Ultrathin cryosectioning

Cryomicrotomy and cryoultranucrotomy are sectioning methods performed at low temperatures to produce thin or ultrathin sections, respectively, of pol)m ers too soft for room 

Early results with cryomicrotomes were described by Cobbold and Mendelson [80]. Polyurethane elastomer, a blend of crystalline and noncrystalline polymers, showed spherulitic textures after sectioning at about -70°C. Injection molded polypropylene (PP) was also sectioned at about -70°C, while polytetrafluoroethylene (PTFE) was sectioned at much lower temperatures. The authors concluded that the technique, though difficult, had potential. Extruded styrene-butadiene-styrene (SBS) copolymer was prepared by cryosectioning with a diamond knife in liquid air at —85 to —115°C, followed by osmium tetroxide vapor staining for one hour [81]. This method revealed the alternating sequence of the polystyrene and polybutadiene lamellae. Odell et al. [82] prepared extruded triblock copolymer by first chemically hardening the polybutadiene, with osmium tetroxide, followed by cryoultramicrotomy to produce 30 nm thick sections which showed fine structure details. Parallel polystyrene rods were observed in the SBS copolymer. Ultramicrotomy and selective staining with osmium tetroxide was also used in the preparation of a binary blend of PP and thermoplastic rubber [83]. 

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Nielsen, M.H., Bastholm, L., Chatterjee, S., Koga, J., and Norrild, B. (1989) Simultaneous triple-immunogold staining of virus and host cell antigens with monoclonal antibodies of virus and host cell antigens in ultrathin cryosections. Elistochemistry 92, 89-93. 

Frey B, Brunner I, Walther P, Scheidegger C, Zierold K. Element localization in ultrathin cryosections of high-pressure frozen ectomycorrhizal spruce roots. Plant Cell Environ 1997 20 929-937. 

Slot, J. W. and Geuze, H. J. (1984) Gold markers for single and double immuno-labeling of ultrathin cryosections, in Immunolabelling for Electron Microscopy (Polak, J. M. and Vamdell, I. M., eds.), Elsevier, New York, pp. 129-142. 

Chicoine, L., and Webster, P. 1998. Effect of microwave irradiation on antibody labeling efficiency when applied to ultrathin cryosections through fixed biological material. Microsc. Res. Tech. 42 24-32. 

Takizawa, T., Suzuki, K., and Robinson, J. M. (1998) Correlative microscopy using FluoroNanogold on ultrathin cryosections proof of principle. J. Histochem. Cytochem. 46, 1097-1102. 

J., Hyttinen, M.M., Jurvelin, J.S., and Helminen, HJ., Reference sample method reduces the error caused by variable cryosection thickness in Fourier transform infrared imaging, Appl. Spectrosc. 58, 137-140, 2004 Takizawa, T. and Robinson, J.M., Thin is better Ultrathin cryosection immunocytochemistry, J. Nippon Med. Sch. 71, 306-307, 2004. 

Fig. 14. Localization of PhoE-LacZ hybrid proteins in Escherichia coli by means of cell fractionation studies (A) showing clearly the presence of the hybrid protein in the total cellular proteins of induced cells (lane e), the cell envelopes of induced cells (lane f) and the Triton X-100 insoluble protein fraction of the cell envelopes (lane h) and with the immunogold technique on ultrathin cryosections an accumulation of hybrid protein in the cytoplasm is clearly shown (B). Bar = 0.2 jam. From [165],...

Takizawa T, Robinson JM (2003) Ultrathin cryosections an important tool for immimo-fluorescence and correlative microscopy. J Histochem Cytochem 51 707-714... 

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