MicroTopic ‘To study the bacterial cell cycle-for instance. to determine ‘where in the oycle DNA synthesis occurs-one needs a popula tion of cells all atthe same stage. With such a population, cells can be exposed. for instance. to radioactive thymidine to mea- sure the rate of DNA synthesis. A number of various diferent ‘ways to synchronize cultures were tried. but these depended on ‘manipulations that had the potential to disturb the cel cycle ‘and create misleading results. Thus the invention, by Charles Helmstetter and Stephen Cooper, of the simple and elegant “baby machine.” was a significant advance. They began by fiter ingacutureot growing E col through a very ine pore iter used normally to sterilize heat sensitive media (see Section 3.19) ‘was known that bacteria not only cannot passthrough the fine pores of such fiter but also actually sicko its surface (because interactions between the LPS ofthe bacterialoutermemirane and the surface of the fiter) So Helmstetter and Cooper simply ‘umed the fiter ver and tickled fresh medium through the fier ‘An exponentaly roving ‘ule of Ecole iered ‘Syria ot by ‘araton ne trison / (aoe ‘ios alae aod | brates moe “etre wae tried wih baby Eat caectd rom a baby machne ‘hohe average ero es Inewaton me inn) Cities the Le (Figure 98.1),Asthe cols onthe iter divided, they released baby Cellinto the medium flowing by. Thus. cells in the effluent from the fiter would consis of baby cals only. Figure 9.6 shows the ‘population growth of a sample taken from a baby machine ‘larly the population is synchronized, although there is consi ‘erable heterogeneity inthe interdvision times. The baby machine produces baby cells that can be used to stansynctvonized cultures that can be usedto study the cal cycle ‘experimental by for instance. adding radioactive labels atvarious ;pointsin thecal cycle and determining the love's of incorporation into various macromolecules, Alteratiely, an exponentially ‘growing culture can be labeled frst then used to start a baby ‘machine. The amount of radioactivity in cels released from the ‘baby machine at diferent times then allows calculation oftherates ‘of macromolecular synthesis in each sogment ofthe cal cyclo. ‘as this latter approach that was used to work out the details of| ‘overlapping cel cycles in bacteria. as described in Section 9.7 “The iter is then turned cover and fesh medium owed though belie released, FIGURE 98.1. Diagram of the “baby machine.”