COMPARATIVE STUDY OF EUCALYPTUS (CITRODORA) OIL AND XYLENE ASCLEARING AND DEWAXING AGENTS IN HISTOLOGICAL AND IMMUNOHISTOCHEMICAL TISSUE PROCESSING AND STAINING
COMPARATIVE STUDY OF EUCALYPTUS (CITRODORA) OIL AND XYLENE ASCLEARING AND DEWAXING AGENTS IN HISTOLOGICAL AND IMMUNOHISTOCHEMICAL TISSUE PROCESSING AND STAINING
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Date
2021-04
Authors
AKPULU, Stephen Peter
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Abstract
Clearing agents are among the most noxious and hazardous chemicals found in histology
laboratories. Xylene has probably been the most commonly used chemical in the histology
laboratory despite its hazards. There have been several laboratory tests to replace xylene as
clearing agent but most of these commercially available substitutes in some cases are less
effective, more expensive,slightly biodegradable and are not readily available. The present study
was aimed at evaluating the clearing and dewaxing effect of Eucalyptus(citrodora oil), and
Xylene during histological and histochemical tissue processing and staining of some selected
organs of adults Wistar rats. Citrodora oil was used alongside xylene to clear livers, kidney,
brain, and testicular tissues of adult Wistar rats. The Citrodora oil was extracted from the leaves
of eucalyptus plants by hydro distillation method. The physical properties, the phytochemical,
Gas Chromatography-Mass Spectroscopy (GC-MS) and Nuclear Magnetic Resonancestudies of
the oil were carried out using standard procedures. The tissues were also histologically processed
using standard method. The resultsshowed that Citrodora oil was pale yellow in color. The
densities of the Citrodora oil and xylene were0.80g/ml and 0.84 g/ml, respectively. The viscosity
of the Citrodora oil was 0.34cp and that forxylene was 0.45cp. The flash point of Citrodora was
at 350C, while that of xylene, was at 390C. The refractive indexes ofCitrodora oil and xylene
were 1.46 and 1.50,respectively. The GC-MS results of Citrodora oil shows that the predominant
constituent in Citrodora oil iscitronellal with relative abundance of 42.20% followed by β-
Citronellol (21.71%). Xylene contains 94% of o-xylene, 97% of p-xylene, 98% of m-xylene and
98%p-xylene. The nuclear magnetic resonance showed that all the chemical shifts are positive to
the reference and are made up of mainly triglycerides. Histological, histochemical and
immunohistochemical observation showed that the section quality, the cytoplasmic and nuclear
staining qualities appeared to give good contrast in Citrodora oil when compared with xylene.
There was no statisticallysignificantdfferences(p≥0.05)in the section quality and staining ability
of liver, kidney, brain and testicular tissues of Wistar rats clearedin Citrodora oil and Xylene
compared. This present study concludes that Citrodora oil can clear and dewax Wistar rat tissues
during tissue and staining processes for histological, histochemical and immunohistochemical
tissue processing and staining as xylene.
Description
A DISSERTATION SUBMITTED TO THE SCHOOL OF
POSTGRADUATE STUDIES, AHMADU BELLO UNIVERSITY, ZARIA,
IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE
AWARD OF DOCTOR OF PHILOSOPHY IN HUMAN ANATOMY
DEPARTMENT OF HUMAN ANATOMY,
FACULTY OF BASIC MEDICAL SCIENCES,
COLLEGE OF MEDICAL SCIENCES,
AHMADU BELLO UNIVERSITY,
ZARIA, NIGERIA
Keywords
COMPARATIVE STUDY,, EUCALYPTUS,, (CITRODORA),, OIL,, XYLENE ASCLEARING,, DEWAXING AGENTS,, HISTOLOGICAL,, IMMUNOHISTOCHEMICAL TISSUE PROCESSING,, STAINING.