Вы находитесь на странице: 1из 7

Connective tissue

Connective tissue fills the spaces between organs

and tissues, and provides structural and metabolic
support for other tissues and organs.
Connective tissue is made up of cells ,fibers and
extracellular matrix
Cells: Fibroblast cell, Adiposite, Hematopoietic stem
cells, Mesenchymal cells, Osteoblasts and Neuroglail
Fibers: Collagen fibers , Elastic fibers , Reticular
Matrix: matrix proteins ,Cell adhesion proteins,
Proteoglycans, water, ions.
Types of connective tissues:
1- C.T. proper : loose (It contains some cells called
'plasma cells', finer elastin fibres and thicker collagen
fibres), dense (fibroblasts)
2- Skeletal C.T.
smooth, cardic)

bone, cartilage,


3- Vascular C.T. (RBCs, WBCs, Platelets)

4- Neural C.T. (Neuroglail cells )
Functions of connective tissues
1.Bind and support the tissues.
2.Connect each organ to other.
3.Transport the nutrition.
4.Helps in the movement of the body tissues.
5.Storage of water, fat and salts.


6.Protects against trauma and pathogenic organisms.

7.Repair the damage tissues by process of fibrosis
Collagenic fibers
It is the main component of connective tissue.
Collagen: is a group of naturally occurring protein.
The fibroblast is the most common cell which creates
Collagen compose of elongated fibrils.
mostly found in fibrous tissue such as tendon,
ligament and skin, and is also abundant in cornea,
cartilage, bone, blood vessels, the gut, and
intervertebral disc.
Histochemical Methods for collagen
collagen fibers are very eosinophilic with
microscope and H&E stain.


The best stain use in differentiating collagen from

other fibers is Trichrome stain which include Massons
Trichrome and VanGieson stain.

H&E stain
Factors affecting trichrome staining
1. Tissue permeability and dye molecular size:

The dye with small molecular weight will penetrate

and stain any of the three tissue type (RBCs, muscle,
medium size dye molecules will penetrate collagen
and muscle. It will not stain RBCs.
The larger size dye will penetrate only collagen.
A larger dye molecule can penetrate the same
element, the smaller molecule will be replaced by it.
2. PH: in order to achieve adequate or even staining
with trichrome stain, dyes should be prepared at
low PH (1.5 to 3).
3. Heat: increase the rate of staining and influence
penetration by larger dye molecules
4. Fixation: the routine formaldehyde fixative is not
optimal for trichrome stain.
zinkers solution, formal mercury or Bouins fixative
give more satisfactory reaction.

Massons trichrome
three dyes are employed selectively staining
section are first stained with an acid dye such as
Biebrich Scarlet
All acidophilic tissue elements such as cytoplasm,
muscles and collagen will bind with acid dyes.
The section are then treated with phosphotungestic
and phosphomolydic acid
the phosphotungestic and phosphomolydic acid
cause the Biebrich Scarlet to diffuse out the collagen
but not the cytoplasm, , muscle, erythrocytes
finally a dye of large particle size such as Aniline blue
is used, that is able to enter collagen.
Tissue Preparation for Trichrome staining
Fixative: Bouin's , zenker`s solution, formal-mercury
are the most satisfactory fixatives for trichrome technique.
Sectioning : Cut paraffin sections 4-5 microns. Frozen
section can be used.
Positive control: Skin, lung, stomach, intestine.
Technical consideration

Decreased red staining usually indicates that the

staining solution has aged or been overused and
should be discarded.
An iron hematoxylin solution is used for nuclear stain
in the trichrome procedure because iron hematoxlyin
more resistant than aluminum hematoxylin to
weigert iron hematoxylin should be prepared fresh.


dark red to black

Cytoplasm, muscle, erythrocytes



VanGieson stain
consider as a primary connective tissue stain to
demonstrate collagen fiber specifically.
used as counter stain in many techniques e.g.

Verhoeff for elastic fiber

Fouchet for bile

Van Kossa for calcium

In strongly acid solution, collagen is selectively
stained by acid fuchsin.
Saturated picric acid provides the acidic pH
necessary and also act as stain of muscle and
The low pH is very important as selective staining of
collagen will not occur at high pH levels.
The addition of o.25ml of Hcl to 100ml of Van Gieson
will sharpen the differentiation between collagen and
Technical consideration
Dehydration should be rapidly in alcohol as Van
Gieson stain will remove if dehydration is prolonged.
An iron hematoxylin solution is used for nuclear stain
in the trichrome procedure because iron hematoxlyin
more resistant than aluminum hematoxylin to
weigert iron hematoxylin should be prepared fresh.

blue black



Muscle & cytoplasm:


Clinical application
Most commonly they are used to demonstrate
To differentiate between collagen and smooth muscle
in tumors, and the increase of collagen in diseases
such as cirrhosis.
Trichrome will also aid in identifying normal
structures, such as connective tissue capsules of
organs, the lamina propria of gastrointestinal tract,
and the bronchovascular structures in lung