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The Direct Experimental Observation of Cells in Phenomic Lag



2 is dry the drop ia rapidly absorbed XXI microuco!Sc observation May be Gegun almet immediately. The mechanical stqo is rs.xbfied to accomod~te Q petri dish, the corner of the grid is located under low trti then Ngh JTower and, as the grid is scanned, each cell or configur&tir~n (singlet, doublet, triplet etc.) is dmwn on a xper facsimile of the grid. .,t timed Sntttrvtla tit-3 grid is scanned dnd the number of cells in each configunrticn is alrain recorded. In- termittent RlfcroscopPc obsarv&tion may be made of the open -)Lato for hii hours without vibrible interference from airborne contamtints, The tendency of growth to beco!.~e confluent interferes tith longor ~~sriods of observation. Two plating media have been emloyed. Cne, a ,r:lucoso nutrient aga &P- lows both resliratiun deficient (asr) and resri_riAtion sufficiant (gL13> csUs lx form colonies. T. e other, a lactate nutrient aqar allous on& .;CKH cells to form coIktxaies (5). When a respiration deficient clonal fsolaio of strain l4940 (61, grating in exponential phase %n glucor;e nutrient broth was trans- ferred ti glucose g&d als;rtes, essentially all cell8 or cnnfigurtitions de- veloped to form W&X&38. A low frequency (less than 1%) failed to ;lroduce a aw,le bud and wme considered dead colle. PJht3n aL3.puot.s cf the sa:nt, Cal- ture were studied on lact&e grid pljrtes, not a sin@ new bdd W&R formed in more th;;m loo0 Cunffgurationa exa&ned. To test whether the aor cells M died on ffre lachte agar a doughnut shaped ring of filter p;por irapregntrted with rplucoae wa:; placed on the agar surface surrounding the grid, dter some l&g essentially all configurations budded on the s-ltitos to which glucose had been ad,ied. The aer cells wore themfom still viable. blthough previous obeervatlons (5) had indfcated that aor cells were incapsibla of for&g macro- colonies on lactitc ago, the observation cf the co:r~~:lete failure to form I-W buds on the U~

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