Dictyostelium Cell Fixation: Two Simple Tricks

We share two simple modifications to enhance the fixation and imaging of relatively small, motile, and rounded model cells. These include cell centrifugation and the addition of trace amounts of glutaraldehyde to existing fixation methods. Though they need to be carefully considered in each context, they have been useful to our studies of the spatial relationships of the microtubule cytoskeletal system.

Lavender, a chick melanocyte mutant with defective melanosome translocation: a possible role for 10 nm filaments and microfilaments but not microtubules

Lavender is a mutation of chick neural-crest-derived melanocytes showing dilute feather pigmentation. This defect, previously attributed to a lack of attenuation of dendrites, was found to be due to a defect in melanosome translocation. The mutant phenotype, of melanincongested perikarya and pigmentless dendrites is expressed both in vivo and in vitro. Studies with colcemid and cytochalasin B suggest that the avian melanocyte resembles a dispersing amphibian melanophore in its requirement for microfilaments but not microtubules. Ultrastructural analysis revealed a normal complement of intracellular filaments. Microtubules, however, are scarce. Intermediate (10 nm) filaments surround and are closely associated with intracellular organelles, while microfilaments interconnect all filaments and organelles. While-cell centrifugation at 300 g showed that 10 nm filaments stream behind and appear to attach to mobile membrane-bound organelles including the nucleus, lipid granules and mitochondria, as well as melanosomes. It is suggested that all intracellular filaments, especially microfilaments and intermediate filaments, interconnect forming a network responsible for organelle motility.