OF : } . -), . aoc i D i) wyame, | ome 05 (IT) 18,9 ; woh a ; Pale pred rye? aa) Angel, Hilo alrrocert, Q, aN Ahigu4 . cal od \ REF: [21 50 p03 |, Ra, Delyrry andl wer Fao4s Aaks : he 7e- /255 DATE: pel REF: (NMAC. ea IF SWE pute patel dy jponr. é rT fp "p> >. oe LEMS ot aerial aurie boel , | BAL FAB? 0. fing I poke ; ecslree c. J 40 20 NN 30 LS 50 5 Vos) Ife ol A) tharnan rata, Un tgictele len 07 bin, REO WI Mledb the co etn Tid fltator | Pacrred kal dora dr Mew pits w0 foto (ove ed et y— FA =42T:F9S ' Gsm, GOCE x— Fain - BET: 3S JAN 2 4 1955 [hanes Aatn a Ail JAN 2 4 1955 ee < Ccpecatin Mat T en gas La Swhll Aaptond ? ~ KX G67 Jere fod mean Calor Ad food beqoy ler Jura , (ra faker _xsw/09F JOT: (05. fe 194 Puls g Uvprialeal FAIL UV, ; 24T : O Stu aire, GT : dSeueune } he ott slab Cvat FAIL F437? (510 543? Wao bbb tl ! gay, FhUI—x* Swted YT 2 10$ rbot ~ PAV-¥ SwSBY 40T:VS Colin ll E muall | JAN 25 1955 Bas pst LC an pneid or 4. ) auspherin — (Ogun f fawsdutin (Lop breyitng) Bibi 9 Phoxs Me lade ae @) may) 7 () Plu. wforgree —K 6 halls OAaypeging- pt (bE) . puspinded A) ) (‘see 6 2) | bse ft, —x lenei ( 29. .) pewliftac a. : Vubeumndat Bgucke Gad, Sine tas os pully Tech aXeagh oars. on tarrlanies yyy Ti) Aebes hes Man rounimle fr Us woe? foe tonguuson & pkige ole ude. 999 PAIK pet 3P7:35$ BT A SS. PKS clam =——K SHUI )? x10 rhatt! / Sik Parle) quae ca 2D papally ! . S54) Lf a . + TD x20 Jot 10" pape pon phot | jo’? fo’? PMS claws - ld eA = SKso® + ! qual yl “WH FR. caput F423 Suwbb S22 oats) Che OP ~ devs. fy ar rang Stiri « JAN 2 @ 1955 SS 534 —, (clase ay10? TJecbl ( Yx16 “18 feo bf Joy oy | yup 7 pov. gull eregell be rato fe 5 tx V/ ? foe a eth Ce rere sx3 EC Maricneetreke tabis i, yy r nv Ce . ) ( cael tacky CAN ‘JAN 2 6 1955 | S Corminds . . Si SY I X— TNQD— dg. ToS. elf” gute 80-70 oh, wi 15H, 3" |b Kettle bdo puto tan 60 dr drdgpe loon ty Saplion afer TOK oh wnt igke! — Lal? 14 be sGIA clays lap. “Dat ama. Sa. caus nylo ats. lala Yudla . Ins. 190 IT< 10 T>/0 | Pitre [5 “ "E whe) Ye Bells 1st : Alar | 10 2,0 bil del 2a) G3 _ 30:3 44:3 Ta ect Eth at Ithavrd putin DATE: pe 27 ae | REF: 3 4 5 6 7 tac | A alate tite fag #2). cet fae o hn 42 i/ [0 Aff? brdshr- £Y +0 af pf oP )'Brr see . gan otis! =~ kyso hk po -— ie 2 | 2 ys egeato $e37T i as COU pp. 7 2° B poke of f’ DL Yauerr yur. lore fAsvs, Ch fed Gage D. . $e Tak, te ' be 30 to C21 tit >. ratoud wt NTO Te) tbe) DATE: REF: . 1 .2 3 4 | 5 6 7 8 9 10 i et, , sarc AY.C) > 44 “a Oe Laer, 3 ih (a> Awe 4 : ge ra $ V6 lw 7 H 4O vo gal, ‘ ‘ eevee 7 — + Chi. (ast chat, (¢ Ady ayeo >) Spt lkitis, LC; Dhactic Latta ety rtin br Viintterte? Coleus Urea Arann ort f Oe? 0% O 0 arn Qo 20 Cb ul andl | I prego of’ | & if te e S Wer DATE: REF: 4 y SE x “Ss o S nai ot tis ee ee Fe SS age ss N y i d (—- gs 7 . Si. y eee Te | 3r2 lo > 3 Ipdige- a 2 3 yr 3 be . Tey Y Qs it | i lat | SF 9 f Z | sda c |y | & | / 687 7 5 | (oO | kyla. g o Ss / [by Soe lag 10, 6 6 / ., 63 48 “70 > Ty . 50, / hve: - PE) ond ge Meet mes Ag Vee peciorer- /O J2/6XR DATE: JAN 27 1955 ——x SW-967 . FEB 5 1955 trails to agar [rt7 ne 50B—x SH967 (fresh). 1230_,40, | . motile cells fairly numerous- Let form single clones; HG divided= a,b. 5. 2 iw N6: examine for chains. Isolate as many motile cells as possible, and transfer these directly to oth motility agab, as inte > as well as mass transfer from residual clone. mm A fs Isolate # of Fla+/- Trails from pltd. Growth, tradls from clone singlechains. 4= 10° ~ TT ° A-6 0/4 vw Oo | B-6 L/h BA 3: 4 oT C-6 0/.3+ Y Oo D-6 0/5. So a IN E6 0/4. Y ; ro, (I-41 ) . F~6 22/ oh v F1234E34 22 Og oy H~6 | 644 | np 76: not law riany 7 sasrah _ b 1/4 | 1s oO Hee ages A-5 1/4 y }t ny Be5 b/ oh vx oO St 43 oft oT C-5 b/ oh / A: My Wee D-5 13/.4 Sy pl23h 3 éh3y, wt i? pqs Sure E-5 NeB- be Sr / F-~5 0/.4 G5 0/.5 | (2) G6 7/ oh oe G34 73 laa. Ke T- if fa we why ys ef ind Le ehyymatacte bi pie its wlbo * = 3o/ > SP ny / FEB BT 1955 ‘326 Capp Sie 3 neki, 1-48 “mia +6 2 pole T 4317 Vo def, adrats ~ plot co suh6le ! t Gl GL To (cerarers 21) 6 _ © Svan &3 f O | haa 6 sca Ulaf . | Gna) AL 2 é SF. : HI © > | [oa ly °: ~ ll ye 3%" O he &- 2 YW oO Y | y 2 3. 4 0 | ) t * { 40 ia? iS sa 4 cy 5 poe BY (ee ; of 5 i fm | 54 ; uy a 2 (un or 4 . oo 3 > th 4 | . 3 —O 5 Hib) / a 5 2 b i 3 | mA Z a eS r 4 ° b > os rh G NS Tae. : # jz{ 0 Mice Lo Sercetag foe D sts A Le att plicyd dp a laff NED, CM ce eteseen eh, Caan 20% Cle (EE). eat itl WA per , Sw: Ass. bs. bab Cha I V8 B tt va . fm be 4 Lechatirn FEB t L195 pesrdsl unk ford bbe: ddeonton pe G Me sof AG 4& Oi. FoF +E ( )- 62 8 untula fe 5. yh ~ akg winks 3 Dat shop tnrye Mesppat OCG vob ab ind koe srannit Might Wig dew sp pa nih ei pean BRLE WES Ww 745 [VPC 1 = pres toed! 1002 =fedovam 5 shoo’ BOK DATE: REF: _ 1 2 3 4 5 6 7 8 3 10 web wein dk z BLP me Suspend 31 Owe ow jou List WO = B} men [ee d:lhe | pio [=< 3 mia, | oS spite soos 8 #1 | plate. Olean lade NSA dite luae = 200F )R. plte 30% Hh plete Oo. lew a NSA ‘ dilute. tw = 390%) JruwL : (2A 12/6 te glada Oodce| obs» NSA poles Ie = BOK [rR , plats SY Fd eb oer ate NSA dilute HHO |= 30 yf phe Izy HIS eft) 0.0% ee wits NSA puis rio | 2 se Ry pW O3yY we ph Olea nA, NSO Guha | rite | s SO fw, | ght O12F #7 plte 0.04 cc ote USA dilute | nto |= 6.3e [eR 5) pitrilooss [48 plite o. lee otk USA 20 ? 