title: Techniques of celloidin removal from temporal bone sections.
abstract: OBJECTIVE
  We sought to determine whether the technique of celloidin removal influences the results of immunostaining in celloidin-embedded cochleae.
  
  
  METHODS
  We compared four protocols of celloidin removal, including those using clove oil, acetone, ether-alcohol, and methanol saturated with sodium hydroxide. By optimally fixing our tissue (perfused mice), and keeping constant the fixative type (formalin plus acetic acid), fixation time (25 hours), and decalcification time (ethylenediaminetetraacetic acid for 7 days), we determined whether the technique of celloidin removal influenced the immunostaining results. Six antibodies were used with each removal method: prostaglandin D synthase, sodium, potassium adenosine triphosphatase (Na+,K(+)-ATPase), aquaporin 1, connective tissue growth factor, tubulin, and 200 kd neurofilament.
  
  
  RESULTS
  Clove oil, acetone, and ether-alcohol resulted in incomplete removal of the celloidin, thereby negatively affecting the results of immunostaining. The methanol-sodium hydroxide method was effective in completely removing the celloidin; it produced the cleanest and most reproducible immunostaining for all six antibodies.
  
  
  CONCLUSIONS
  Freshly prepared methanol saturated with sodium hydroxide and diluted 1:2 with methanol was the best solvent for removing celloidin from mouse temporal bone sections, resulting in consistent and reproducible immunostaining with the six antibodies tested.
authors: Department of Otolaryngology, Massachusetts Eye and Ear Infirmary, 243 Charles St, Boston, MA 02114, USA.
authors: Jennifer T
authors: JT
authors: O'Malley
authors: Barbara J
authors: BJ
authors: Burgess
authors: Diane D
authors: DD
authors: Jones
authors: Joe C
authors: JC
authors: Adams
authors: Saumil N
authors: SN
authors: Merchant
mesh_terms: D000096:Acetone / Q000494:pharmacology; D000818:Animals; D045783:Clove Oil / Q000494:pharmacology; D003051:Cochlea / Q000187:drug effects* / Q000378:metabolism* / Q000473:pathology; D003101:Collodion / Q000494:pharmacology*; D000431:Ethanol / Q000494:pharmacology; D004986:Ether / Q000494:pharmacology; D007150:Immunohistochemistry; D000432:Methanol / Q000494:pharmacology; D051379:Mice; D008808:Mice, Inbred CBA; D012997:Solvents / Q000494:pharmacology*; D014014:Tissue Adhesives / Q000494:pharmacology*; D046509:Tissue Culture Techniques; D016610:Tissue Embedding / Q000379:methods*
chemical_list: D045783:Clove Oil; D012997:Solvents; D014014:Tissue Adhesives; D004986:Ether; D000096:Acetone; D000431:Ethanol; D003101:Collodion; D000432:Methanol