Cuticle mounting

Whole-mount Preparation for Staining NMJs of 15-22-hour-old Embryos. This procedure is similar to the method for staining 0-15-hour-old embryos, but it adds a sonication step to allow penetration of the reagents through the cuticle. This protocol is based on a similar protocol described earlier by Patel (1994). 

MOUNTING OF CUTICULAR PREPARATIONS, 602 Mountants, 602 Slides and Coverslip, 602 PREPARATION OF LARVAL CUTICLES, 602 Unhatched First-instar Larval Cuticles, 602 Cuticles of Feeding Larvae, 606... 

Protocol 36.3 Rapid Mounting op Adult Structures in FIoyer s Medium, 612 Protocol 36.4 Preparation and Mounting op Aduet Structures in Canada Balsam, 613 VIEWING CUTICLE PREPARATIONS, 614... 

Hoyer s medium is a useful mountant for both larval and adult cuticles. The medium rapidly digests soft tissues, leaving the cuticle cleared for observation. In addition, samples can be transferred directly from water to Hoyer s medium. However, specimens mounted in Hoyer s medium will degrade over time. For example, the fine denticles on the larval dorsum are best observed soon after mounting they begin to fade after 1 week, and they may disappear completely after several months. More robust features, such as the ventral denticle belts, will persist for a longer period of time. The preparation of Hoyer s medium is described in Protocol 36.1. 

We distinguish between techniques for mounting larval cuticles of first-instar larvae that have not hatched from the egg (Protocol 36.1) from techniques for mounting larvae that have hatched and begun feeding (Protocol 36.2). Protocol 36.1 is for sampling all e s laid by one or more females, which can be particularly useful, for example, when a mutation produces embryos that cannot hatch from the egg. 

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