Label-free Chemical Imaging of Fungal Spore Walls by Raman Microscopy and Multivariate Curve Resolution Analysis.
Fungal cell walls are medically important since they represent a drug target site for antifungal medication. So far there is no method to directly visualize structurally similar cell wall components such as α-glucan, β-glucan and mannan with high specificity, especially in a label-free manner. In this study, we have developed a Raman spectroscopy based molecular imaging method and combined multivariate curve resolution analysis to enable detection and visualization of multiple polysaccharide components simultaneously at the single cell level. Our results show that vegetative cell and ascus walls are made up of both α- and β-glucans while spore wall is exclusively made of α-glucan. Co-localization studies reveal the absence of mannans in ascus wall but are distributed primarily in spores. Such detailed picture is believed to further enhance our understanding of the dynamic spore wall architecture, eventually leading to advancements in drug discovery and development in the near future.
Raman Center for Medical and Biological Applications, Shimane University, Matsue 690-8504, Japan.
Hemanth
H
Noothalapati
Raman Center for Medical and Biological Applications, Shimane University, Matsue 690-8504, Japan.
Takahiro
T
Sasaki
Raman Center for Medical and Biological Applications, Shimane University, Matsue 690-8504, Japan.|Faculty of Life and Environmental Science, Shimane University, Matsue 690-8504, Japan.
Tomohiro
T
Kaino
Raman Center for Medical and Biological Applications, Shimane University, Matsue 690-8504, Japan.|Faculty of Life and Environmental Science, Shimane University, Matsue 690-8504, Japan.
Makoto
M
Kawamukai
Consolidated Research Institute for Advanced Science and Medical Care, Waseda University, Tokyo 162-0041, Japan.
Masahiro
M
Ando
Consolidated Research Institute for Advanced Science and Medical Care, Waseda University, Tokyo 162-0041, Japan.|Institute of Molecular Science and Department of Applied Chemistry, National Chiao Tung University, Hsinchu 30010, Taiwan.
Hiro-O
HO
Hamaguchi
Raman Center for Medical and Biological Applications, Shimane University, Matsue 690-8504, Japan.|Faculty of Life and Environmental Science, Shimane University, Matsue 690-8504, Japan.
Tatsuyuki
T
Yamamoto
D002473:Cell Wall / Q000737:chemistry*; D005936:Glucans / Q000032:analysis; D008351:Mannans / Q000032:analysis; D015999:Multivariate Analysis; D000073758:Nonlinear Optical Microscopy; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D013172:Spores, Fungal / Q000737:chemistry*
D005936:Glucans; D008351:Mannans