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PubMedDate not supplied

An improved fixation method for guinea pig cochlear tissues.

The influence of different fixation methods and of various primary fixatives on the ultrastructural preservation of guinea pig cochlear tissues was investigated. No differences in fixation quality were observed between cochleas fixed by intravascular perfusion and cochleas fixed by intralabyrinthine perfusion. Tri-aldehyde primary fixation resulted, in contrast to other formulae investigated, in an excellent, uniform preservation of all cochlear tissues without obvious fixation artefacts. The influence of OSO4/K4Ru(CN)6- and OSO4/K4Fe(CN)6 postfixation was also tested. Cochlear tissues postfixed with OSO4/K4Ru(CN)6 or OSO4/K4Fe(CN)6 exhibited more cellular detail (e.g., membrane- and glycogen contrast) as compared to tissues postfixed with OSO4 alone. Tri-aldehyde primary fixation followed by OSO4/K4Ru(CN)6- or OSO4/K4Fe(CN)6 postfixation therefore is recommended as a multipurpose procedure for optimal preservation of labyrinthine tissues.

Animals↗
PubMed2001

Knee flexion contractures: soft tissue correction with monolateral external fixation.

We assessed the efficacy of progressive soft tissue distraction using monolateral external fixation in the management of severe knee flexion contractures. We prospectively evaluated 10 knee deformities in seven pediatric patients. After gradual distraction using the modified Orthofix Limb Reconstruction System (LRS), most recent functional status and knee range of motion were determined. This treatment was applied to 10 extremities in seven patients, ranging in age from 2 to 16 years. Diagnoses included arthrogryposis (4), sickle cell disease (1), previous sepsis (1), and congenital pterygium (1). Average preoperative flexion contracture was 80.5 degrees. Each patient achieved full extension. There was one recurrence, despite bracing, which was managed with replacement of the fixator and soft tissue procedures. Management of knee flexion contractures using a monolateral fixator appears to be a viable alternative to extensive release or femoral osteotomy. Long-term follow-up will be essential to assess the overall risk of recurrence and complications.

Adolescent↗
PubMed2006

In vivo experimentation with simian herpesviruses: assessment of biosafety and molecular contamination.

In vivo studies with highly pathogenic viruses prompt concerns regarding the persistence of infectious virus in pathology specimens. Although formalin fixation of tissues may inactivate infectious virus, fixation may also degrade viral nucleic acid and antigens, thereby limiting detection of virus in tissues by polymerase chain reaction (PCR) amplification or immunohistochemistry (IHC). We sought to: 1) assess the rate of inactivation of infectious virus in tissue specimens during formalin fixation, 2) assess IHC recognition of viral antigens and PCR detection of viral DNA after long-term (14 d) formalin fixation, and 3) investigate microtome contamination by DNA carry-over to subsequently sectioned tissues. Infectious baboon herpesvirus HVP2 could be recovered from fresh tissues of infected mice but not those fixed in formalin for >/=24 h. The intensity of IHC staining of viral antigen was unaffected by the duration of formalin fixation. PCR detection of viral DNA was negatively impacted by formalin fixation and/or heat inherent to paraffin processing; however, amplification of very short DNA sequences using real-time PCR was not affected. Lastly, microtome contamination by viral DNA was demonstrated by PCR screening of uninoculated control tissues that were sectioned after sectioning infected tissues. In summary, infectious virus is inactivated after only 24 h of formalin fixation whereas IHC staining remains sensitive in tissues fixed for up to 14 d. Formalin fixation does degrade DNA, but viral DNA can be detected by PCR amplification of very short DNA sequences. In addition, viral DNA can contaminate a microtome knife such that subsequently sectioned uninoculated control tissues exhibit false positive PCR amplification.

Animals↗
PubMed1978

Microwave fixation versus formalin fixation of surgical and autopsy tissue.

Microwave irradiation of surgical and autopsy tissue penetrates, fixes, and hardens the tissue almost immediately (the fluid media used in the microwave consisted of saline, ten percent phosphate buffered formalin, and distilled water). Tissue sections from a representative sample of organs were tested. Comparable sections were simultaneously fixed in a phosphate buffered ten percent formalin bath in a vaccum oven as a control. Hematoxylin and eosin were used to stain the sections. Results equal to and superior to the control method were obtained. Saline microwave fixation was superior to formalin microwave fixation. Tissues placed in Zenker's solution and fixed in standard microwave oven (for approximately one minute) yielded results at least equal to conventional Zenker fixation (approximately two hours). No tissue hardening resulted from Zenker microwave fixation. A unique time versus temperature graph (microwave heating curve) reduces individual variation with this technique.

Autopsy↗