Microwave Decalcification of Human Temporal Bones

Cunningham, Calhoun D.; Schulte, Bradley A.; Bianchi, Lynne M.; Weber, Peter; Schmiedt, Barbara N.

doi:10.1097/00005537-200102000-00017

Cited by 42 publications

(39 citation statements)

References 12 publications

(24 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Cunningham et al (2001) described an alternative for IHC of human temporal bones that used a decalcification method with a microwave oven process. A good signal-to-noise ratio of the immunostaining of Na + K + -ATPase in the human organ of Corti, as well as acceptable morphological preservation, was demonstrated.…”

Section: Introductionmentioning

confidence: 99%

Immunohistochemical techniques for the human inner ear

López

Ishiyama

Hosokawa

et al. 2016

Histochem Cell Biol

View full text Add to dashboard Cite

In this review, we provide a description of the recent methods used for immunohistochemical staining of the human inner ear using formalin-fixed frozen, paraffin and celloidin-embedded sections. We also show the application of these immunohistochemical methods in auditory and vestibular endorgans microdissected from the human temporal bone. We compare the advantages and disadvantages of immunohistochemistry (IHC) in the different types of embedding media. IHC in frozen and paraffin-embedded sections yields a robust immunoreactive signal. Both frozen and paraffin sections would be the best alternative in the case where celloidin-embedding technique is not available. IHC in whole endorgans yields excellent results and can be used when desiring to detect regional variations of protein expression in the sensory epithelia. One advantage of microdissection is that the tissue is processed immediately and IHC can be made within 1 week of temporal bone collection. A second advantage of microdissection is the excellent preservation of both morphology and antigenicity. Using celloidin-embedded inner ear sections, we were able to detect several antigens by IHC and immunofluorescence using antigen retrieval methods. These techniques, previously applied only in animal models, allow for the study of numerous important proteins expressed in the human temporal bone potentially opening up a new field for future human inner ear research.

show abstract

Section: Introductionmentioning

confidence: 99%

Immunohistochemical techniques for the human inner ear

López

Ishiyama

Hosokawa

et al. 2016

Histochem Cell Biol

View full text Add to dashboard Cite

show abstract

“…Here, the Sox10 expression data demonstrated the utility of our temporal bone preparation procedure [43], [71] and immunostaining assay using a frozen sectioning process to identify a particular transcription factor with low expression in human postmortem cochlear tissues. Further quantitative studies of lateral wall degeneration associated with candidate regulatory factors (identified from animal models) in human temporal bones will significantly advance our understanding of the causes of presbyacusis and other auditory disorders.…”

Section: Discussionmentioning

confidence: 82%

“…The fixative was perfused gently through the oval window using a blunt-tip, 16-gauge needle covered with appropriately sized tygon tubing. The perfused temporal bones were then immersed in fixative for 48–72 hours for frozen section preparation and 12–22 hours for paraffin section preparation followed by rinsing and decalcification using a microwave protocol as per our previous description [43]. The total time of decalcification was between 3–6 weeks.…”

Section: Methodsmentioning

confidence: 99%

Sox10 Expressing Cells in the Lateral Wall of the Aged Mouse and Human Cochlea

et al. 2014

Self Cite

View full text Add to dashboard Cite

Age-related hearing loss (presbycusis) is a common human disorder, affecting one in three Americans aged 60 and over. Previous studies have shown that presbyacusis is associated with a loss of non-sensory cells in the cochlear lateral wall. Sox10 is a transcription factor crucial to the development and maintenance of neural crest-derived cells including some non-sensory cell types in the cochlea. Mutations of the Sox10 gene are known to cause various combinations of hearing loss and pigmentation defects in humans. This study investigated the potential relationship between Sox10 gene expression and pathological changes in the cochlear lateral wall of aged CBA/CaJ mice and human temporal bones from older donors. Cochlear tissues prepared from young adult (1–3 month-old) and aged (2–2.5 year-old) mice, and human temporal bone donors were examined using quantitative immunohistochemical analysis and transmission electron microscopy. Cells expressing Sox10 were present in the stria vascularis, outer sulcus and spiral prominence in mouse and human cochleas. The Sox10+ cell types included marginal and intermediate cells and outer sulcus cells, including those that border the scala media and those extending into root processes (root cells) in the spiral ligament. Quantitative analysis of immunostaining revealed a significant decrease in the number of Sox10+ marginal cells and outer sulcus cells in aged mice. Electron microscopic evaluation revealed degenerative alterations in the surviving Sox10+ cells in aged mice. Strial marginal cells in human cochleas from donors aged 87 and older showed only weak immunostaining for Sox10. Decreases in Sox10 expression levels and a loss of Sox10+ cells in both mouse and human aged ears suggests an important role of Sox10 in the maintenance of structural and functional integrity of the lateral wall. A loss of Sox10+ cells may also be associated with a decline in the repair capabilities of non-sensory cells in the aged ear.

show abstract

“…4,5,15 However, more recent findings show that microwaving significantly reduces decalcification time, with no obvious adverse effects on the structure and antigenicity. 2,7,10 Microwaving also reduces the time taken for fixation, dehydration, infiltration, clearing, and staining.…”

Section: Decalcificationmentioning

confidence: 99%