109 /,L oma fl ->| oa, FR & Jo7 ele 10> <— 10 10” -— 16 °S— jo <-] 108 / wi once Oo foo om H+ 3 een dcte {on;wm, ok azew i) vae_cor.Q Bens ot 30° CL oft 2d~ 2-4X10% La Hein { t na, pebianed yee ma Ld aohy coleictin Br Cody ) rapdiceks acto EMB lnc ow prepettion ot “co: . Te | {oun Wer %0 9 Le A ) 9, 29_, ] 1% % 4070 £ ( a, 2 4 To 5 og | i 49 6 | 28% — ee iadacimes g | a b 272, 25 Vo 2d 4 24 tr 1Y@e ta L- 50 36 27% eo 164 | pe 458 47 | 358 2U% B13 174 (242) \ ; /214 50B —-x SW-967 transfer motile initials to mot. agar reps 1985 0 nelle nee re ‘The primary purpose of this ‘experiment is to evaluate to ‘addition of extra SR-967 cells to the explants, and to estimate the fraction of trail-forming clones per initial. 50B—x SW-967 9AM-1130 AM. Coneentrate mixpures and trap. (This procedure works very (SW~546) well. Its main linitation is that 30-60 minutes are needed to entrap the motile xells.) Al-F3 were collected to about 12:30, deposited no later than 12:50. lunch, collect to about 2 PM, and deposit Fi -Hb 2/mpmk} square) by 2:10. AE hss" Bihs, earlier isolates were _matly 2-cell (2/ quere) by Ca. 3 PM, transfer isolates, at random, to motility agar, either alone or with sup- — plement of cells of SW-£66 or ‘SH-967. tegen Sor Bar Tt FEB 9 1955 | e - Sug —_ Oo | | swab) y - / POH — 10 { ~ 04 a eo - f SOghT , las 31/7 cop Lt 43 ee voy? be oe Tee ; - | ( f so Ohas my sw7b7 | 46 3 harautf to Ander 0 Litrmetl sumer S967 melt vasth aq oul ii eyplac. , Thea tied Lath ey bg cult am ILITA whit 10 sevgla eels: tracks j}221 , } — fae wee tees ene . . - ee dea tee tte wee UE goer ue San th yt ie Met Poe as! pe retool rat) (0 0 BBE! 086) O 2). asa (ec tla ZO coll wn mudd "tow E= Nye sg en /? = ‘Ye, gas lf fe We Sraeince sero oY) yas Ths pt ore gpey Penal : Lon) x Se 967. j27e FEB. 1 6 1955 - vee ptf t pat . Wile fps PN fed ety fey fps, of "shale J wsae 7” C Hes cost : lls 92E Aly — FEB 1 6 1955 pains seth aek he path, page 2 ite) A [=]> Metal Colic gan, (NER) Co x96 plead Gling ttes ae | mr be Podiegtte) Stays rears AT A: [ry@to sabe WL a TID bys re and 7 wa nike het Weh -_ wn Ta tooo yadhin : So a he fr FAY Theat sine MN - not — eet do, | Ciseartypot , Mes ume. Ebel) F Ja eb, 99, Sob bat ©14 3q Sb bb. bel Te 0 ote A Go tr Angle [aie nooo LT be. east pu 5 doy yt tlm thay Se wemumnerprareh | D. vie > A eeeapeica prarth heed Aorta nk aot Ame PLILZ. awit? and Sto BF are [ ae | © $e Ove & Vote Aeplord uo vert ad up: _. FEB AL? 1955 Bag ook m (Aol ) 8 HEA sn th wb (apd (Ol yl famgelsn) Clos Cotaly fk ay Vee Tak ? SL tatep} doth. potBrn diye oth ol fact ep ( fad fe tes canny A Men 0 1980 A] yoo y \, v2 4S) O67 (225° Hin ny tak gape ent - Tink. rte tal Ol k butte . “Bele bbs forbey soe (rr . 7 Se Pry Qoctike § lar = | SID (eek )\rocebey. Peareafr te Of wR frat « Cabana raf dat a 2800) 1) Gpist of spredion. ( Was lop >) oh “FEE 19 085 4 1b Laprp degen Af ada of —> If ches. (42 2/ mn bgt TaD, pa le ‘ >) ie 1 1gk 1955 b elnusthud spend of a tee er clot b so svell disp varus dnd me eburtn qr an baad ett il (ln ad sane deb kE KS 15: nace ae (Yih wma es Haya eal oot Th, Ad dyugle yar tied an yams | OE plate logs pape E1215, 3,),0,22 AO cols. 79D si ss “pot hd, < gut ad porpard. 3 , 0,2 03 0,2,3,1,2= yu») 16 ato. 2T. 0/3. eee | (3/5 Ct is a (Az bay : oh ta 3 lips li ih neu moe lahasthaet) Of nL a “ye x 2x Of uk Zx oo . "ioe 0 / (OB uk calinkes { ae 2 owed sam Pa ~ ~ * log / Daud of, 0005, mL = | r= 2S. Py, of - oy . " . ed . Of. by Nb Rep. . ery We haa Yd Jigga, /2 26 FEB 21 1955 | My atlsed Hots elleg oly soi a > 120 (afr over ZO ml hath fn ase ag oe | Apued Mem UT Kae GL). 2,01 Saaghen crp2n1ss |? 37 : 7 rea owe Wo Hh difint Aweye TIC | Jw ss Ss = 28 = 138 (2 /,) Ww 604 ¢ 276 ‘ > fan gross Tere 3 z 38 Taber fp an Any ores A Pner an MEA. 0. oft «then He 3 FS | ath bra. b Ha oe 32 & eyelid euch sofas — te 70 a TRAILS ANE V, WEST WE % dpurd simh ¢ ca (0° SUG67, 4, IT feaks ; yawns he fy we hat, wth: $e Wot 1. Spit Mone) fot 23 1955 a | ce 1955 Pekan HL HP pa eee? Wyeth wut a a ae | ‘ ah ay jndindusko, T- U0 sngkeo G hath Felapalaynd i AN anusodet onke enabstf ull | ramets glligg ¥ murtren rele for mtn x Wiad yhcy > FEB 23 1955 7 ute ut Ma bel tl aundly. [ale oN oudeel? fore Tem jolces. tafe J ieneg ot fru asia af &1 Tene rt Aruplee Alp be week) 7 pL png OA hac / >) Ma igh gpl J comedcse | Spud ~ seetiches mn plume, Te wee Tog the tw pals SwIblx— etreer 31227 | B24-A 4 FFB | [heen pos. of - Fie tuo fromel be felt rome re lyn fot pod. 7 ,AaAyo® wpa taal. joel, —> A fotllad 2701 hbo 3 Ary 2D way Doce ik death leach. ~ 7 i 26 Shr [econ pray tla Mabe Ber A tabeo akg Y Messen (2 (out ) popes . Hyer tolls (etm. itd) SoCo aod, mere py tes (m4) bey Gl S finer te (1 ») oad oe th bf | 20) dns ee Veapio “/L27 FEB 23 1955 J -S° Py us 7(0tb) Suh t Seal FAS? = fof vn 22204 be. (exes) om cast hu “hon EN tk AR . — Ke Lapnodiag. Bq 0h hip: fou © fact cps DED wd ponent Shale FAe = 11¢) CTs Nelle > MH dashes 2 ot fem ee! j22¥ ‘FEB 24 1955 ane enna fuk 34 tuber (emt) radi Tacll bff ns tlm PL4-frs. fue IA - | bore. en v) hope aby Mop J pony