Larvae also feature a rich behavioural repertoire which allows them to respond to variations in environmental key factors such as light, hydrostatic pressure, tidal currents, temperature, salinity, and food concentration (reviews ). Larvae develop by successive moults through a species-dependent number of larval instars and, in the absence of parental care, are adapted to developing in the plankton concerning aspects such as movement, nutrition, and sensing (reviews ). Many decapod crustaceans display a complex life cycle which includes pelagic larvae and in many subgroups benthic juvenile-adult stages so that these organisms undergo a distinct niche shift during the transition from larvae to the juvenile phase (reviews ). Larval development of decapod crustaceans embraces a wealth of fascinating ontogenetic processes that are well worth analysing.

Whereas crustacean embryos are popular study objects of developmental biologists (recent reviews, e.g., ), analyses on organogenesis in crustacean larvae somewhat lag behind (see Table 1 in companion paper ) although, for many decades, brachyuran larvae have served as distinguished models in the field of Ecological Developmental Biology (reviews ). For each method, we report our personal experience and give estimations of the method’s research possibilities, the effort needed, costs and provide an outlook for future directions of research.

The methods cover reflected-light-based methods, autofluorescence-based imaging, scanning electron microscopy, usage of specific fluorescence markers, classical histology (paraffin, semithin and ultrathin sectioning combined with light and electron microscopy), X-ray microscopy (♜T), immunohistochemistry and usage of in vivo markers.

Here, we review various classical and more modern imaging techniques suitable for analyses of eidonomy, anatomy, and morphogenetic changes within decapod larval development, and protocols including many tips and tricks for successful research are provided. In a companion paper, we have already described the techniques for culturing larvae in the laboratory and dissecting and chemically fixing their tissues for histological analyses. This publication is the second part of our survey of methods to study organogenesis in decapod crustacean larvae. For the developmental biologist, decapod crustaceans provide the fascinating opportunity to analyse how the adult organism unfolds from organ Anlagen compressed into a miniature larva in the sub-millimetre range. Cells and tissues form the bewildering diversity of crustacean larval organ systems which are necessary for these organisms to autonomously survive in the plankton.

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