Research paper
Intracellular staining of neurons with nickel-lysine

https://doi.org/10.1016/0165-0270(87)90050-1Get rights and content

Abstract

A new method of intracellularly staining neurons is described. Nickel-lysine (NL) can be used for both intracellular injection by pressure or iontophoresis and retrograde labelling (axonal backfilling). Once introduced into neurons, NL is reacted with dithiooximide dissolved in dimethyl sulfoxide (DMSO) to produce a blue-black precipitate. Small diameter processes are easily detected. For pressure injections, mixing NL with carboxyfluorescein provides a simple way to gauge how much dye has been injected, in that the latter is readily visible when illuminated with blue light. NL appears to move within neurons by axonal transport. Staining over long distances can be obtained in 12–24 h. NL does not appear to cross electrotonic synapses and remains confined to the neurons into which it has been injected. NL staining is simple, flexible and inexpensive. It has the additional advantage that it is compatible with other staining techniques.

References (47)

  • A.R. Light et al.

    Horseradish peroxidase: an improvement in intracellular staining of single, electrophysiologically characterized neurons

    Exp. Neurol.

    (1976)
  • M.-M. Mesulam et al.

    Transganglionic and anterograde transport of horseradish peroxidase across dorsal root ganglia: a tetramethylbenzidine method for tracing central sensory connections of muscles and peripheral nerves

    Neuroscience

    (1979)
  • R.R. Notvest et al.

    Anatomical organization of neurons descending from the supraesophageal ganglion of the lobster

    Brain Res.

    (1981)
  • L.C. Schmued et al.

    Selective neuronal uptake of 4-acetamido-4′-isothiocyanostilbene-2.2′-disulfonic acid (SITS) and other related substituted stilbene in vivo: a fluorescent whole cell staining technique

    Brain Res.

    (1985)
  • L.C. Schmued et al.

    SITS: a covalently bound fluorescent retrograde tracer does not appear to be taken up by fibers of passage

    Brain Res.

    (1982)
  • L.C. Schmued et al.

    Fluoro-Gold: A new fluorescent retrograde axonal tracer with numerous unique properties

    Brain Res.

    (1986)
  • W.W. Stewart

    Functional connections between cells as revealed by dye-coupling with a highly fluorescent naphthalimide tracer

    Cell

    (1978)
  • N.M. Tyrer et al.

    The intensification of cobalt-filled neurone profiles using a modification of Timm's sulphide-silver method

    Brain Res.

    (1974)
  • J.A. Winer

    A review of the status of the horseradish peroxidase method in neuroanatomy

    Biobehav. Rev.

    (1977)
  • W. Winlow et al.

    The morphology of identified neurons in the abdominal ganglion of Aplysia californica

    Brain Res.

    (1976)
  • B. Zipser

    Horseradish peroxidase nerve backfilling in leech

    Brain Res.

    (1980)
  • M. Antal et al.

    Distal dendrites of frog motor neurons: a computer-aided electron microscopic study of cobalt-filled cells

    J. Neurocytol.

    (1986)
  • S. Cullheim et al.

    Combined light and electron microscopic tracing of neurons, including axons and synaptic terminals, after intracellular injection of horseradish peroxidase

    Neurosci. Lett.

    (1976)
  • Cited by (28)

    • Distribution of serotonergic neurons in the eyestalk and brain of the crab, Cancer antennarius

      1990, Comparative Biochemistry and Physiology. Part C, Comparative
    View all citing articles on Scopus
    View full text