 Autopsy Descending

-remove from a dead person 1. Filing

2. Labelling
-24 hours of death or to know the cause of 3. Mounting
death 4. Staining
- for a cause of death / final diagnosing 5. Sectioning
6. Trimming
a. above or below never sideways 7. Embedding
(parents/siblings/offspring/) 8. Impregnation
9. Clearing/de-alcoholization
b. Sideways (spouse)
10. Dehydration
CHARACTERISTICS OF AUTOPSY 11. Decalcification
12. Fixation
-post mortem
 FIXATION
- everything will be remove
-life like structure
- to see for one self
-must be compatible of your stain
- to save lives, to understand more about the
disease/contribute to medical knowledge -Preservation (morphe and chemical)

TYPES OF HOMICIDAL -to prevent post mortem changes

- Infantricide – “infant” Example:

- Fratricide – “Killing ones brother
- Sororicide – “sister” 1. Autolysis – Self distraction and it is
- Patricide – “parents” from its own enzymes. (own cell)
- Uxoricide – “one’s wife” 2. Heterolysis – Enzyme from another or
- Filicide – “one’s child” different cell.
- Regicide – “monarch” 3. Putrefaction – foul odor
- Genocide – “national or ethnic religious 4. Decomposition – Bacteria
people” a. Pseudomonas aeroginusa –
- Fruity odor
 Biopsy 5. Necrosis – Cell death (due to hypoxia
and it would lead to ischemia)
-remove from a living person for treatment or a. Coagulation – can be found in
true diagnosis Myocardial Infarction
b. Causation or caseation- can be
found in MPV.
12 STEPS OF HISTOPATH c. Liquefaction – Abscess
d. Gangrenous – combination of
Ascending coagulation and liquefaction
and can be found in lower
1. Fixation
limbs.
2. Decalcification
3. Dehydration
 To prevent the post mortem we can use
4. Clearing/de-alcoholization
Isopentane in liquid nitrogen to
5. Impregnation
invivofication.
6. Embedding
7. Trimming Mordant – Potassium alum
8. Sectioning
9. Staining -bridge from primary to secondary stain.
10. Mounting
Fixative – antiseptic and bactericidal effect or
11. Labelling
resistant of different changes of reagents.
12. Filing
EFFECTS OF FIXATION
1. Hardening of tissue (coagulation)
2. Decrease risk of nfection
a. Antiseptic
b. Bacterial effect
3. Act as a mordant
a. Increase affinity to the skin
4. Minimize post mortem changes
5. Resistant to different concentration of
reagent
6. Be better or increase of optical
differentiation
OBJECTIVES OR ADVANTAGES OF FIXATIVE
1. Cheap
2. Stable
3. Easily available
4. Easily prepared
5. Strong action
6. Strong penetration
7. It must be safe
8. It must be isotonic
REQUIREMENTS
1. Amount of fixative
a. Normally – 10-20x
i. Osmic acid - <10x
ii. For museum – 30-50x
(Kaisserlings – most
common used for
museum) in normally
used 10% of formalin
b. Thickness – must be 3-5 mm
sample thickness except for
halo organs (lungs, brain,
embryo) at least 1cm cut.
c. Duration
i. 8 hrs. – room temp.
ii. 6 hrs. – room temp +
agitation
iii. 45˚ celsius – is 4.8 – 8
hours (25% -40%) of the
original time.
d. Container – must be wide
mouth
6 FACTORS OF CHANGES IN DURATION
1. Mucus – wash with NSS
2. Blood – wash with NSS
3. Fats – cut thinner
4. Quality of Reagent
5. Urgency of process
6. Skills
7. Temperature (not sure)
TYPES OF FIXATIVE ACCORDING TO THEIR
ACTION AND COMPOSITION
A. Simple fixatives – made up of only one
component substance
1. Aldehydes
a. Formaldehyde
b. Glutaraldehyde
2. Metallic Fixatives
a. Mercuric Chloride
b. Chromate Fixatives
c. Lead Fixatives
CYTOLOGICAL FIXATIVE
1. Nuclear fixative – pH < 4.6 with gl.AA
(glacial acetic acid)
a. glacial acetic acid – can dissolve
content or fixative + stain
2. Cytoplasmic Fixative – pH > 4.6 without
gl.AA (glacial acetic acid)
 Osmic Acid
- Fixative for fats or lipids.
-2 in 1 because it is fixative and
at the same time stain.
- Gray and black = fat
-Amount is <10x
 Picric acid
- Fixative and at the same time
stain.
- For glycogen
- Yellow = there’s a presence of
glycogen
 B. Metallic Fixative OTHER OR COMMON PREPARATION
OF FIXATIVE UNDER ALDEHYDE
1. Mercurial
2. Lead 1. Formalin or formaldehyde
3. Chromate
4. Aldehyde 2. LILLIES- made of formalin + calcium
5. Alcohol acetate.
6. Acid
7. Metallic 3. 10% Formol saline – made of formalin +
sodium chloride.
- It preserved the morphology of the
ADVANTAGES OF FORMALIN whole brain or CNS and even the circle
of willies (hung brain or cord).
1. Cheap, stable and easy to prepare
2. Compatible with several stains 4. 10% buffered neutral formalin – made
3. Does not harden of formalin + disodium hydrogen
4. Best for CNS (Central Nervous System) phosphate (Na2HPO4) + sodium
5. Can preserve fats and lipids dihydrogen phosphate (NaH2PO4).