adup m 6, free nut. UY. t.. sige al doen 1g hep a - ne . LZ wrirdsaatof Foe te fe cant EES & 100 ner . Hite i, obey ® gry fe aed ty y ae ere adil oh convened. A ee Hay "Ae coy Ue acovcb) abe SO -/o2, (49, 12 Jo (24 98 87) | Male \, THU canntt-2> [43,, Betlates “1 I in oe fon ha O81 Or 4 tly, C2 X ) ) 2 diag? — Matas jBeloae PA: danse aulle—s wane &\*% A Oe Yan pe sie fh ST 9 cola Ayaan ~s hl wt ‘per cont ff oto : oO te Mh crits ph lgaphd Ua te ms bo rly tale efit Pa pO if abi | pen bt theaters iu (hice ofthe: a Anes i ll mfg eh + Seine 0 ofr tuba ECHO, ay VEE, 1229 FEB 25 1955 lacy Ene. BP P= 0 - © bay Ge <0 [Dertdigen AT fk malo | a Pe. he HE [oiespel 7.) ~~ oP Platrdt 5° - set unlfpese. \ We Hine | Srnsler dota 1-2? , we ye pe - u “RMA € je phar crs 6M ang. 1G ee 23 Samp ~ 2 yor AL , pf cab] | , oll | ~ ae 1955 - nae BI. }uar poATL ny (yo - | " ' {| &. L tackle / } j A f } nage, Hols Lids. S| 1's T/C = 24 2 a pe TIO 12. >'s ( .. i ; a re “) ReurSe 10158 a4 TAR ILS 198 Wl 7 tins Spe 40 Hanae bbe ep eloverene nly te D-5BP2, : FEB 27 1955 WE . 5 lt? lc pag Peed -& [ oho s. pemucsel rumirtsde yg debgel, vulva 124 FEB 26 195 ons - ~ 50967 MTA 03 cole me sew ne, SPAS. “4 1 (nel | 1 YR ety. Wo MT, tetreaae aed 5. . A 6: 3) elms be ha facto at Wang A3- 4 siti ia ( ) vy, shed all b Lol 's 22,73 . pyr atere — yf ‘ 6 " fit a od tl pli nsf on — wt e ANT Yai ted O | . oe WPA fess Fak ina] fl WW tacts “oe FEBBo®? 1955 poet Ef ete ete Duh (sot | hesiudatef t tancinnef lp We 27 A Titled epee wie OMB 1-10, 17,18. 1 bw a Nya mo MAR G 1058 , Webd, pil Aaah @ ie Hatin 7 Sul 767 DATE: MAR 3 1955 Pa gee pacts tons SW 967 /230 10 3/1 50 , fag ty : yypnsr pul, | PLDT 07 oh DATE: MAR 3 1955 /22/ 2 3 ae Ny 27 Se, Ame 0 (24 . 6 l PSS ji32) Lh. ~~ —x Sukh be = RST, 1232 —» ($40 © MAR 5 = 1955 A. THime Swf fb. —lo/¥0 [peal C. FA D7—«swote, | Biman. Hanh ne lx. (e CR 2e%4,) bette @ ap -325) Pisbe G inl fererany kg Yor brah Re 7° ec, eR Eee, | Pile ne a Ey oe oe nth, : a “to A saves rte Tana $ a Sor/OT: | s ) 8B a Kore , . C ; (Splat " Gl GET en ellie he amend. panne A r/o: fh. ly wine fed wo Upon ns 57 Tart wiclste alent Me atin (wc | Sr 2, / omni om ‘ Nd. 24 | t 65 ' Co aly: bo ten 2 =\7", oh A # i; 8 in) ees, 2322 pate. MARS 1955 REF: Pied vidivnadof) oes all oj Sorell tO) [b pla maple 5 2S & chorus | Corint (>'s =2) J. Mhaifles dbrewk srl hudic bo Lb feadA, |litinel| Pamdy? 25 BO fer gropol 2's 73° if hurl d's 15". (smb hae bean dif foggy 2) e Ft faip (ner) 2¥'s J3 aj. | log | 6 1 facf ( non Mer) (3), eurolf Vs... Wf dnd pwipsuiviny) 14 hae md J's 6 ortanapef d's. ja | ys MAR 1¥ 1955 Ent CHS we ROO” 1E, v- FADD ( b . ~ 5¥ Bx ~— prretfren Fey yp Cry Leaf , eh fer a jd . old —a¢ SOE gat turk. /233 DATE: MAR @ 1955 REF: 1 2 3 3 4 ~ 5 6 .. 7 /| 8 o ~l, 10 prastccntioe ip) Banas tte (enmsayyl eatin gesth un aubidiase anep., of YE.) | & 40D. T lal pa. 3PM. bd.. tyae yg Wee | A e link empl nth teh 7 f 2 Sra pleas tanrliud & ea [2P/y, hac 32° GV (later d- a SPVy. & MB, ACH, ware j955 : ! boo (of CrinK ee \ be elattin 6.7 cui — el arid tif Adpare of 22 2) LP huetn Yoel A AK i Uwe 1 <7 A, i | . " Nhe Ueln 4-5 role. poe 5 . $3 b 17: om 7. ue dhire» 17° J. 9(, As, | C4) 1G | an 335 7 ofp 7] 1? Wo _—— — 7) (68 frorf erlried Qn 188 ; f, fe lee 1 to Fipits loo yor , t/a y "0% 484 10%) 10 CS 23 v MAR 8 1955 DATE: 3 4 5 6 7 a 9 10 - j an ay KT i 22? Yd mw LF : A ND BG eam Pot fo [8%4) Platt sO? - 61° dP ‘ore 7 I . , . Ly eI MAR 9 \ lone sees ¥. J I 20 ab: | : : ha ga fore lad ig ae & dhate OER & 4 oe v h OB + Re 4 Banle £ D OP 2 ms sh Ma | YG MAR 10 1955 » 4 )2 3948 EM x $35 $ . oe = Sm My ~ PGR SO y Fr N ~ i ape” _ “ow ag YS pon (iat ? Cnarat dt potties corbuges ? 4 hhh. et ay hn ha it 8A tah? Meaghan pel yp fete Cen O ff gt ae phtivng a Chee she id pack . abate abe twine arb be ge Sarenbeas pris | Vols eatin word oO ey swell [ 2) —KLG60 1236 cn pe ss eee he Sikeg Ree fm r./I 7 s . thoy GocLp HM Ge cath, 6 srgnsft alls 37 he | j ont pont ie Bate he po eg at by ote Ht I gave uv, tantly foLr Maer 4/v22), . 3 Llacen peal on nese musmtsveen wee " Theee ume J237 ee MAR 11 1955 | “8h . . — [wen . | Lb tah — : . : | a sd OT ¢(2,3..) 1, —, # shod 20 2 F238 BT | : 3 *F o (2°3' oy 2] 20 Bs 7 yr os 2 sf $49 Pid? —w Sh Moz. as forston8 Na 40 855 elf / L32 big cl pbo (dat fk < Phe i nat) A. Fesckepoit=. ddlele 6232] $2 by Wan, BD) Dawe fab) (bln frat of HL. b bale pooD te rte. 7a Auk Lule (= 0,%,10 cll, on Hoh fs fan Se 1 OSop4. — HP lament tx f° tad Le one Sass, y Be elle - 2B he oar. » eibrard: So fi '$ ce ie he . ° i MAR 11 1955 . ‘e be 7G eee /. | A, Conteh Mod ue Ba, ot Boa ff hed seman id howe? js. » haste 5 main eon len, ng il 548 ahr most may 7 he [2 T: dp oe TRAILS i, BI, | ( at * Basdhitids , at /7 Beg | OF — offo Leatuwet (03%/ ) fs - ait Sern ¢ atesdinaf) FI Nove AN Sb alse “dae sonuve 5 ules fe 20% mine / [orekt Mn voeehod Bor catinl e cope dur | J. o | [Oo 10 7 \ Tt we “ / we f - MAR 1 2 1955 i (bb pipe? Wo troLe . MAR 1 2.1955 Miiabed © d Sd+4 RK t6- Cre Usa ef ; rites Covet i 2] +6: fe “ate e antonel , 2 ZS 3b Y- a | a OFLVST- . | | | | 2% ye TH. . (27- : ae usb br flaw - ff fn mebatit peecleg - Pexbicibe ap pts ~ te se Sofeke