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animal slide preparation technique

animal slide preparation technique

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Published by: aesha89 on Oct 20, 2009
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02/01/2013

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Animal Slide Preparation Technique
INTRODUCTION :

Histology is the study of the microscopic anatomy of cells and tissues of plants and animals. It is all about the study of structure and function of each tissue types, how those tissues are combined to form the organs and systems of the body, and how those combinations function together. It is performed by examining a thin slice of tissue under a light microscope or electron microscope. The ability to visualize or differentially identify microscopic structures is frequently enhanced through the use of histological stains. Histology is an essential tool of biology and medicine.

Fixation is a chemical process by which biological tissues are preserved from decay, either through autolysis or putrefaction. Fixation terminates any ongoing biochemical reactions, and may also increase the mechanical strength or stability of the treated tissues. Fixation is usually the first stage in a multistep process to prepare a sample of biological material for microscopy or other analysis. Therefore, the choice of fixative and fixation protocol may depend on the additional processing steps and final analyses that are planned.

A microtome is a mechanical instrument used to cut biological specimens into transparent thin sections for microscopic examination. Microtome use steel, glass, or diamond blades depending upon the specimen being sliced and the desired thickness of the sections being cut. Steel blades are used to prepare sections of animal or plant tissues for light microscopy histology. Glass knives are used to slice sections for light microscopy and to slice very thin sections for electron microscopy. Industrial grade diamond knives are used to slice hard materials such as bone, teeth and plant matter for both light microscopy and for electron microscopy. Gem quality diamond knives are used for slicing thin sections for electron microscopy.

There are some precaution when we are using the microtome while doing the slicing.
Microtome use extremely sharp blades. All operations, with the blade, must be done with the

greatest of care. There are some operations where the hands must be brought close to the blade. For these procedures hand protection must be worn for example is cut resistant gloves. Then, the knives need to be cleaned. Disposable knives have an anticorrosion coating that must be removed. Diamond paste must be removed from knives when they are sharpened. The smallest silicon particle can ruin the sections. All dust must be removed. Such careful cleaning must bring the hands close to the blade with the attendant risk.

Biological tissue has little inherent contrast in either the light or electron microscope. Staining is employed to give both contrast to the tissue as well as highlighting particular features of interest. Where the underlying mechanistic chemistry of staining is understood, the term histochemistry is used. Hematoxylin and eosin (H&E) is the most commonly used light microscopical stain in histology and histopathology. Hematoxylin stains nuclei blue, eosin stains the cytoplasm pink. Uranyl acetate and lead citrate are commonly used to impart contrast to tissue in the electron microscope.

MATERIALS:

Mould
Forceps
Slides and cover slides
Microtomes
Beakers
Normal saline solution
70%, 80%, 95% and 100% Ethanol
Bouin\u2019s solution
Toluena solution
Paraffin solution
Xilen 1
Xilen 2

Absolute alcohol + xilen (1:1)
Absolute alcohol
Alcohol90%
Alcohol 80%
Alcohol75 %
Distilled water
Hematoxilin solution
0.5% Hydrochloric solution
0.5% sodium bicarbonate solution
Scott\u2019s solution
Eosin
DPX medium/Balsam Canada

METHODS:
TISSUE PREPARATION:
1. An animal was given to each group.
2. The animal was operated and the organs was taken
a.
Liver
b.
Lung
c.
Intestine
d.
Esophagus
e.
Heart
f.
Ovary / testis
g.
Uterus

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