-best for fixing of iron, surgical in

DISADVANTAGES OF FORMALIN
1. Contact dermatitis
2. Asthma
3. Irritate the nose
4. It can cause lacrimation (increases of
tears)
5. Decrease ventilation to lungs
6. It has a risk factor to Lung CA (cancer)
7. It becomes formic acid upon oxidation
a. Dark brown pigment – Formalin
hematin pigment that can be
mistaken as malarial parasite or
melanin.
b. To prevent formic acid
oxidation – ADD 11 to 16% of
methanol
research specimen or post mortem
studies.
5. Kaisserlings compound – made of
formalin + potassium hydroxide (KOH) +
potassium nitrate (KNO3).
- Used for cadavers if the main purpose is
would last for several years.
6. Glutaraldehyde – special fixative and it
is made of 2 residues of formaldehyde.
- It is used for electron microscopy
examination.
- It is not suitable for immuno staining.

8. It would become paraformaldehyde or

polymerization – appearance is or turns
into forbid or milky
a. to prevent from polymerization
the remedy is FILTRATION.
METALS
1. Mercuric Chloride (HgCl2) – This would
react with acid or carboxyl.
Advantages of metal or mercuric
chloride

1. It is Metachromasia (introduce a
stain the color of the tissue is
different from the stain that is given
or added).

2. Good for trichrome staining.

3. It could fix connective tissue,

muscles, renal tissues and fibrin.
 4. Good for tissue photography. - Good for small pieces of liver, spleen,
connective tissue, fibers and nuclei.
5. Better nucleo and cytoplasmic
details. 2. Helly’s or known as Zenker-formol –
made of
6. It would precipitate all the protein.
H – HgCl2
7. It has the greater affinity to acid G – Glacial Acetic Acid
dyes. K – K2Cr4O7
N – NaSO4
F – Formalin
Disadvantages of metal or mercuric
chloride - Good for blood forming organs (Bone
marrow) or blood containing tissue.
1. It could cause hard and brittle - Bone marrow specimens
tissue.
2. It could cause lysis of RBC. 3. Schauddin’s – made up of HM.
3. There would be marked shrinkage.
4. Decrease the amount of -HM is made of HgCl2 + methanol
demonstrable glycogen. -good for smears of loose connective
5. Produces the fuse black granules. tissue or blood.
(What to do to remove the fuse
black granules: washing with lugol’s 4. Heidenhain’s Susa - made up of
iodine and 5% sodium thiosulfate.
6. It is unstable in the presence of - Is for skin biopsies and cytologic
glacial acetic acid and specimens.
formaldehyde.
H – HgCl2
G – Glacial Acetic Acid
REMOVAL OF FORMALIN K – K2Cr4O7
PIGMENTS N – NaCl

1. Kardasewitch’s method =
ammonia water + 70% ROH.
2. Lillies Method – Ammonia water +
70% ROH + H2O2 + acetone.
POTASSIUM DICHROMATE (K2Cr4O7) – This
would react both acid and basic reagents or
the tissue.

