Tissue Processing and Common Fixatives Overview
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TISSUE
PROCESSING
1.
Fixation
2.
Dehydration
3.
Clearing
4.
Impregnation
5.
Embedding
and
blocking
6.
Section
cutting
7.
Routine
staining
undefinedCOMMONLY
USED
FIXATIVES
⚫
Formalin
–
MC
–
routine
⚫
Glutaraldehyde
–
electron
microscopy
⚫
Picric
acid(Bouin’s
solution)
–
renal
&
testicular
tissue
⚫
Alcohol(Carnoy’s
fixative)
–
cytologic
smears,
endometrial
sampling
⚫
Osmium
tetraoxide
–
CNS
tissues
&
electron
microscopy
Pathophysiology
of
Endocrine
System
Dr.
Athmar mohamed
Pituitary
disorders
Causes
:
tumor
hemorrhage
trauma
irradiation
Pituitary
disorders
G
r
o
w
t
h
h
o
r
m
o
n
e
d
e
f
i
c
i
e
n
c
y
children
-
short
stature
(
p
i
t
u
i
t
a
r
y
d
w
a
r
f
i
s
m
)
,
n
o
r
m
a
l
i
n
t
e
l
l
i
g
e
n
c
e
,
o
b
e
s
i
t
y
adults
-
central
obesity,
reduced
muscle
mass
and
exercise
capacity.
(pituitary
dwarfism),
Pituitary
disorders
E
x
c
e
s
s
o
f
G
H
i
n
c
h
i
l
d
h
o
o
d
P
i
t
u
i
t
a
r
y
g
i
g
a
n
t
i
s
m
growth
velocity
proportional
enlargement
of
skeleton
and
inner
organs
enlargement
of
the
peripheral
nerves
delayed
puberty
and
hypogonadism
Pituitary
gigantism
Pituitary
disorders
E
x
c
e
s
s
o
f
G
H
i
n
a
d
u
l
t
s
A
c
r
o
m
e
g
a
l
y
reason
–somatotrope
adenoma
hyperplasia
and
hypertrophy
of
soft
tissues
size
of
hands,
feet,
nose,
ears
cartilaginous
proliferation
of
the
larynx
coarsening
of
the
facial
features.
enlarged
tongue
and
increase
of
inner
organs
thick
and
dark
skin.
degenerative
arthritis.
Acromegaly
Thyroid
disorders
.
G
o
i
t
e
r
-
s
i
z
e
o
f
t
h
e
t
h
y
r
o
i
d
g
l
a
n
d
(not
related
of
TH
level)
Complications
of
goiter:
difficulty
in
swallowing,
distention
of
the
veins
of
the
neck
and
upper
extremities,
edema
of
the
eyelids
and
conjunctiva,
syncope
with
coughing.
Goit
e
r
Thyroid
disorders
C
o
n
g
e
n
i
t
a
l
h
y
p
o
t
h
y
r
o
i
d
i
s
m
-
c
r
e
t
i
n
i
s
m
C
a
u
s
e
s
congenital
absence
of
the
thyroid
gland
abnormal
biosynthesis
of
thyroid
hormone
deficient
TSH
secretion.
Clinics:
mental
retardation,
impaired
growth.
Thyroid
disorders
A
c
q
u
i
r
e
d
h
y
p
o
t
h
y
r
o
i
d
i
s
m
–
m
y
x
e
d
e
m
a
accumulation
of
a
mucopolysaccharide
substance
in
the
connective
tissues.
Causes
destruction
or
dysfunction
of
the
thyroid
gland,
impaired
pituitary
function
hypothalamic
dysfunction.
Disorders
of
gall
bladder
By
Dr.
Athmar mohamed
•
2
main
types
of
gall
stones
:
–
Cholesterol
stones
–
Pigment
stones
•
In
the
West,
about
90%
are
cholesterol
stones.
•
Pigment
gall
stone
is
predominant
in
non-
Western
population
–
associated
with
bacterial
infection
of
biliary
tree
and
parasitic
infestations.
Cholesterol
stone
•
Content
:
Crystalline
cholesterol
monohydrate
is
predominant
Pigment
stone
•
Bilirubin
calcium
salt
is
predominant
•
Pigment
stones
–
Black
pigment
–
,
hemolytic
anemia
(hemoglobinopathy,
red
cell
disorders)
–
Brown
pigment
–
Asian
patients
(infection)
Pathology
Cholesterol
stones
:
–
Gross
:
pale
yellow,
ovoid,
firm,
single
to
multiple
with
faceted
surfaces
–
Mostly
radiolucent,
20%
is
radio
opaque
due
to
the
presence
of
calcium
carbonate
content.
