Application of cell-surface engineering for visualization of yeast in bread dough: development of a fluorescent bio-imaging technique in the mixing process of dough.
Cell-surface engineering (Ueda et al., 2000) has been applied to develop a novel technique to visualize yeast in bread dough. Enhanced green fluorescent protein (EGFP) was bonded to the surface of yeast cells, and 0.5% EGFP yeasts were mixed into the dough samples at four different mixing stages. The samples were placed on a cryostat at -30 degrees C and sliced at 10 microm. The sliced samples were observed at an excitation wavelength of 480 nm and a fluorescent wavelength of 520 nm. The results indicated that the combination of the EGFP-displayed yeasts, rapid freezing, and cryo-sectioning made it possible to visualize 2-D distribution of yeast in bread dough to the extent that the EGFP yeasts could be clearly distinguished from the auto-fluorescent background of bread dough.
Maeda
Tatsuro
T
Research Center for Basic Science, Nisshin Seifun Group, Inc, Fujimino, Saitama, Japan. maedatat@mail.ni-net.co.jp
Shiraga
Seizaburo
S
Araki
Tetsuya
T
Ueda
Mitsuyoshi
M
Yamada
Masaharu
M
Takeya
Koji
K
Sagara
Yasuyuki
Y
D001709:Biotechnology / Q000379:methods; D001939:Bread / Q000382:microbiology*; D005456:Fluorescent Dyes / Q000378:metabolism*; D005511:Food Handling; D005615:Freezing; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D008163:Luminescent Measurements / Q000379:methods*; D008856:Microscopy, Fluorescence; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000302:isolation & purification* / Q000378:metabolism; D013997:Time Factors
D005456:Fluorescent Dyes; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins