Abstract
Single cell cultivation of bacteria used to require a highly advanced technique and much experience. Recently, however, technical difficulties have been overcome to a large extent by the appearance of the de Fonbrune pneumatic micromanipulator attached with a specially designed microf orge and the phase contrast microscope. Our previous paper (Katsuyama et al., 1957) reported single cell cultivation of E. coli with such apparatus for the study of streptomycin resistance. The cultivation was conducted by so-called “agar film method” originally developed by Kyo (1935) and Minomo (1942) . The technique was described in detail in that paper (Katsuyama et al., 1957) . Beside of it, the authors have been endeavoring also to devise a single cell culture method for tubercle bacilli. Because, despite of our repeated trials, the original “agar film method” never succeeded in making a single cell of the tubercle bacillus develop to form a visible colony in concave space of a depression slide. Such cultivation environment was considered unsuitable for tubercle bacilli which require a prolonged period for visible colony formation. To the best of our knowledge, there has never been a report of satisfactory method for single cell culture of tubercle bacilli as yet. However, the present authors have finally reached a new device applicable to the above purpose. The present paper is principally describing the technical aspects together with examples of its application to drug sensitivity test.
