A method for efficient observation of intracellular membranes of monolayer culture cells by quick-freeze and freeze-fracture electron microscopy

Journal of Electron Microscopy

Volumn 61, Issue 6, 2012, Pages 441-446

  Fujita, Akikazu ^a,b   Fujimoto, Toyoshi ^a   Ozato Sakurai, Nami ^a   Suzuki, Hitomi ^a  

^a NAGOYA UNIVERSITY GRADUATE SCHOOL OF MEDICINE   (Japan)

^b KAGOSHIMA UNIVERSITY   (Japan)

Author keywords

electron microscopy; freeze fracture; high pressure freezing; lipid; metal contact freezing

Indexed keywords

ARTICLE; CELL LINE; CELL ORGANELLE; FREEZE FRACTURE; HUMAN; METHODOLOGY; TRANSMISSION ELECTRON MICROSCOPY; ULTRASTRUCTURE;

CELL LINE; FREEZE FRACTURING; HUMANS; MICROSCOPY, ELECTRON, TRANSMISSION; ORGANELLES;

ELECTRON MICROSCOPES; ELECTRON MICROSCOPY; FRACTURE; FREEZING;

CULTURE CELLS; FREEZE FRACTURE; FREEZE FRACTURE ELECTRON MICROSCOPY; FREEZE-FRACTURE REPLICA; HIGH-PRESSURE FREEZING; INTRACELLULAR MEMBRANES; LABELINGS; METAL CONTACT FREEZING; METAL CONTACTS; MONOLAYER CULTURE;

LIPIDS;

EID: 84871204420     PISSN: 00220744     EISSN: 14779986     Source Type: Journal    
DOI: 10.1093/jmicro/dfs063     Document Type: Article

References (19)

1. Collins V P, Arborgh B, and Brunk U (1977) A comparison of the effects of three widely used glutaraldehyde fixatives on cellular volume and structure. A TEM, SEM, volumetric and cytochemical study. Acta Pathol. Microbiol. Immunol. Scand. A l85A: 157-168.
2. Lee R M, McKenzie R, Kobayashi K, Garfield R E, Forrest J B, and Daniel E E (1982) Effects of glutaraldehyde fixative osmolarities on smooth muscle cell volume, and osmotic reactivity of the cells after fixation. J. Microsc. 125: 77-88.
3. Johnson T J A (1985) Aldehyde fixatives: quantification of acidproducing reactions. J. Electron Microsc. Techn. 2: 129-138.
4. Schlörmann W, Steiniger F, Richter W, Kaufmann R, Hause G, Lemke C, and Westermann M (2010) The shape of caveolae is omega-like after glutaraldehyde fixation and cup-like after cryofixation. Histochem. Cell Biol. 133: 223-228.
5. Smitherman M L, Lazarow A, and Sorenson R L (1972) The effect of light microscopic fixatives on the retention of glycogen in protein matrices and the particulate state of native glycogen. J. Histochem. Cytochem. 20: 463-471.
6. Jost P, Brooks U J, and Griffith O H (1973) Fluidity of phospholipid bilayers and membranes after exposure to osmium tetroxide and gluteraldehyde. J. Mol. Biol. 76: 313-318.
7. Mayor S, Rothberg K G, and Maxfield F R (1994) Sequestration of GPI-anchored proteins in caveolae triggered by cross-linking. Science 264: 1948-1951.
8. Fujita A, Cheng J, Hirakawa M, Furukawa K, Kusunoki S, and Fujimoto T (2007) Gangliosides GM1 and GM3 in the living cell membrane form clusters susceptible to cholesterol depletion and chilling. Mol. Biol. Cell 18: 2112-2122.
9. Fujita A, Cheng J, and Fujimoto T (2010) Quantitative electron microscopy for the nanoscale analysis of membrane lipid distribution. Nat. Protoc. 5: 661-669.
10. Fujita A, Cheng J, and Fujimoto T (2009) Segregation of GM1 and GM3 clusters in the cell membrane depends on the intact actin cytoskeleton. Biochim. Biophys. Acta. 1791: 388-396.
11. Fujita A, Cheng J, Tauchi-Sato K, Takenawa T, and Fujimoto T (2009) A distinct pool of phosphatidylinositol 4,5-bisphosphate in caveolae revealed by a nanoscale labeling technique. Proc. Natl Acad. Sci. USA 106: 9256-9261.
12. Moor H (1987) Theory and practice of high pressure freezing. In: Steinbrecht R A and Zierold K (eds), Cryotechniques in Biological Electron Microscopy, pp 175-191 (Springer, Berlin).
13. Stauffer T P, Ahn S, and Meyer T (1998) Receptor-induced transient reduction in plasma membrane PtdIns(4,5)P2 concentration monitored in living cells Curr. Biol. 8: 343-346.
14. Bulbarelli A, Sprocati T, Barberi M, Pedrazzini E, and Borgese N (2002) Trafficking of tail-anchored proteins: transport from the endoplasmic reticulum to the plasma membrane and sorting between surface domains in polarised epithelial cells. J. Cell Sci. 115: 1689-1702.
15. Sawaguchi A, Aoyama F, Ide S, Goto Y, and Suganuma T (2008) A new device for high-pressure freezing of cultured cell monolayer using 10-microm-thin stainless discs as both culture plate and specimen carrier. J. Electron Microsc. (Tokyo) 57: 203-206.
16. Heuser J E, Reese T S, Dennis M J, Jan Y, Jan L, and Evans L (1979) Synaptic vesicle exocytosis captured by quick freezing and correlated with quantal transmitter release. J. Cell Biol. 81: 275-300.
17. Sitte H, Edelmann L, and Neumann K (1987) Cryofixation without pretreatment at ambient pressure. In: Steinbrecht R A and Zierold K (eds), Cryotechniques in Biological Electron Microscopy, pp 87-113 (Springer, Berlin).
18. Takahashi T, Hirsh A, Erbe E, and Williams R J (1988) Mechanism of cryoprotection by extracellular polymeric solutes. Biophys. J. 54: 509-518.
19. McDonald K L and Auer M (2006) High-pressure freezing, cellular tomography, and structural cell biology. Biotechniques 41: 137-143.