Sy. de, aabtionr (oesgpinel MAR 13 = a - pipatie LEG cay stepuntd uly? thes. Co . ye Th 0a tf wmted. MAR 12 1955 Bearings: — Since Jan 1, I have been primarily occupied with Salmonella trails. An important ques-— tion-has been the uniqueness of the"Z particle". This would be hard to establish by quantitative data on the clones directly, and I have been principally concerned with looking at platings of small clones in motility ggar. The xwmekexkk results with SW-967 are not fully reliable owing to spontaneoys “minor trails". This work has been done onay since Febr. 23. Before that, from Jan. 11,I was mostly fiddling around. I mist have been preoccupied with otherkinds of experiments __t00, or weiting or what not, since rehatively few experiments are recorded. There are also some experiments on crosses of heterogenotes, but DCG did most of the routine on these. From Jan-Febr., there were a number of misc, experiments on conditions of : . plating ete., which amounted to very little. There were some indications of major and minor trails. Also developed technique of trapping from conc. cell susp. _ Summary of clone platings. . (trails per clones}/dfd per platings) and comments 1227: 1/20/25 T: uniqe ° —x 967 12/60/74 All major trails unkime. some djst. mass pla. 1228 3/9/10 : majors unkdwa, noticed addl minors 1%/86/ 100? 1229 8/31/39 All T unique 24,/198/200 OO . 12290: apont minors . , : _ Non-linear however. Confusion with | spontaneous minors. ee ©. 0 at ERIE caer ree. —x S666 “Almost as prolific source of mtiles ~Izgz «4 /18/25° Single mors, pot otter clusters. 2159/22 * Few, 5$w/13/79/100--- Clone size c. owe a wn 1233 1/34/47 Single small trails; a few other ——__——--— - -glusters. c. 26 Lye a ene nena 1234. 6/25/48 Definite concurrence of smaller trails or larger 30/149/200 cluste 33, hayd to define. Av.CI.S. 6. 23-4. 3 sw. 1236 57 clones, follow micrgose. plate only 3 NVG. . 18 el., 7 aw. 1237 (eff. Tween) This prepn. seepsng. 1/ 173 fies | wo ee a although very fresh. . 15 sw. — Should compare directly with 1234 prepn. __._48 clones. followed micros. 4 gave. sw (segr. non mt.) ee 10 quédéscent clones gave only singles; 6 with fairly numerous motiles gave 2 T's, + 1 with clusters. MAR 12 1955 The principal point is perhaps best met by experiments like 1234, plating fairly early. A corralation of trails with pluricatenates like 1237 might be worthwhile, but more laboraious. In view of sluggish motility of early log phase cells, this should perkkps be done with earlier clomes in aged medium. In some prelim, expts. yesterday, I noticed that TM2 transferred to aged medium supernate was more actively motile, particularly showing a more jerky motion with shorter fgee “path. Examination of TH2 in motility agar suggests that many cells are directly immob- dlized, others move in apparent interstices, but still more slowly than nobmal. There must be a considerable accidental factor, and cell with numerous mobile progény naturally has best chance to propagate a trail. Since genetically competent TM2 are immobilized, there can hardly be immediate correlation of genotype amt (or plurieatentation) and ability to move. Should watch traiza in situ if possible. Why not? Plans: continue with experiments like 1234. Compare this prepn. with 1237 in yield of trails. Continue with medium effects. Set up trails in situ, selectédg cells which remiin motile in gel. to see if these form the most chains. Do not forget many other carryovers? ~ 7 G&C - Gal, Nelson | . hetehogenote cro p) GAN Sabot, e ogenote ¢ eases | AL | J Hfr x F- DATE: vate. Whe I ( Wed )_ ner. 4 | 5 6 7 8 9 10 War Hepa: (an —H/ ea disyar Jd >= ee cll. (520) oi SHR PURE > Sal Sirigo t. 2S nf). Fgpucds rl tte Bok Zz |e 2 S323 2(¥,.) S | To 2 = Agrer>), Pe scp (¢, Om: |S HPA ) ted bef NOD py he Le grleameb Lliccharr ( lh >) gh Feodeaon MW 4) he i fb clnunealf, é es .2O YA?22/ 557 I2394: (03 G/ /Y0 144 67 117 4/ tT /§/ 37 Ns S uf 92 19¢ ST 1Z2A3BEODO. 22 3¢ 4 Oo /s S 2 13 44 12398: 97 ——— 225 134 leo lor Quite tuaks “C’ 3/ DATE: Mase 1G! Eat 7 Laet cor apt A Tod Ayal. Aaiglea. Gre et, D- NA fer) | 4. 7 dn tly, op J. ueA (jor). f om 56> ’ one i i tilayr Mc ap lel —| 1) SiHyptttats ¢ b hbtin . Z L . 2 Y , Ay TRE | — f Wnstt®D 230 Z 7 L iMCA (pes) (9 Mak rae h ewes) Z E ge . M . , , Fe, as p 42) ie for ) ¢ canagtenY etre Let ® tists tc eet po ff pe fe | et a a A (Cf w), | si ' pL, ed hee Gres ; Sava IK? Avtef soos ? SC , ci? > AA f 45%) Bsers (FT, [Center Ss Bs fib pew ‘eget (a) 7 | mons (Lag an tae ank in lin i 5) Aly 4 a hn rune, 3 oe hyve h auth lor ca. \F LL cle AM nl yon Aayal baal Moxy be Anel te pidigey ie ‘re ul retest , ASO r iy hot tl GMT chet 3LHT*) + tT che ele TE ce? S| (1 C ; rr 10), qrac}| & 6 ( (63, 74, Shae , Jee, D.. | 24 “fp 4) “ “yo. yA 2B TICE ATP | °P 12 ( wif 2) bo. [t ’ i | wn mg ops —~S* t) AN lv, ager eq. lo sage Ges bee rv / nee / )* ; Q thenle /O haw j, 5 slat hackle ~~ Ick; af Cal t- ie pe bs" pe oe 5D ener 23 wl 5 » 14 tachs feels. 8p te. Wick ? I Liman O 1) corks @ Rr, t figchy . OQ teks ] Conclusions: 193840. MAR 21 4955 among motile initials plated 1. Incidence of trails/varies with the fluidity of agar. Addition of 40% diluent gives very high incidence. In any event, agar that is hardening tends to be quite inhomogeneous, if maintained at critical tempseature. 