(A) Ultrathin section of P. tricornutum expressing Pex10-GFP in Epon without antibody labeling. The boxed area is shown in (B) at higher magnification and illustrates two peroxisomes in proximity to the nucleus, the golgi and the plastid. CW, cell wall; G(A) Ultrathin section of P. tricornutum expressing Pex10-GFP in Epon without antibody labeling. The boxed area is shown in (B) at higher magnification and illustrates two peroxisomes in proximity to the nucleus, the golgi and the plastid. CW, cell wall; G

Targeting of peroxisomal matrix proteins in the diatom Phaeodactylum tricornutum

Sample Preparation:
P. tricornutum cells expressing different types of GFP fusion proteins were harvested via centrifugation at 1,500xg and cryoimmobilized by high-pressure freezing on gold carriers (Leica EM PACT2, Leica Microsystems GmbH, Vienna, Austria). Subsequently, the cells were freeze-substituted with acetone in combination with 2% OsO4, 0.25% uranyl acetate and 5% H2O. Freeze substitution was carried out in the automated Leica EM AFS2 unit (Leica Microsystems GmbH, Vienna, Austria) at -90°C for 4 h, -60°C for 8 h, -30°C for 8 h and then held at 0°C for at least 3 h ...

For more details please see:

http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0025316

Courtesy of:
Dr. Kathrin Bolte and Dr. Andreas Klingl
Zellbiologie & Elektronenmikroskopie
AG Prof. Uwe-G. Maier
Philipps Universität Marburg, Marburg
The website of the EM facility is:
http://www.synmikro.com/de/startseite/core-facilities/elektronenmikroskopie.html

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Related Images

(A) Ultrathin section of P. tricornutum expressing Pex10-GFP in Epon without antibody labeling. The boxed area is shown in (B) at higher magnification and illustrates two peroxisomes in proximity to the nucleus, the golgi and the plastid. CW, cell wall; G