Clawed Frog Nuclear Envelope Protocol

Application Note for Leica EM CPD300 - Life Science Research

October 06, 2016

Species: Clawed frog (Xenopus laevis)
Critical point drying of nuclear pores from clawed frog oocytes with subsequent chromium coating and SEM analysis

Sample Holder: Silicon chips containing the samples were placed into the filter discs and porous pots holder.

Isolated nuclear envelopes were prepared from Xenopus oocytes as described by Goldberg MW, Fiserova J. (2010) Immunogold labelling for scanning electron microscopy. Methods Mol Biol. 657:297-313.

Fixation and Dehydration:

2% Glutaraldehyde, 0.2% Tannic acid, 0.1M Hepes buffer1x10 min.
Distilled water2x1 min.
0.1% aqueous OsO41x10 min.
Distilled water3x10 min.
Ethanol series: 50%, 70%, 95%1x2 min.
Ethanol series: 100%2x2 min.

Leica EM CPD300 auto program:

Chromium: 1.5 nm

Fig.1: Nuclear pores from clawed frog oocytes
Fig.2: Nuclear pores from clawed frog oocytes