2. Single clones can give at least one trail + large clusters in harder agar, and numerous trails in sofeér. This is clairly an unreliable criterion for singularity of catenation of higher mat order. Further plans: 1. A few more tests of fluadity and related variables for photographic documentation. 2. Shift studies to direct pedigrees; need some further data on irritants; inh. of - cell size growth and chamotactics. . 3. EM transfere. ales Tal douoll mohsne. / Bag ‘.. ve) hal eprint sdiglar- heater DS - Seams vi oad tal Mi prraming yn” fafa Ln nok flow wd, prdiyerd sro Han 3- fof rg ble ab allan, sate “belay ed Tg athens O Siam ofict ( Sra Geren milf my | fy . bg wore) Of dat: sl, < atifbesnth be, te bie. (omithn ot) APR 1 1955 Vharede Arann. [a Hf ce LMA (E a?) ~Xb66 wad : (e) Vor prgn. Vo fofeg =} fo — Jy!® fps Ee F (hlitan bh beth» Dio) / cg [LOl nh Ob Lp , x , lt 6 Vb - J2 Y/ hf. ty 7 ! Jost dy aged |-Y. | B ‘ . inary en 7 Jo 0! da eon (i (b a Cie) bah 3 furan mea! A<& @ 7? auth forbs b b 1 | ferrencouy ax SH 666 1242 30 | Penassay gives at least one not re - 0%" broth for plating clones. @14 had without further C5 had (Ine. 2 about 29+/1000 C13 growth. 10+/-00. [Plant ind 12:30 — 3300 Pa, Pl - 8) 100 Fla~, (42 3 |had 2+ 4 66 12/. ividually, pick td ate in MGA-40). (Spent broth = overnigh refr. for aml yes. (dil. MGA very soupy! | and decant. Numerous tine i still Note as many. 23/1000 t clones ) are limited to 18 clones (in =e broth czaniaeds 5 wd 1+/c. uh, of which 4 isolated to él, 2,3 9 each teol. . (Tange wicks le + broth for clones, (es- 1-4 vtag Fi sent). > While | and plated be 2/0). pate: APR 5 1999 REF: | 1 2 3 4 5 6 7 8 9 10 New . a): from aerated § » b) from unaerated culture. Both #1: 1 aiial ps2 11235-12210 Refr. | (Incub. in rot.) Note: Rotator fo ! now standard for -asration. | A): Prepn. a). Fuse traps 12:30, collect ca. 100 motilas, but use to spat (2) . 2~2215 ‘ 10) = BCG! pick to i Incub. in .5/ml Ponaspay to c. 6:15 PM, Plate in MGA # 40% KSB as diluent. B): Same collection, plant in spent broth (SW666 Aer.) to of. total clon size. St. ET. ch: Prepn. b) Fuse 4 PM. Collect 5230-6100 (some needle tr.) This prepn| as least as | good as a, probably better. 1000 (+) / 1 ml. Plate 0.1 ml samples “ire Plates; 205 in small. Compare 20 a MoA, +4 40% HSbdil., + 60% NSA dil. . 37 9 7 n RT to ey the sediment APR 6 1955 (1,7, 13, 14, 2% B I 7 7, 43 ) 3,54 >7)23,3 4,97) 307 6 e's 1S -¢. bo 1's | | AS 7 . i's 8 t!s (3-6) iT p7? (| Y oe 27 Image fed 4 qe's (a7) + 6 /-2's. poh Y, | 20-39 2 ae fucks Yaad 5. | smuhel ketal z | 4 . MAN jo 3/50 Ad's art ame 14,24, 30,14) 3, I), 192, CIV) Sure bv hr , (ai® M4 116 Aud lice | we 50 m j « pouhei). ) 37-1 5 S ¢|(<1a) 2) T's. 16 al BE o's : 18 aamesomerh Mallen, \2T 5 luda (3-1); 281 /2Y2LA DATE: REF: 10 ~X 666 )7¥2 37 7 _ APR 5 1955 () Bor pp A fem acalef iid) iy ne nauld « Aa (Sime) we Pe lhe yn of a a than Pll) — O. Fase dimge & * 7 APR? = GAM ‘Seme gous Cc, ¢/ 2 A Jo I's ; po chia, 7. | fey ole Boe fe ?, &, GoOI'S — , o34 dic HUOvs. I. ¢ fy Mir Sane | . C 3 a " ~~ dD FY CR ¢.30 15 le C - O } C- 2. Yols we T. Nehuwr 0K. : 2 DATE: APR v 1955 , REF: 1 2 3 4 5 6 7 8 9 = a x. G) +. t eS o\ eS ac Q ee Glanaan cleertic y #), an Ja, Oa 00. | gp ob DD | | 50) oD paTe: APR 6 = 1955 REF: (ap RD Tal, Mar, ok Sane par m , Cc. is (Sud S Bg |G TG SE Sy 9TH, F ee 129 O° fF yz 199 40 50 H - c+]{ D> wars rutetated . LY/YA 1955 APR 8 REF: DATE: 10 Tus Add ef roan he tY/ . _ 1, om, ! rm 5 > > ‘ ! » \ : 7 Pr ay S ; \§ a) og Qh om] © pore trredce robo USSR QRS IFO R20 Soe % ~ ian Shy 23 | Sts | | ff Tze yd — | 4 a r~< 1. 3 Ths 5 “sa . afl See A-09 1 nS: | 3 Fo Ya=-4) QO YoOSe wood Ff jo 00Q90~ > ss | IN 4 mm) : | ; ° ap& | & a PN g ® 1 > | Y Ss L | x ~ v | | | < < an | 3 3 » | : Q 3 > ~ a PrwWswr Seo tach& > x n 3 i rr APR 8 wore a S) W / 1 2 3 4 5 nod Ye = 1 3 10 VOWS 19 athe 4x phn fred fect [0+ indecle be iy opt ni) ee ape eli Bt be, bean ot 50 MO re m feb. | = 3 - é EN4/Dle ¢ ote Xe, 20 f pyre -© ~ 70 Lhe. : 1955 REF: Ay, a ft sn Je 5 7 8 9 10 We q rn * 2. = ] hae tw ATG Alyce an foerdes ‘g iy tT K ? x ba farf boc nthe ye 2 ee ly Ne tre 273 S, 4B OLR fe Iho cee oa \bAG NS | ft E17 -& UF Cau TN Ahive Ae 4 20|| ™ 4,4 ctu! By punto | We, THY bog || OM Ao” eth + 6. rr #3 é ara 30 /v 3h 2, / 4 | yr¥2C - Oo - oO, 2s _ be J) i Ya r a fun 7 Ne 50 ; . aT a8! sa! ¥Adleyup hd . - ¥ OT te 38t a oe 15, 7, iT fo! aT . * BT sc so0's BOD 7T 10 3BN's ee BE IEE MT 334. att AIT $C 371% ( Cow X tanger z) pl gels, Bot F1* 327 and ee ee ee AAS? A ¢ YY 31 2. a Sh . APR 9 1955 pyre oT ne i, (ele an | Moet Moa -2 o kelp wT. Yo,be ieee TS, meaery yom ae ; ne [vf>. bapa pg, pets nuded» Stace fterron, toy Lbs: (/- 6) jul (ebb be | foncsee? prettier. ith tile ptt ae MEA fo- So 7 We Ste clones OTS ye ry, “pe or / ee Jaw - oR c y/h0 S)>7 (ee Wan, elle, “tg ag ae glen e woke Poo ases Fact pet Av ees E ptrse Itawt eon LLY b Hanns mt tant of Paty )? | 7 aa / / a | Wh vig avr bea a \ ef k+s] uy | elana ‘APR 12 1955 a DATE: lise pe, dip? Joa, Wal, whudlin| top, Mia a> dette) we Crhepezame arectthe hor fo rtdal ire bo Austclhe Dut (tea "spot Loe H PA ol fallin wv DATE: APR 429 195 2 10 40 J wh . 