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  • Transmission electron microscopy of thin sections of Drosophila: preparation of embryos using n-heptane and glutaraldehyde.

Transmission electron microscopy of thin sections of Drosophila: preparation of embryos using n-heptane and glutaraldehyde.

Cold Spring Harbor protocols (2012-10-03)
Kent L McDonald, David J Sharp, Wayne Rickoll
ABSTRACT

This protocol describes the simultaneous permeabilization of Drosophila embryos with n-heptane and initial fixation with glutaraldehyde. Even though the vitelline membrane around the embryo is chemically permeabilized, it must be manually removed to achieve infiltration with embedding resins. Once the embryo is embedded, it can be sectioned for transmission electron microscopy (TEM). This procedure can produce excellent preservation for ultrastructural analysis, and is useful for situations where optimal preservation (e.g., by high-pressure freezing) is not required or is not feasible.

MATERIALS
Product Number
Brand
Product Description

Supelco
Heptane, analytical standard
Sigma-Aldrich
Heptane, anhydrous, 99%
Sigma-Aldrich
Heptane, biotech. grade, ≥99%
Sigma-Aldrich
Heptane, HPLC Plus, for HPLC, GC, and residue analysis, 99%
Sigma-Aldrich
Heptane, puriss. p.a., reag. Ph. Eur., ≥99% n-heptane basis (GC)
Sigma-Aldrich
Heptane, ReagentPlus®, 99%
Sigma-Aldrich
Heptane, suitable for HPLC, ≥99%
Sigma-Aldrich
Heptane, suitable for HPLC, ≥96%