- Fastest method of removal of formalin ADVANTAGES OF POTASSIUM DICHROMATE

pigments . (K2Cr4O7)

1. Chromaffin tissue
3. Picric acid – saturated picric acid.
2. Colloid containing tissue (ex: testes
and thyroid)
3. Good for RBC
PREPARATIONS UNDER MERCURIC
4. Good for Mitochondria
CHLORIDE (HgCl2)
5. Good for Golgi apparatus
6. Good for Mitotic figures
1. Zenker’s – made of

H – HgCl2
G – Glacial Acetic Acid
K – K2Cr4O7
N – NaSO4

- Will give brown color.

 DISADVANTAGES OF POTASSIUM PREPARATIONS UNDER PICRIC ACID
DICHROMATE (K2Cr4O7)
1. Bouin’s – made of

1. Unstable - Best for embryos

2. It could cause a tissue to become hard
and brittle. F- formalin
3. There would be marked shrinkage of P- Picric Acid
tissue.
4. Blacken upon long standing. G – Glutaraldehyde
5. It would bleaches pigments.
6. Prolong used may lysis the RBC.
2. Brasil’s – Made of
PREPARATIONS UNDER POTASSIUM
- Best fixative for glycogen.
DICHROMATE (K2Cr4O7)
- Initiates decalcification.

F- formalin
1. Orth’s - made of 2.5 % K2Cr4O7
+ HCHO (formalin) + NaSO4. P- Picric Acid

G – Glutaraldehyde
2. Regaurd’s (Mollers) - made of 3%
A - alcohol
K2Cr4O7 + HCHO.
T- Trichloroacetic acid

ACIDS
3. Gendre’s - made of

F- formalin
1. Picric Acid – made of 2,4,6
trinitrophenol. P- Picric Acid

G – Glutaraldehyde
- Aqueous saturated solution.
A – alcohol

ADVANTAGES OF PICRIC ACID

OSMIC ACID (OSO4)
1. Small fragments of tissue.
2. Glycogen
3. It is very stable or the most stable acid.
ADVANTAGES OF OSMIC ACID

1. Electron microscopy
DISADVANTAGES OF PICRIC ACID 2. Chromosome
3. Myelin
4. Fats or lipids
1. Messy
2. Brittle and hard tissue
DISADVANTAGES OF OSMIC ACID
3. Marked shrinkage
4. Lysis RBC
1. Expensive
5. Yellow discoloration (remedy: wash
2. It could cause conjunctivitis which
with lithium carbonate and 5%
may lead to blindness.
sodium pyrosulfate.)
 GLACIAL ACETIC ACID – Hypotonic 3. Newcomer’s – made of
solution.
-best for nuclear protein.
-This cannot be use solely. (we cannot P – Propionic acid
use glacial acetic acid usually this is
inconjunction or incombination with I – Isopropyl Alcohol
other to neutralize the effect.)
G – Glacial Acetic Acid

ANOTHER FIXATIVE: ALCOHOL

4. Alcoholic formalin (Gendre’s Fluid)
1. This would act as fixative by –best for glycogen and sputum
disrupting the H one. specimen.
2. 80 -100% concentration to avoid
lysis of RBC.

ADVANTAGES OF ALCOHOL

1. Good for enzyme

2. Good for blood smears
(methanol)

DISADVANTAGES OF ALCOHOL

1. Distortion of morphology
2. It may cause a hard tissue
3. Cause shrunken in tissue

PREPARATIONS UNDER ALCOHOL

1. Carnoy’s – made of

- Faster in fixing
- Initiate dehydration

C – Chloroform

A – Alcohol

G- Glacial acetic acid

2. Clarke’s – made of

- Ideal for smears

- Sub Cultures

M- methanol

G- Glacial acetic acid