Slide Culture

Sterile glass Petri plates Filter paper 

U-shaped sterile glass rod to fit inside dish Alternatively, use Q-tips to support slide Sterile microscope slide and glass coverslip Filter paper 

Prepare glass Petri plate to include filter paper, glass support (or Q-tips) microscope slide and cover slip and autoclave. 

Prepare and autoclave agar. Pour into separate sterile Petri plate and allow to solidify. 

Using an alcohol and flame-sterilized spatula, cut the agar into a grid network of suitable size to fit within the dimensions of the coverslip. 

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Yamanaka, T. and Sagara, N. (1990). Development of basidia and basidiospores from slide-cultured mycelia in Lyophyllum tylicolor (Agaricales). Mycol. Res. 94, 847-850. 

Fig. 6. Slide culture. A definitive diagnosis is usually made by distinctive microscopic features, such as the typical spiral hyphae of T. mentagrophytes var. interdigitale.

The ontogeny of conidiophores and conidia was followed using material collected from the upper portions of infected tillers and from observations of the fungus growing on yeast peptone agar in slide cultures. The cultures had been established from single conidia which had been streaked onto the surface of water agar. The slide cultures were incubated in the dark at 25 2°C for 5 days, then examined at hourly intervals. The development of individual conidiophores and conidia could be followed for more than 20 h. 

Tieman, S., Koch, A. and White, D. (1996). Gliding motility in slide cultures of Myxococcus xanthus in stable and steep chemical gradients. J. Bacterial. 178, 3480-5485. 

Glass depression slide with two concavities for chromatin dispersal (VWR Scientific Products, Micro slide, culture, two depression, cat. 48333-002). 

For accurate estimates with yeast populations of low viability the slide culture technique is the preferred method [3,11]. A suitably diluted suspension of yeast is mixed with just molten wort gelatin and applied to a microscope slide or counting chamber. A coverslip is lowered into position and sealed... 

If cold medium in petri dishes is streaked, rather than mixing the inoculum with the warm medium before pouring into the dishes, the colonies can be more easily removed for further tests. A scheme for selective media and confirmatory tests is given in Table 21.1. Results are given in Table 21,4 for bacterial numbers in pitching yeasts. When lactic acid bacteria are suspected, the plates are held in an atmosphere of carbon dioxide because these bacteria often fail to grow unless the gas is present. Selective media may also be used for slide culture [85, 86]. Microcolonies develop from the individual bacterial cells and may be detected in a few hours by microscopical examination -long before colonies could be seen by the naked eye on the medium in a petri dish. 

The formation of a pseudomycelium is occassionally of diagnostic value, although several species of wine yeast are capable of this type of growth. Formation may be demonstrated by the use of slide cultures (see Slide Culture for demonstration of mycelium pseudomycelium. See 3.5.5). 

For observation of molds, slide cultures are typically employed. The procedure described below permits visualization of the structure directly or after staining with minimal mechanical damage to the organism. 

Incubate at room temperature until growth is noted. The slide culture may be removed for microscopic examination during this period. To reduce the likelihood of contamination, examination should be minimized and restricted to high-dry or lower magnifications. 

The presence or absence of mycelium or pseudomycelium is demonstrated by growing the fungi as a slide culture (Section 12.6). However, microscopic evaluation requires removal of the slide culture from the Petri plate, thereby increasing the potential for microbial contamination. To minimize contamination, the number of examinations should be limited such that results can be obtained after only one to two observations. 

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