Influence of initial fixation protocol on the appearance of primary cilia on the rabbit corneal endothelial cell apical surface as assessed by scanning electron microscopy

Authors

  • Michael J Doughty

    Corresponding author
    1. Department of Vision Sciences, Glasgow-Caledonian University, City Campus, Glasgow, Scotland G4 0BA, UK
      Corresponding author. Tel.: +44-141-331-3393; fax: +44-141-331-3387 M.Doughty@gcal.ac.uk
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Corresponding author. Tel.: +44-141-331-3393; fax: +44-141-331-3387 M.Doughty@gcal.ac.uk

Abstract

The objective was to examine primary cilia at the apical surface of corneal endothelial cells after using different fixatives. Female albino rabbits (2 kg) were euthanised at 15:00 h and the corneas fixed immediately (usually with an isotonic 2% glutaraldehyde—cacodylate fixative) either after dissection, by application fixative at 4 °C, by immersion of the eyeball in fixative at room temperature (RT), or by application of an isotonic or a hypertonic (Karnovsky-type) fixative at RT. Images at 2000× were taken from the central corneal region, and number and length of primary cilia assessed. The length was the same regardless of method (overall average of 1.67±0.70 μm), but the incidence of primary cilia was hypertonic fixative (87% of cells) >cold drop fixation (71%), >whole globe immersion (68%) >dissect then fix methods (67%) >RT drop fixation (34%). The first four methods however yielded cells with unacceptable artefacts (especially distortion). More details should be provided of the primary fixation method used.

Ancillary