HTMLE DIAGNOSTIC

EXAM RATIO
--KLUBSYBEAR--
 RITM National reference laboratory for dengue, influenza, TB, and other
#3 mycobacteria, malaria and other parasites, bacterial enteric diseaeses,
measles and other viral exanthems, mycology, enterovirus, antimicrobial
resistance and emerging diseases.
NRL for confirmatory testing of blood units

PHIL. LUNG National reference laboratory for biochemistry

CENTER
NKTI National reference laboratory for hematology including immunohematology,
immunopathology and anatomic pathology

SLH National reference lab for HIV/AIDS, hepatitis, syphilis, and other STD
#8

FIXATIVE 10-20 Times the volume of the tissue

except osmium tetroxide (5-10x) .
DECALCIFYING AGENT ≥20 times the volume of the tissue
DEHYDRATING AGENT ≥10 times the volume of the tissue
CLEARING AGENT ≥10 times the volume of the tissue
IMPREGNATING AGENT ≥25 times the volume of the tissue
 #9
TISSUE SECTION THICKNESS
Paraffin (routine) 4-6 u (micra)
Celloidin section 10-15 u (micra)
Frozen section 10 u (micra)
Ultrathin section Semithin = 0.5 to 1u (micra)
Ultrathin = 50-120nm or 500-1200 angstrom
Electron microscopy section 0.5u (micra)
#10

1.Picric Acid Saturated Alcoholic solution of picric acid

2.Kardesawitsch’s method Ethyl alcohol, ammonia water
3.Lillie’s method Acetone, hydrogen peroxide, ammonia water
 #11

Fixation is retarded by Large tissue, presence of

mucus, fat , blood, cold
temp
Fixation is enhanced by Smaller and thinner tissues,
agitation and heat(37-56oC)
#12

Bevel Angle Angle formed between the cutting edges

of the knife.
27-32 angle
Clearance Angle Angle formed between the cutting edge
of the knife and the tissue block.
5-15 angle
Optimum cutting angle 15o
 #13

HONING Sharpening Removal of Heel to toe

gross nicks direction
STROPPING Polishing Removal of Toe to heel
burrs direction
#15

Biconcave knife 120 mm in length Recommended for cutting paraffin embedded sections
on a rotary microtome
Plane concave knife 25 mm in length Less concave sides are recommended for cutting
celloidin-embedded tissue blocks on a sliding
microtome. More concave sides are used to cut
paraffin sections on base-sledge, rotary or rocking
microtome
Plane wedge knife 100 mm in length Recommend for frozen sections or for cutting
extremely hard and tough specimens embedded in
paraffin blocks, using a base sledge type or sliding
microtome
#18 PYKNOSIS Reduction and
condensation of nucleus
Characterized by nuclear shrinkage and
increased basophilia. The chromatin condenses
into a solid, shrunken basophilic mass.

KARYORRHEXIS Fragmentation of nucleus The pyknotic nucleus undergoes fragmentation.

With the passage of time (a day or two), the
nucleus in the necrotic cell totally disappears.

KARYOLYSIS Dissolution of nucleus The basophilia of the chromatin may fade

(karyolysis), a change that presumably reflects
loss of DNA because of enzymatic degradation
by endonucleases.
 TYPE DESCRIPTION
COAGULATIVE NECROSIS A form of necrosis in which the architecture of deadtissues is preserved for a

#22 must know!!

span of at least some days. Ischemia caused byobstruction in a vessel may lead
to a coagulative necrosis of the supplied tissue inall organs except the brain.

NOTE: Infarct – a localized area of coagulative necrosis

LIQUEFACTIVE NECROSIS Characterized by digestion of dead cells, resulting intransformation of the

tissue into a liquid viscous mass.

NOTE: The necrotic material is frequently creamy yellow because of the

presence of dead leukocytes and is called pus.

GANGRENOUS NECROSIS Not a specific pattern of cell death; usually appliedtoo limb, generally the
lower leg, that has lost its blood supply and has undergonecoagulative
necrosis.
 Refers to the cheese-like friable white appearance of thearea of
CASEOUS NECROSIS necrosis; Often seen in tuberculosis

NOTE: Granuloma – Necrotic area appears as structureless

collection offragmented or lysed cells and amorphous granular debris
enclosed within adistinctive inflammatory border on microscopic
examination.
It refers to focal areas of fat destruction, typically resulting
FAT NECROSIS fromrelease of activated pancreatic lipases into the substance of the
pancreas andperitoneal cavity.

NOTE: On histologic examination, the necrosis takes the form of

foci of shadowy outlines of necrotic fat cells, with basophilic calcium
deposits, surrounded by an inflammatory reaction.
– A special form of necrosis usually seen in immunereactions
FIBRINOID NECROSIS involving blood vessels. This pattern of necrosis typically occurs
whencomplexes of antigens and antibodies are deposited in the walls
of the arteries.Deposits of these “immune complexes,” together with
fibrin that has leaked ofvessels, result in a bright pink and amorphous
appearance in H&E stains, called“fibrinoid” (fibrin-like) by
pathologists.
 #31 mga must know

Most commonly used antibody for IgG

Immunocytochemistry

The structural part of the antigen that reacts with Epitope (antigenic derminant)
an antibody.

The structural part of an antibody that reacts with paratope

an antigen.

Recommended thickness of tissue sections for 3-4u (micra)

Immunohistochemistry
 #36
SKELETAL MUSCLE CARDIAC MUSCLE SMOOTH MUSCLE
Very long, cylindrical, Elongated, BRANCHED, Fusiform cells, SPINDLE-
MULTINUCLEATED UNINUCLEATED (central) SHAPED, UNINUCLEATED
(peripheral), that show CROSS- cells, with CROSS- (central), DO NOT SHOW
STRIATIONS STRIATIONS CROSS-STRIATIONS in the
light microscope (No Striations)

Has INTERCALATED DISK,

a special structure found only in
Cardiac muscles
VOLUNTARY CONTROL INVOLUNTARY CONTROL INVOLUNTARY CONTROL
 #37
VIRCHOW Organs are removed one by one

ROKITANSKY “In-situ” dissection; in part combined with the removal of organ

blocks
Dissection while organs are in place.
GHON “En Bloc” removal;
LETULLE “En Masse” removal; recommended for pediatric patient
#40 MUST KNOW
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