1 deta We T, APR ]3 1955 ' 2 2/7X 666 5 “s . | 5b? lla NEA eh 60 $e juiver pale — KPA. dhe «conchae . Bucy Go ' ; tee A thir h sw . , r r y coitef swh73 3 Sy v = Sy > > 4 30 aye L , an . Of 4 . APR 14 1855 My Ye SO whase eo an MEA bO | (Ot Weems i | . + —_ + tT tT a 40 pd D ek | heaufte To he Fe APR i 1955 . j3 | 1 ae ‘ * ro Seuum intelli, 0 -* l2VP pare. APR 14 1955 oer. 1 APR15 1965 5 4 5 7 8 9 10 1Y: rn t/s) Facee deme , FATéa C$. mesina a) —» Swhee WAY FAB? —w 7”. , ° are wt bhp bt eettawbict fat a havo nth b fake, pact ah aff Mele , bankht all poits § ayn fy hee E60 urellRo 20 18/60 farce ae 43s —ssenemes Flat: ,° (gt) ) pedege 3 | TR gare usile om pamfin, froin nuied +[—. . | oo (eR H,? +) My tall » Wiy alf - N2% Leeks, Ulrumael doll % oln hy: 7 y Jhourh leben G "3, Ns 8 :) dscbo bith stl, etoustl © WL) HW5, ng Atl . Dis. )~3 oy ¢3)} —> ar. asl, Hee talinale) (f32— bee: tf let tt,* ry) ' Lp by oma mxed Fle th,* Sexe t hy. _ 2) Reells > ny 4o = . | | (nu bay 2 bare) 122) ) | hain Mpg Sattee MENG hg <3 Wd charino (ro > 4, — ng —He Sn, ) N33) ) Samu t pne (Wd mntle, Wy ~> 4, Dot bli y Mo PA, Nn, > Ay 3. (2 meters) ee ) Non, Pha,. $< ro+- mo - mi frebs. t - pe) nim ts S — (3 sali ns Of) 4, > ny. |i — ny fou ST Hola AT ng —— Nyt chan (Fy) FPream. El svame, bo, Marden, F , 05) WP“ o — ms > 33. Fjrotelc : vy ng ne we we ? rol es me | 16, 5O whtrnreel . S . Nine, M6 : % mony ne? ust cnrtel. La le _ ne 4s ? N20 _ | GES a > My Brat. Auvth(D ® katho— jel + 13 Me ny, a yo p23 py my te Poa bi'm. f\, & a ey 7\) N., jor t 1. J 5a, Gt chan rhambgihe Ad 44, AP 2a 727 A 23 17 2744730 Key 32 WY 3! %4e 22 23 24 22 € WI 22. E 2 7 22 B22 32 19 par ay 19-724 I 33 > 38 CU, jb. 22 EDR 19 tz 4b, 27 3Lr 724 c23 (nl Lew Fi C3346 5 2 ae >/3 3 cw 13 Aeey 3] 2 Dw S iets 1> A>%. 72» Are 313 3. > 18 > 10 623 j3, 337 cS wey 3. > }3 P22 13 2s” 533 /» (2,8 FS / | 3 < 6. en Hy —* APR 20 1955 DATE: {25% 1 2 Faqs (s fA? |. {3 x ine Ai 37x | APR 2° 1955 Bl! oo 2 td FY0 ay, —— Bi 2 SC/pl aw foxy~ f pny oF ond ne ee w [BY paki mk > Peep faite? ohana bf{o B b cP C ke oO a have bul Bar puuthogee ere 04 . a le nab bon pam f A et) 6/4. ACG ; ™ ; Mn? Nowe metab, 37° wpe wok ry mS tt 4731 Med) . , | : . b: poe ls c one ) cali pe pote [rhe y2>7/ ABBR 2001 19955 REF: ME 2 | aad 38 © rea me lf baal cu 6452764 A112 i } WW < Ox a Uhrabe for roll,’ © Severson poral D Ctra) etows't pk sary tir [inet > (teak pO Ome oe HOt rf brug ° 10 27. Vy REF: DATE: (C38 _ \ 3° Sy OS Q 8 - t > a — ‘ i £ 3 + a 2 ® = x $a Ss a. 4 7 - a x 5 Strath Nom Pat ocbe 9 ee POLO STM TES ys TRITHO BS Soh booa Seater: - | : e12 . * 38 Bl garo=z 0 8 oF BoNSo J a & 4 : ; 4a - 7 aa Os — He oct 37 . 5 ne Ge SN ox foot “6 = OF xe Oa OO S (S t Set SP elee OO =~Ca ite 7 1: Of . a ep Yoote === +E a= “Or ©) cHO OS sayy ~SN S GS ANG BS a e ( Jmf) (dt) GN23: tals co A b-xb ++ ma | Ls sat be reef anderen Fla, 18e “fpszar cat loro t os foy- tyke es S % 1253 Tetrazolium- formazan chains preliminary APR 3 4 teae 005% P22. Prepare stained cultures. Add 1 ml overnight culture to 7 al broth + TZ .O9f$% 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. we toerprud, Zo atl drat 2 Fate bukf ae rag wd ae Te a5 bok Meto 6 wind oe chil 9 a edt , oo —>» eco <> @ oe —x SW 666; divided clones, TZ 1254 DATE: APR 25 1955 REF: 1 2 3 4 6 7 38 9 10 5 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 4 /\ aotetlnr-2 : | fark Kanon rgpeta line . sy Sh A, DATE: 10 © | | o . 4 $ : © > Woo < —~ , ABC ost ‘ : a Xo . SS a Qt 4 | > * ° We | 3 we y . oy . YS <0 § a ~ > ~~ Ss | . +r DR <09 \e No ae Soy a Yt < o , -s 3 —» \ ON xy S&S ® ~ ec _— Ns s a. fs 3 | x - 0 SN Ny me fo “ @ ye yo 7 > 7 LoD 4 2 ; j BQ sO 9200%0~8 99 sao ogo oo ¥ 3 & on Q =e mae RS yS> oa nm 4 : 4 ~ A N- + —_ : ~ a Yee en mfr NY mbt Qron oy ~\y N 3/ f ™ .s rin bya OO TITS > ¢ + 3 3 AS y fe te ul s . “ie QO g| fo FPAS~ O__ gv gd THK ~~ {9} \ ey, ) rh eg SARIN SESE O RU REfy URCUBU : = ee ahi ! D S fo 9 3 ak Rae y ak _ « y yh ze 3 re 22TH OOF th aint FoR ERO SRT OWE 3. NY o Py ~ 4 oN > ‘ a @)N ' x3 t So ea OY hoe gk i Of . a c \P A f- aA {s S * KP . t ry 3 ‘J , 3 ° 3. aS oS 3 AY 7 is ~ nrg jones pees pant a an — 20 30 B), ballots, G>—K Ka LOE Jed tod afr Abandoned, Apr 29. anpmnalies, Cone. Mo 40 50 | Need: ervatio: should also | 2 P¥ stained W-1177 (c. ‘Plate out in mthocel broth on c.g. t cells stain unipolar. Rat terminal granule, 8:20 ~ 104K Owe. calls lyse ns at first diviati j / spot a fair number » over of re (5% bi in rando | Some exceptions with interstilial [antes pn of su 1. Also fsolate a polar, subpolar). position. ! lar and of unipolar cpntrols. >’ o Al ¢ i i 2 bra... mixture overnight + TZ bro tn), | | | | | bipolar cells, 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: nity pO I es 12% Hatbcef Pas. Add ellos oq Bad ‘aud th LA Mg Te Hehe Most chains chow f2s>_ 2 Ch ° WY Ls is ye ° Nad . xe ae “| oN\n 3. Ff ca co | * | ) 4 BS: x a SS ~ a — 7 a x | < oy ox " ‘als = S eo 4 op \ a Tr ‘\ i. ‘ < =) 3 a ‘ x of p 7 , ester tleaygre dale cbr dy Corea IE y aoe EO AGO yla cq EQ H%Q KH DY OIFN YO. — Fiat H,> Pe A%—| FAG3 h* PAR qx HQ ke Suan. Gx=b\ Hi pel : (- Wt tt Gurtpee- i ~ Aulonste . z ? Novevain, 3 “ " . SON > 30 | : | | | . 