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FIGURE 13.5
ImmunoEM using ultrathin sections. (A) Lowicryl HM20 section. 3T3-L1 adipocytes
treated with isoproterenol are quick-frozen, freeze-substituted in 0.2% uranyl acetate and 5%
H 2 O in acetone, and embedded in Lowicryl HM20 resin. LDs are seen as electron-lucent
areas. Inset: An LD of a medium electron density is labeled for PLIN1a. Reproduced in a
modified form from Ariotti et al. (2012) with permission. (B) The Tokuyasu method. HepG2
cells are fixed in 3% formaldehyde in 0.1 M phosphate buffer for 60 min. Ultrathin
sections are labeled by anti-PLIN2 antibody. The LD cores are lost from the section leaving
vacant areas. Colloidal gold labels are observed in the rim of the vacancy (arrows).
(C) The VIS2FIX H method. Ultrathin cryosection of quick-frozen cells is adhered to EM grids
and treated with 0.05% osmium tetroxide. The LD core is seen in a medium electron
density, indicating preservation of lipid esters. Reproduced from Karreman et al. (2011) with
permission.
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