Technical report: laser microdissection of cochlear structures from celloidin embedded human temporal bone tissues and detection of the mitochondrial DNA common deletion using real time PCR.
Laser microdissection (LMD) has been used to isolate groups of cells and single cells from numerous tissues. In this study, we describe a technique for isolating cochlear structures and individual spiral ganglion cells from archival celloidin embedded human temporal bone sections. The specimens isolated are suitable for quantifying the mitochondrial DNA (mtDNA) common deletion (CD) within these tissues using a real time polymerase chain reaction (PCR) assay. The results presented in this manuscript demonstrate the feasibility of using this LMD technique to study the accumulation of mtDNA deletions in diseases of the ear. To our knowledge, this approach to analyzing archival human temporal bone tissues has not been previously reported.
Markaryan
Adam
A
Section of Otolaryngology - Head and Neck Surgery, Department of Surgery, University of Chicago, 5841 S. Maryland Avenue, MC 1035, Chicago, IL 60637, United States. amarkary@surgery.bsd.uchicago.edu
Nelson
Erik G
EG
Tretiakova
Maria
M
Hinojosa
Raul
R
D000293:Adolescent; D000368:Aged; D001842:Bone and Bones / Q000378:metabolism*; D003101:Collodion / Q000737:chemistry*; D017931:DNA Primers / Q000737:chemistry; D004272:DNA, Mitochondrial / Q000378:metabolism; D017353:Gene Deletion; D034381:Hearing Loss / Q000378:metabolism*; D006801:Humans; D007834:Lasers; D042282:Microdissection; D008954:Models, Biological; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013701:Temporal Bone / Q000378:metabolism*
D017931:DNA Primers; D004272:DNA, Mitochondrial; D003101:Collodion