257 WHIP? DBehumur }257 DATE: AO 5 St Davies allo orp: “obs x ty Hy bate us HOB Ybacpe | A Ma Mie adam [Hele Yarep> gin ren the sox ps 1, Currently enmeshed in bhe fate of £ granules. Can these really givr any importany information? By following a granule during the growth of a single cell, one might get a clue as to whether groth is interstitial or bipodar (jn a few cases). To distinguih, one might have to show increasing separation between two granules, before figgios in a single cell and thismay be difficult. It is already clear that 1) terminal granules usually remain terminal, and that this is the most common type, already suggesting a polerity in the cell. Occasionally, bi-antipolar cells are seen (more commonly that bi-synpolar), suggesting that the two poles share something distinct from the fissile center. However, the basic interest in the Z granule for the current problem is the possible correlation with E, and this, if anything is what should be pursued for now, Later it may be conveneint to try to bepeat experiments with a polar-flagellated organism. Another sideline is to use the shaihs in stiff medium to study other problems, chiefly lethalktym both spontaneous and UV. Also look for data en growth of branched cells. (Twort) 2. More pertinent: 1),look for divided E further. 2) diagnose E,S sells by viscous media, 3) transfer intermediate chains for electron microscopy 4) clean up serotypes of co-segregants-- collect more? 5) For 4 and others need to complete review of data and write up. Lab plags: what to doS? Things are a mess, 3. TODAY: Clean up what is accumulated to look at and photograph. Start new preps. of 93-——x w/wo TZ, Use for divided clones and for Z correlation. Motility selection: initials in methocel 1258D (Sat 5/7/55- Sun 5/8/55-——-) Use TZ staimed prepn. 5/6. 12n7 Chexked first with 1237A1+ for swarm motility. In this series, used 2% methocel 400, diluted c. 1/10 with penassay. a) use methocel for trap; b) isolate initials in broth trap, then trANSFER to mel. The latter was found ineffective (probably still too stiff); By 4 PM, had isolated 13 cells still sluggishly motile in mcl trap, and 7 addl. which were at a distance from reservoir but not now motile. swarm cells were sluggishly potile in this metho- cel conc., about 56-70% were directly inhibited. This oln. probably wets glass more effectively, at any rate it tends to spread, and a few of the mbbiles below may be contaminants from 1237Al+/ The motile residuals above were geet in individual drops of broth for class. as Sw. or E cells. wes found, in first group: second 6 swarms, 3 E, ang, 2 Ff. 4E lng 2% 4% which demonstrates strong selection against ~ cells Total. 68S TE 3ng Desailed counts: growth motiles 1. At 9 (104) 2 3% oS swarm, 50%? - 3 At 2 4 ong The occasion was also used to 5 At 12 (from Z cell, but Z nf) plant about 25 single motiles 6 nn $ 0,1 (removed before test below—per- zBp 7 mot cell haps should have been left in it) 3 20 as 16, sev. shakes, prob. aw for opngcoddance on immediate 9 aw ay and later motility of dividing Jo sw uw chain cell. About 12 usable cases 11 like & no discrepancies, some to one or » 12 500 eS two later divisions. As none gave 13 200 aw pw two mobiles, pres, none of these werexE , Of remminddr, most gave - 21 it h two app. nm at this division-- 22 ng it may be possible to reexamine © 23 he 18 these drops tomorrow. What is 2 it 2, significance of this crisis 25 ia 2 in termination? Is is growth in 26 da 15 fresh medium? (May still need 27 It 16 a good exhausted medium to keep cell size amall.) PS These were then used in tests for residual motility in mel. Unf., 1,5 were wasged in 54% mcl 15 (calc. vise 200) which proved also to inh. swarms. Further tests were then made with mcl 400, 1.3% and 1% (1:1 penassay), the latter being adopted as it permits almost full motility of motile swarms(from above). (This may be too. fluid for accurate discrimination against ¥, as will be saen). From E: 12,23,24 26, 27, adsegether cells reisolated which remained motyiere planted for further classification-> pr ute Ewes. Sven pro tere. a pubin pales ! DATE: —x SW 666 initials in methocel and Z chains ba serun. 9, 1955 REF: 1259 1 2 3 4 5 6 7 8 9 10 D: 20 40 50: Pres Note A. isol,. B. isol. tr in 6% d peepn. (alec anetior preph. unstz (1:1 & 38 isolations, 3 ng; E-clones were r able or riginally The jclone wa 7 hb out used. (st 2 9:45- ned = A. ye out of ) ined, plant out erate Z-stained initials. to c. single drops on isolation eg . Ditto for A-— p T. are inhibited and may ones. ed for sferred clones looked as if o initial cg. first, aps: pick ce. 4000 s did persist in DCG for results of platings(after but neither viable. had been separa swarms; onjy 4E, none interesting 233 43 5(sw). The therefore certainly not BW. celjs/ vered (owing to drying out LN; phage to 1:20, centr. thocel droplets. 