cell measurements

1m = 102 cm (centimetres) = 103 mm –

(millimetres) = 106 μm (micrometres) = 109 nm
(nanometres) = 1010 Angstroms (Å)
• cell measurements : Units of measurement in
microns at the light microscope level At the
nanometer level, the electron microscope
 histological preparations
• (1) Anesthesia: some methods of anesthesia
• -10% alcohol is recommended for freshwater animals
• Magnesium chloride is used with marine animals
• In the case of large-sized mammals such as dogs and cats,
the animal is anesthetized with either or chloroform, then
the animal is killed during it is anesthetic
• In the case of small mammals, the animal is killed directly
or dislocation between the cervical vertebrae
• In the case of amphibians, chloroform is anesthetized, or
the animal is killed by hitting the back of the head on the
edge of a table or by pithing.
• In the case of lizards, the animal's head is cut off with
strong scissors
• (2) separation of the sample: The dissection is
done in a physiological solution that reduces the
imbalance that occurs in the nature of the
chemical activity of the organ .
• Among the most famous physiological solutions:
Ringer's solution and saline solution .
• The organ must be removed from the animal's
body and washed in physiological solution to
remove the blood attached to it, because the
blood hinders the fixation process.
• (3) fixation of samples: The organ removed from
the animal’s body is exposed to the
decomposition of its cells and tissues by the
action of bacteria or by the action of enzymes
within the tissues. Therefore, it must be placed in
one of the chemical solutions called fixatives,
which work on: fixed cells and tissues in a state
closer to the living state .
• There are many types of fixatives, including bouin
and formalin
• (4) post -fixation Treatment : To remove the
excess of some components of the fixative
inside the sample, otherwise we got a bad
staining of the sections
• Example: The excess of formalin is removed by
washing the sample in running water for 12
hours
• (5) Dehydration: Most of the fixatives contain water, so it is
necessary to remove the water from the sample before embedding
it in wax. The sample is dehydrated with an increasing series of
ethyl alcohol (70%-80%-95%-two changes of absolute alcohol).
• (6) clearing: It is an intermediate step between the process of
removing water with alcohol and the process of embedding in wax,
since alcohol does not dissolve in wax. The most common material
used in embedding is xylol.
• (7) infiltration: It takes place in the oven by placing four glass
containers, the first containing a mixture of wax and clearing agent,
and the remaining three containers containing molten wax. The
purpose of this process is for the wax to completely permeate the
sample so that it replaces the clearing agent.
• (8) Embedding: A special space is prepared outside the
oven, in which the molten wax is poured, then the
samples are transferred with forceps from a wax
container 3 located in the oven to that space.
• (9) Trimming the wax mold and fixing it on the
microtome: Trimming is the removal of excess wax
around the sample with a sharp blade or scalpel.
• (10) section cutting: by microtome
• (11) affixation of the sections: The paraffin sections
are glued by transferring them with a brush to the
surface of a hot glass slide with a drop of distilled
water on it.
• (12) Dewaxing: Removing the wax surrounding and permeating the
segments using xylol (two changes)
• (13) Hydration: with a descending chain of alcohol (100%-95%-80%-
70%), then distilled water.
• (14) staining
• (15) differentiation : Removing the excess dye from the section.
• The differentiation solution differs from one dye to another
• (16) Dehydration: with an increasing series of alcohol (70%-80%-
95%-100%-100%)
• (17) clearing: by xylol (two changes)
• (18) Mounting (Covering the samples): by the cover glass, using a
mounting media such as Canada balsam