FIGURE 3: Morphology and immunolabeling of yeast cells embedded in acrylic resins or processed following the Tokuyasu method.

(A) Yeast was cryofixed in propane, freeze-substituted in acetone containing 3% GA and embedded in Lowicryl HM20 at low temperatures. Specific antibodies and protein A were used to localize the COX complex. IB, inclusion body; IBM, inclusion body with membrane; M, mitochondria; N, nucleus. Scale bar, 0.5 µm. This image was originally published in [157] © Springer.

(B) Cells were fixed in GA/PFA, dehydrated with ethanol and embedded in LR White resin. Immunolabeling was directed to cell wall antigens. K, karmellae; M, mitochondria; N, nucleus. This image was originally published in [6] © John Wiley and Sons.

(C) Cells were fixed with 4% PFA and 0.4% GA, treated with sodium metaperiodate, embedded in 12% gelatin and infiltrated with 2.3 M sucrose before being frozen in liquid nitrogen. Atg9 was localized with antibodies and protein A-gold. CW, cell wall; M, mitochondria; PM, plasma membrane; V, vacuole. Scale bar, 0.5 µm. This image was originally published in [159] © Mari et al, 2010.

6. Wright R. (2000). Transmission electron microscopy of yeast. Microsc.Res.Tech. 51(6): 496-510. http://www.ncbi.nlm.nih.gov/pubmed/?term=11169854

157. Binder M, Hartig AT. (1996). Immunogold labeling of yeast cells: an efficient tool for the study of protein targeting and morphological alterations due to overexpression and inactivation of genes. Histochem.Cell Biol. 106(1): 115-130. http://dx.doi.org/10.1007/BF02473206

159. Mari M, Griffith J, Rieter E, Krishnappa L, Klionsky DJ, Reggiori F. (2010). An Atg9-containing compartment that functions in the early steps of autophagosome biogenesis. J.Cell Biol. 190(6): 1005-1022. http://dx.doi.org/10.1083/jcb.200912089