3 | | droplets individually. : PM, some t out descendants initials | (somewha aorb i I i ve part once or twice at ntent. In B2, sib| to swarms motiles in E2 tes | | ce. 100+/10% but to veri and refr fresh isdl. C to 4:30 transfer latter as families bo t late now jor tes serum, though cells may co » planted out. 3 proved viablep swarms. picking to bro th) dn dimikashed therefore. 1% Methoce ‘Probably too thin.. norn Ae 22 motiles, | bed » all tse later proved , 1340) isolate initials ih 1% methocel 400 tsJ ntinue to owever, ot gave risé to to be ayrae fy originalY low assay. {2 Z, D PTT Ok / Otherwise ! | Fe 5/10 interm | but 2/4|\from B15 of se should DCG found Dl-3 all motile jbut with "all clusters"; & pure non-mébti numbers /. off inte. & serum: |at least) (28) from 5 clones were DO» However, two_ 8 were inhibited; spctfietiy Also saved 1259B1 (= b8). |Swarn- vast Det ediates were not sted swarm a by piating removed at ns = nonmot,a) | confusing clusters. Bl:. no le. Will have to | be recghecked on recorded, (not not cer definite eturn | record. A swarms H2b | E: 34 isolates planted w/o lineage afterward. 4B (9,11,15,16) | vF (1; 4y35151,657,35355y55251, 351545 --00) oe 7% 2 Dg. Only conclusion; medium not adequately selective. Try 14% methocel 400 (vies 1260) $f/2. Metd ow HbA eee O/$ O02 Tt ae BAAN | + laste, ‘Ds Bi, se Bab arabe © elnotine nbs, Cunt ! Chuskine © sevarine Saabs = D/ Ss! 2 pr GI , I D3 sf a” 4 6) 32 2] x B2b IC 17 ¥ s/t CU "anche “pg hd sVi3s¥ apeled mm MGA wy thi ‘gee had free 7" nln eh Mew aun) ame Utd agar | Qik ov gered Ieee, wd, 3/13 (OI, D2,/; D3,2) B/,2; B26;2.) Duele Shes ch 87% met Ba fn pt Arbt ethan f s/t. 5/16 ote COE Cost Senate , trlind anvanne, ¥ eh, EF VanlDhte* s € Oe = e4, deyhur ates » 7 D3 ~p) 2 oe OG Bi Lacks, chiolire, oppareh Borghi MO ALO Ae BRx fer nee Ag B26 Ge chuclne Qritmd JQ -: D(Y) 46 hule. DIQ) Chuckie, cette f merle D2 ual, tials | ant On gts D 30) Cuatro Anand tn D3) & % fm ans B11) f Claatie, tds o 1 catslidis | poritly anne oso | Bi Meo Audis. HBR unto ee po i P* pete WACDE by abe. abe. Zeoramn 5E ee Cpl ure Loadudl, sofas pathy | may 1 0 1955 No WA sane Heese Aura WA wr) Mh ome. [afaby heal wae t Dr Swen ‘Las te / | ml Jos, | Oot a . (e ° Gi] KI Yaya --x SW666 in metkocel 1260 Partitions . May 13. New prepn., unstained. (probably usual, about 90-120mins. ) Fuse drops 2:30 Collect to 3:30. Cf 1259D motile. No initial was nearly as active as 59D. Pick those hhat have mowed the furthest, not necessarily v. active now. Estimated yield, 10% of broth yield. Noye s to compensate for spreading of methocel solution, use cg. that has been greased (human), then flamed. This workesyd¢lwell, especially with labger drops, but smaller drops are too convex for best visualization. Intention was partly to look for early chains (E) in the methowel, but tie did not allow and most isolates were made to broth directly/(A, B resp.) Lineages were separated at No-3¢ As 1,2,3,6 ok, Partitions at nj: 14+ 31 635 ng snakes, Later transferred entifte clones to get fullest estimate of motiles. Al came out +(14):6 Sepn at n, = B1-14, 21-36, 4.ng. Mostly non E. Records show at first scanning: 23431 Uh 2:1 3 7320 0354;1 13338 3 2et 221 2:20 5 8w3 sw; sw3sw (1260B33 later DCG verified purity of each). 6:5 7:4 Underscores were rechecked (on ungreased slide!) and felowing definitobe Zalues for splits on theses 1:20 8:20 2:2 4:12 3:2 7: 26 Therefore no equal splits. General totals: E 5 ng 4 sw l E —_ Little if any selection for E in 14% metkocel.400. Need 33 2% which probably totally stops many motile cells. ee 21M sI\8 f x/24, (, 2 ; rly! J26] 3 4B Ligchatinf! ngtaler ia CoP hecunirelin oat ‘) HR paced pod sprilad shies ux (itt, 4 wn Bhr tte Ae en, TO lig oh Coe A | cee eee se bh , dec 30° | ) annals fo? 4. . 426,204, 4 2,50 3 20,10, 3,20,7,11, JUN 2 1955 © (2, 70, Seerdve | , CML withad ro Blas dybeor bcmrtn, OF | Wiiscore , Flat (12278)*) wer glee 6 bem (70% ?/) fod we vall, tif to toned “hare oe Nie — CMs part ened, bn An eotnuty furrle manly ptf ale nso tM Swan: maurkd ebonr f Lene on \mal, Ol wh 44 saouner AD fin Peed ‘el PP nb eos | (ove poke - phot hed prom hd ART orem ume 2'-heweo ? f > )26 2 EGO ey ie oe a ery pty. Yo ve fA lb afer oe Pe adaufr Ef, Hoag. Frere mky BE ehones, AZ B# a ow © ,| c Clow, \ ‘ Ae Sen AOS : | | 6 @ . ep lo ane Isa 3:34 awed 28:0 ( bere, By , 2,25 Bur aurum Aone C4 eluodapuat.. DCO ehuludl panty (Sint lege A 7 | & &® 5 Crt | eojuArel-S Whe =P nd Do Ly thule. poet Howe PEAS 2 anhid $( el ftp apni At By! Wukink 38 Of ) x swt bp manastod tad, oof |, ~> Yq. Wee ea EST ol Vd) BY 1/9 N41 Aa Vip. AS 3 ? Loy sg = 2 G G H] BZ). | w boob op. B33 O | » | 34 vp Al,A3 OF,01 ng. (ged mat) C2 4 | I 2 av. oie} X-32\ 2 4° "O fates ! 16 16 4 aye any 37> uke puns ron | | 4. Yo Yy nm ts (men) itel 4 C. | > uz >_ ‘t B Daal | a Ini Y [oA cama rsa. a0 OA wary DL : | 2 27 Ao be dukd og ofm'34 ! BY Y the lols tur Ss Ws 33 47. SB? ao LL 3 subset = prt A Mivd bunebe wi the peligeet . Th 23D @ $ WIAD. C3 73 6 yo 3B) 36 SkCaf DI Pe 7 lay tt I jo thal ug7 so + 3 448 o- /} i en Ss )) — 1S / Yq 17 SEP 9 1955 LUM b relbica> trans asarth ox ble 920° 22 f 7 pegat 295. Fer prbiremen Lopate Anrnerihe XE Hy, W302 #2, 137 Sg bth, ¢ remomtetc ae — pyri SEP 8 1955 Lesfow selon link ire poorwy HV A. D9 agers. f ages I i Moye] ryedn Ty bane 4C TT it la dat /