:
Pigment
stones
–
Black
stone
(in
sterile
gall
bladder
bile)-
small
size,
fragile
to
touch,
numerous,
50-
70%
are
radioopaque
–
Brown
stone
(in
infected
intrahepatic
or
extrahepatic
ducts)-
single
to
a
few,
soft,
greasy,
soaplike
consistency
due
to
presence
of
retained
fatty
acids
released
by
bacterial
phospholipases
on
biliary
lecithins,
radiolucent.
–
Stone
content
:
calcium
salts
of
unconjugated
bilirubin,
lesser
amounts
of
other
calcium
salts,
mucin
glycoproteins
and
cholesterol.
Summary
:
•
Complications
:
–
Inflammation
of
gall
bladder
(cholecystitis)
–
Empyema
–
Perforation,
–
Fistulas
•
Complications-
cont
–
Inflammation
of
biliary
tree
(cholangitis)
–
Obstructive
cholestasis
–
Pancreatitis
–
Erode
adjacent
bowel
and
cause
intestinal
obstruction
(gallstone
ileus)
Summary:
Pathology
of
acute
cholecystitis
•
Gross
:
–
Enlarged,
tense,
edematous,
red
or
violaceous
colour
(subserosal
haemorrhage)
–
Fibrinous
/fibrinopurulent
exudate
covering
the
serosa
–
+-
stones
obstructing
the
neck
or
cystic
duct
–
Lumen
contains
blood
and
pus
(empyema)
–
Green
black
necrotic
•
Microscopic
:
–
acute
inflammation
in
the
wall
–
mucosal
ulceration.
–
May
be
associated
with
abscess
formation
or
gangrenous
necrosis.
•
Clinical
features
:
–
recurrent
attacks
of
epigastric
or
right
upper
quadrant
pain
–
Nausea,
vomiting
and
intolerance
to
fatty
foods.
•
Pathology:
–
Gross
:
•
smooth
and
glistening
to
dull
serosa
(subserosal
fibrosis)
•
thickened
wall,
opaque
gray-
white
appearance
•
Uncomplicated
cases,
lumen
contains
clear,
green,
mucoid
bile
and
stones
with
normal
mucosa
•
Microscopic
:
–
Reactive
proliferation
of
mucosa
–
Inflammation
(lymphocytes,
plasma
cells,
and
macrophages
in
the
mucosa
and
in
the
subserosal
fibrous
tissue).
May
be
minimal.
–
Prominent
outpouching
of
the
mucosal
epithelium
through
the
wall
(Rokitansky
Aschoff
sinuses)
–
Marked
subepithelial
and
subserosal
fibrosis
–
+-
Superimposed
acute
inflammation
–
+-
Extensive
calcification
within
the
wall
→porcelain
gall
bladder
→increase
risk
of
cancer
•
Xanthogranulomatous
cholecystitis:
massively
thickened
wall
with
shrunken,
nodular,
chronically
inflamed
with
foci
of
necrosis
and
haemorrhage.
•
Hydrops
of
the
gall
bladder
:
atrophic,
chronically
obstructed
gall
bladder
containing
only
clear
secretion
Complications
of
cholecystitis
•
Bacterial
superinfection
with
cholangitis
or
sepsis
•
Gall
bladder
perforation
and
local
abscess
formation
•
Gall
bladder
rupture
with
diffuse
peritonitis
•
Biliary
enteric
(cholecystenteric)
fistula,
with
drainage
of
bile
into
adjacent
organs,
entry
of
air
and
bacteria
into
biliary
tree
and
potentially
gallstone-
induced
intestinal
obstruction
(ileus)
•
Aggravating
of
preexisting
medical
illness,
with
cardiac,
pulmonary,
renal
or
liver
decompensation
•
Porcelain
gall
bladder
with
increased
risk
of
cancer
•
Treatment
:
Cholecystectomy
In tissue processing, the steps include fixation, dehydration, clearing, impregnation, embedding, section cutting, and routine staining. Common fixatives like Formalin, Glutaraldehyde, and Picric acid are used for different tissues. Understand the pathophysiology of the endocrine system and pituitary disorders such as growth hormone deficiency and excess of growth hormone in children and adults. Explore thyroid disorders like goiter and their complications.
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Presentation Transcript
TISSUE PROCESSING Fixation 1. Dehydration 2. Clearing 3. Impregnation 4. Embedding and blocking 5. Section cutting 6. Routine staining 7.
