Worksheet 1 Individual Task

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Student name : Dinie Aqilah Ahmad Fariz

Student number : 0330846
Date of submission : 21/9/2017

What do you understand by the term tissue fixative?

Tissue fixative is a fluid that preserves cells in their natural state so that they may be
identified and examined. It is a process by which the cells or tissues are fixed in chemical and
partly physical state so that they can withstand subsequent treatment with various reagents,
with minimal distortion of morphology and no decomposition. The purpose of fixation is to
preserve tissues permanently in as life-like a state as possible.

Describe the procedure of routine Haematoxylin & Eosin staining for tissue sections, and
explain the purpose of each steps involved.

Purpose Reagents
Deparaffinization- removal of paraffin Xylene I
wax. Xylene II
-Hydration step- introduction of water
into the tissue section.
Absolute alcohol
-To remove xylene since it is not miscible
with aqueous solutions.
To rehydrate tissue and prepare it so that
95% alcohol
it can enter purely aqeous solutions.
Rinse off any remaining clearants and
Running water
previous solutions.
-Basic stain that stains acidic structures
(nucleus, ribosomes, rER) purple. Haematoxylin
-Nuclear staining
To remove and rinse off excess dye. Slides Running water
Acid alcohol
are immersed in acidic alcohol for
differentiation purposes and afterwards Running water
the process is halted by a water rinse.
-Nuclei will be stained purplish colour of Blueing (slow running water)
the acid dye. Shifting colour range to blue
provides a much better contrast to the
usual pinkish-red counterstains.
-To remove residual alkali that can
impede eosin staining and cause weak
and uneven staining.
-An acidic dye that stains basic structures
(most cytoplasmic proteins, connective
tissue and muscle fibers) orange-red-
Eosin
pink.
-Used as a counterstain, to contrast the
cytoplasm with the stained nuclei.
Rinse off excess eosin stain. Running water
-Dehydration, to remove any remaining 70% alcohol
80% alcohol
water.
90% alcohol
-To decolorize (weakening the colour) 95% alcohol
highly concentrated eosin solution which Absolute alcohol I
Absolute alcohol II
might cause a blurring of the distinction
between nuclear and non-nuclear Absolute alcohol III
elements.
Xylene I
Used as clearing agents.
Xylene II
-Used as a transition between aqueous
and non-aqueous in order to produce a
clear slide that allows light to pass well
under the microscope (due to its Xylene III
refractive index).
-Activate the adhesive on cover-slippers
using cover-slip tape.
To physically protect the specimen; the
mounting medium bonds specimen, slide
and cover-slip together with a clear
DPX (mounting media)
durable film. The medium is important for
the image formation as it affects the
specimen's rendition.