Kerosene: Contributing agent to xylene as a clearing agent in tissue processing

Abstract

Background:Research methodology in oral and maxillofacial pathology has illimitable potential. The tissue processing involves many steps of which one of the most important step is “Clearing,” which is a process of replacing dehydrant with a substance which is miscible with embedding medium or paraffin wax. Xylene is one of the common clearing agents used in laboratory, but it is also hazardous. The main aim of this study is to substitute conventionally used xylene by a mixture of kerosene and xylene in clearing steps without altering the morphology and staining characteristics of tissue sections. This will also minimize the toxic effects and tend to be more economical.Materials and Methods:One hundred and twenty bits of tissue samples were collected, each randomly separated into 4 groups (A, B, C and D) and kept for routine tissue processing till the step of clearing; during the step of clearing instead of conventional xylene, we used mixture of xylene and kerosene in 4 ratios ([A-K:X – 50:50]; [B-K:X – 70:30]; [C – Ab. Kerosene]; [D – Ab. Xylene – as control]) and observed for the light microscopic study adopting H and E staining, IHC (D2-40), Special stains (periodic acid–Schiff and congo red) procedure. The result was subjected to statistical analysis by using Fisher's exact test.Results:The results obtained from the present study were compared with control group, i.e., D and it was observed that Groups A and B were absolutely cleared without altering the morphology of tissue and cellular details; optimum embedding characteristics and better staining characteristics were also noted, whereas Group C presents poor staining characteristics with reduced cellular details. Embedded tissues in Group C presented with rough, irregular surface and also appeared shrunken.ConclusionCombined mixture of xylene and kerosene as a clearing agent in different ratio, i.e., Group A (K:X – 50:50) and B (K:X – 70:30) can be used without posing any health risk or compromising the cellular integrity.