Microwave-stimulated formaldehyde fixation of experimental renal biopsy tissues: computerised morphometric analysis of distortion artefacts.

Looi, Lai Meng, and Loh, Kum Chee, (2005) Microwave-stimulated formaldehyde fixation of experimental renal biopsy tissues: computerised morphometric analysis of distortion artefacts. Malaysian Journal of Pathology, 27 (1). pp. 23-27. ISSN 0126-8635

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Affiliations

University of Malaya, Medical Centre, Dept. of Pathology
University of Malaya, School of Medical Laboratory Technology

Abstract

Although microwave irradiation has been used in the histopathology laboratory for several years, there has been minimal published experimental data on its effects on the technical and staining quality of histological sections. Furthermore, it has not been clear whether the advantages gained in reduction of fixation and staining duration has been at the expense of increasing architectural distortion to the tissues. We report here our experience with computerised morphometric analysis to investigate glomerular artifacts caused by microwave-stimulated fixation of renal tissues. 39 rat and 33 human autopsy kidney samples were subjected to (1) fixation in neutral buffered formaldehyde (control), (2) microwave-stimulated fixation followed by neutral buffered formaldehyde, and (3)
neutral buffered formaldehyde followed by microwave irradiation. In addition, the effect of postfixation in 70% ethanol was also investigated. Microwave irradiation was delivered through a dedicated laboratory microwave oven at 80% power and at 55°C for 3 minutes. The different fixation methods were compared with regards to shrinkage (distortion) to glomerular structures (glomeruli
and Bowman’s spaces) on H&E sections, as determined by morphometric image analysis using a temporary assembled-system consisting of a trinocular microscope, a digital video camera and personal computer. A FlashPoint VGA 3.3 film-grabber card was used to capture images for morphometric analysis by using a Scion Image program. Morphometric analysis of glomerular structures showed that microwaves caused more shrinkage to the area bounded by the Bowman’s capsule than the glomerulus proper, but post-fixation with ethanol reduced this shrinkage. These findings have implications on the logistics of tissue preparation of renal biopsies in clinical practice.

Item Type:

Journal

Keywords:

Renal biopsy, microwave-fixation, artefact, computerized morphometry

Subjects:

R Medicine, Dentistry, Pharmacy, Nursing

ID Code:

1835