The generalized isolation chip (giChip) is a device created with reference to the previously reported iChip (ref.) microbial culture device. It consists of a central panel with a through-hole for fixed culturing microorganisms and two supporting side panels for surrounding and fixing the culture holes of central panel with a semipermeable membrane. The microorganisms suspended in molten agar are dispensed into the through holes in the central panel, then sandwiched between the semipermeable membrane and side panels, and finally assembled by screwing them up using the outermost circular through hole.The giChip is then placed in soil from which the sample originated.Nutrients can be taken in from the external environment, while preventing microorganisms from entering the culture pore from the outside, which allows the microorganisms to be isolated and cultured in an environment similar to a place in which they were collected.
panel size : 60 mm × 60 mm × 5 mm (each panel)pore size : 2.5 mm in diameter (Through-hole for culture), 4 mm (for fixation).material : polycarbonate
The ordinary culture methods.
After removing dead leaves and other debris covering the soil surface, a soil—as a source of microorganisms at a depth of approximately 3–10 cm below the soil surface—is collected with a sterilized tool, immediately transferred to a sterilized container, sealed, and used on the day of collection. First, the soil is suspended in sterile distilled water and filtered through a filter paper (Advantech, Filter size 6 µm) to obtain a microbial solution. Next, the microbial solution is diluted in molten SMS media (0.125 g casein, 0.1 g potato starch, 1 g casamino acids, and 20 g bacto-agar in 1 liter of water). Then, an average of one cell per culture pore is fixed in the culture holes of central panel. The device is then assembled and incubated in the soil from which the microorganisms are collected at 25°C for 1~4 weeks under shading in the laboratory. SMS agar in each culture pore is subsequently dropped onto Mueller–Hinton agar in 10 cm culture dish and cultured at 25°C for 1 week ~ one month to obtain colonies.
panel size : 60 mm × 60 mm × 5 mm (each panel)pore size : 2.5 mm in diameter (Through-hole for culture), 4 mm (for fixation).material : polycarbonate
The ordinary culture methods.
After removing dead leaves and other debris covering the soil surface, a soil—as a source of microorganisms at a depth of approximately 3–10 cm below the soil surface—is collected with a sterilized tool, immediately transferred to a sterilized container, sealed, and used on the day of collection. First, the soil is suspended in sterile distilled water and filtered through a filter paper (Advantech, Filter size 6 µm) to obtain a microbial solution. Next, the microbial solution is diluted in molten SMS media (0.125 g casein, 0.1 g potato starch, 1 g casamino acids, and 20 g bacto-agar in 1 liter of water). Then, an average of one cell per culture pore is fixed in the culture holes of central panel. The device is then assembled and incubated in the soil from which the microorganisms are collected at 25°C for 1~4 weeks under shading in the laboratory. SMS agar in each culture pore is subsequently dropped onto Mueller–Hinton agar in 10 cm culture dish and cultured at 25°C for 1 week ~ one month to obtain colonies.