Optimization of methylmethacrylate infiltration duration in undecalcified murine bone specimens

Melissa Kacena, Jana K. Halfon, Christiane E. Coady, Tracy Nelson, Nancy W. Troiano

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

To obtain high-quality, undecalcified bone sections, specimens must be properly fixed, dehydrated, cleared, infiltrated, embedded, sectioned, and stained. Here, we examine different infiltration durations for methylmethacrylate-embedded mouse tibiae and lumbar vertebrae. Our data indicate that 24 h of vacuum infiltration allows for good sections in both lumbar vertebrae and tibiae, with minimal microtears. Furthermore, we show that with 24 h of infiltration, excellent results were obtained for all seven histochemical and immunohistochemical stains performed. In general, good results were obtained when specimens were infiltrated for 12-72 h. However, with 120 h of infiltration, the marrow retained methylmethacylate even after the sections were deplasticized. Also, when specimens were infiltrated for 6 h, they were underinfiltrated, resulting in increased numbers of microfractures in trabecular bone spicules. Therefore, in today's busy laboratory setting, in which quickly examining a bone phenotype, performing histomorphometry, or special bone-specific stains is of importance, we recommend that mouse tibiae and lumbar vertebrae be infiltrated under vacuum for 24 h at 4°C with methylmethacrylate before embedding. This technique provides a good balance between section/staining quality and time.

Original languageEnglish (US)
Pages (from-to)21-27
Number of pages7
JournalJournal of Histotechnology
Volume29
Issue number1
StatePublished - Mar 2006
Externally publishedYes

Fingerprint

Methylmethacrylate
Lumbar Vertebrae
Tibia
Vacuum
Bone and Bones
Coloring Agents
Stress Fractures
Bone Marrow
Staining and Labeling
Phenotype

Keywords

  • Antigen preservation
  • Enzyme preservation
  • Histology
  • Methylmethacrylate infiltration technique
  • Morphological preservation
  • Mouse
  • Plastic infiltration

ASJC Scopus subject areas

  • Anatomy
  • Cell Biology

Cite this

Optimization of methylmethacrylate infiltration duration in undecalcified murine bone specimens. / Kacena, Melissa; Halfon, Jana K.; Coady, Christiane E.; Nelson, Tracy; Troiano, Nancy W.

In: Journal of Histotechnology, Vol. 29, No. 1, 03.2006, p. 21-27.

Research output: Contribution to journalArticle

Kacena, M, Halfon, JK, Coady, CE, Nelson, T & Troiano, NW 2006, 'Optimization of methylmethacrylate infiltration duration in undecalcified murine bone specimens', Journal of Histotechnology, vol. 29, no. 1, pp. 21-27.
Kacena, Melissa ; Halfon, Jana K. ; Coady, Christiane E. ; Nelson, Tracy ; Troiano, Nancy W. / Optimization of methylmethacrylate infiltration duration in undecalcified murine bone specimens. In: Journal of Histotechnology. 2006 ; Vol. 29, No. 1. pp. 21-27.
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