COMMONLY USED FIXATIVES Formalin MC routine Glutaraldehyde electron microscopy Picric acid(Bouin s solution) renal & testicular tissue Alcohol(Carnoy s fixative) cytologic smears, endometrial sampling Osmium tetraoxide CNS tissues & electron microscopy
Pathophysiology of Endocrine System Dr.Athmar mohamed
Pituitary disorders Causes : tumor hemorrhage trauma irradiation
Pituitary disorders Growth hormone deficiency children - short stature (pituitary dwarfism), normal intelligence, obesity adults - central obesity, reduced muscle mass and exercise capacity. (pituitary dwarfism),
Pituitary disorders Excess of GH in childhood Pituitary gigantism growth velocity proportional enlargement of skeleton and inner organs enlargement of the peripheral nerves delayed puberty and hypogonadism Pituitary gigantism
Pituitary disorders Excess of GH in adults Acromegaly reason somatotrope adenoma hyperplasia and hypertrophy of soft tissues size of hands, feet, nose, ears cartilaginous proliferation of the larynx coarsening of the facial features. enlarged tongue and increase of inner organs thick and dark skin. degenerative arthritis. Acromegaly
Thyroid disorders Goiter Goiter - size of the thyroid gland (not related of TH level) Complications of goiter: difficulty in swallowing, distention of the veins of the neck and upper extremities, edema of the eyelids and conjunctiva, syncope with coughing. .
Thyroid disorders Congenital hypothyroidism - cretinism Causes congenital absence of the thyroid gland abnormal biosynthesis of thyroid hormone deficient TSH secretion. Clinics: mental retardation, impaired growth.
Thyroid disorders Acquired hypothyroidism myxedema accumulation of a mucopolysaccharide substance in the connective tissues. Causes destruction or dysfunction of the thyroid gland, impaired pituitary function hypothalamic dysfunction.
Disorders of gall bladder By Dr.Athmar mohamed
2 main types of gall stones : Cholesterol stones Pigment stones In the West, about 90% are cholesterol stones. Pigment gall stone is predominant in non- Western population associated with bacterial infection of biliary tree and parasitic infestations.
Pigment stone Bilirubin calcium salt is predominant Pigment stones Black pigment , hemolytic anemia (hemoglobinopathy, red cell disorders) Brown pigment Asian patients (infection) Cholesterol stone Content : Crystalline cholesterol monohydrate is predominant
Pathology Pigment stones Cholesterol stones : : Black stone (in sterile gall bladder bile)- small size, fragile to touch, numerous, 50-70% are radioopaque Brown stone (in infected intrahepatic or extrahepatic ducts)- single to a few, soft, greasy, soaplike consistency due to presence of retained fatty acids released by bacterial phospholipases on biliary lecithins, radiolucent. Stone content : calcium salts of unconjugated bilirubin, lesser amounts of other calcium salts, mucin glycoproteins and cholesterol. Gross : pale yellow, ovoid, firm, single to multiple with faceted surfaces Mostly radiolucent, 20% is radio opaque due to the presence of calcium carbonate content.
Complications : Inflammation of gall bladder (cholecystitis) Empyema Perforation, Fistulas
Complications-cont Inflammation of biliary tree (cholangitis) Obstructive cholestasis Pancreatitis Erode adjacent bowel and cause intestinal obstruction (gallstone ileus)
Pathology of acute cholecystitis Gross : Enlarged, tense, edematous, red or violaceous colour (subserosal haemorrhage) Fibrinous /fibrinopurulent exudate covering the serosa +- stones obstructing the neck or cystic duct Lumen contains blood and pus (empyema) Green black necrotic Microscopic : acute inflammation in the wall mucosal ulceration. May be associated with abscess formation or gangrenous necrosis.
Clinical features : recurrent attacks of epigastric or right upper quadrant pain Nausea, vomiting and intolerance to fatty foods. Pathology: Gross : smooth and glistening to dull serosa (subserosal fibrosis) thickened wall, opaque gray-white appearance Uncomplicated cases, lumen contains clear, green, mucoid bile and stones with normal mucosa
Microscopic : Reactive proliferation of mucosa Inflammation (lymphocytes, plasma cells, and macrophages in the mucosa and in the subserosal fibrous tissue). May be minimal.
Prominent outpouching of the mucosal epithelium through the wall (Rokitansky Aschoff sinuses) Marked subepithelial and subserosal fibrosis +-Superimposed acute inflammation +-Extensive calcification within the wall porcelain gall bladder increase risk of cancer
Xanthogranulomatous cholecystitis: massively thickened wall with shrunken, nodular, chronically inflamed with foci of necrosis and haemorrhage. Hydrops of the gall bladder : atrophic, chronically obstructed gall bladder containing only clear secretion
Complications of cholecystitis Bacterial superinfection with cholangitis or sepsis Gall bladder perforation and local abscess formation Gall bladder rupture with diffuse peritonitis Biliary enteric (cholecystenteric) fistula, with drainage of bile into adjacent organs, entry of air and bacteria into biliary tree and potentially gallstone-induced intestinal obstruction (ileus) Aggravating of preexisting medical illness, with cardiac, pulmonary, renal or liver decompensation Porcelain gall bladder with increased risk of cancer
