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Nabeel-------------------------------Histology------------------------------2nd Stage
Microtechniques (Microscopic preparations):-
The microtechniques for tissue preparation consist from many steps to study the
cellular structure which forming the body of the living human or animals, which
can't see by naked eyes, but can see by using Light microscope or Electronic
microscope.
* There are three types of methods using for microscopic preparations are:
1- The paraffin Technique.
2- The celloidin technique.
3- The freezing technique.
*The paraffin technique:
In this technique using paraffin wax, following are the steps used to work
histological sections and are summarized as follows:
1- Obtaining of the specimen.
2- Fixation.
3- Washing.
4- Dehydration.
5- Clearing.
6- Impregnation or Infiltration.
7- Embedding.
8- Trimming.
9- Sectioning.
10- Mounting.
11- Rehydration.
12- Staining.
13- Cover slipper.
Dr. Nabeel-------------------------------Histology------------------------------2nd Stage
1- Obtaining of the specimen: Possible to obtain of the Biopsy sample from a
human patient during surgery and
animal after killing him directly the
animal is permanent stop and fast
for all biological processes and can
be make the killing of the animal in
several ways (Slaughter - Paralyzed
Animal - Hit the back of the head -
Anesthesia).
2- Fixation: The first step in the preparation of tissue using 10% formalin
solution. The purpose from using formalin to maintaining the tissue and its
contents to the state it was in the body living or near from that and the fixation
process through chemical reactions and interactions physical between the active
groups of the fixative substance and effective groups of chemicals in tissue
(Carbohydrate - protein - fat - enzymes - metal salts-dyes). And the fixation
process made upon stop fragmentation and disintegration process for bacteria and
fungi activity, as well as stop the autolysis process of the tissue by enzymes.
3-
Washing : The sample must be washed before and after fixation by using tap
water current for 24 hours, to remove the remainder of the formalin from the
specimen.
Dr. Nabeel-------------------------------Histology------------------------------2nd Stage
4- Dehydration: Dehydration by using series spiraling
of ethyl alcohol beginning with ( 50%, 60%,70%, 80%,
90%, 90%, 100% and 100% ) for 2 hours for each
concentration, to gradual removal of water from the
tissue to prevent tissue shrinking.
9- Sectioning:-Paraffin block are cut by microtome using metal knife, into thin
sections ~ 6μ.