PART 2 - HP - PT Flashcards
most informative type of microscopy because it provides ultra structural details of glomerular basement membrane?
EM
H&E Staining End-Color Results
pale pink/red/ LR
Proteins in Edema
Cytoplasm
H&E Staining End-Color Results
pink?
Decalcified bone
Osteoid
Collagen
H&E Staining End-Color Results
deep pink
Muscle Fiber
H&E Staining End-Color Results
purplish pink?
Plasma cells
Osteoblast
Basophilic cytoplasm
Calcium
Calcified bone
H&E Staining End-Color Results
bright-orange to red?
RBC
Eosinophil granules
Keratin
H&E Staining End-Color Results
dark blue?
karyosome
H&E Staining End-Color Results
blue to blue-black
nucleus
H&E Staining End-Color Results
light blue to dark blue (ehrlicks hematoxylin)
cartilage
Decontamination of SCRAPIE, what to do?
Treatment: Immerse the tissue for
48 hrs in FORMALIN
1 hr in FORMIC ACID
48 hrs in FORMALIN
Accidental skin contact with hazardous chemicals, what to do?
wash with running tap water (15-30 mins)
never neutralize the sol’n
Eyes splashed with chemicals, what to do?
go to nearest eye wash station, use eye warm bottle for 15-30 mins
If an important slide is broken and replacement is not available, the section (if still intact) may be
transfer to another slide/ mount on another slide
it permits the entry of fluids into the cells/tses (makes tissues permeable to reagents)
a good fixative
Should be packed with cotton soaked in fixative or completely opened before being immersed in adequate fixing solution.
fixation of hollow organs: liver, stomach intestines
-most rapid (reduces 25-75% of impregnation time)
-impregnation under negative atmospheric pressure
-reco for urgent biopsies and delicate tissues
vacuum embedding
An element, which when added to a dye, forms a combination sometimes called a “lake” which forms a union between the stain and the tissue
mordant
Metallic mordants:?
“TAIL-CMC”
Tungsten
aluminum
iron
lead
chromium
molybdenum
copper
not a Metallic mordant:?
iodine
A mordant in combination with a dye forms a?
or
it combines with the tissue to form an insoluble tse - mordant dye complex)
lake
A dye solution that is applied after the primary stain has acted
counterstain
Cytoplasmic counterstains- RED?
“PEER”
phloxine B
eosin Y
eosin B
rose bengal
Cytoplasmic counterstains- YELLOW?
“OPO”
OG-6
Picric acid
Orange green
Cytoplasmic counterstains- GREEN?
lissamine green
light green SF
Nuclear counterstains- RED?
“CNS”
carmine
Neutral red
Safranin O
Nuclear counterstains- BLUE?
“MTCH”
methylene blue
toluidine blue
celestine blue
hematoxylin
most commonly used nuclear counterstain?
hematoxylin
most commonly used cytplasmic counterstain?
eosin Y
composition of Papanicolaou stain
“HOE”
hematoxylin
OG-6
Eosin azure
composition of Papanicolaou stain that stains nuclear structures?
hematoxylin
composition of Papanicolaou stain that stains cytoplasm of mature superficial cells?
OG-6
composition of Papanicolaou stain that stains cytoplasm of immature cells (Intermediate & parabasal cells)
Eosin azure
composition of eosin azure?
eosin
Bismarck brown
lithium carbonate
PTA
light green stain (36,50,65)
in Mod. Pap’s, it is omitted because it stains NOTHING
Bismarck brown
PAPANICOLAU METHOD OF REPORTING
what class?
-Absence of atypical/abnormal cells
-Normal Cytology
-NORMAL
class I
PAPANICOLAU METHOD OF REPORTING
what class?
-atypical cytology but no evidence of malignancy
-26-49% Atypical & normal cytology
-(Infection or inflammation)
-NORMAL & ATYPICAL
class II
PAPANICOLAU METHOD OF REPORTING
what class?
-suggestive of but NOT conclusive for malignancy
-Doubtful cytology
-50% undifferentiated cells (Regressive alteration of adult cell)
-SUSPICIOUS
class III
PAPANICOLAU METHOD OF REPORTING
what class?
-cytology strongly
-Suggestive of malignancy
-Frankly malignant cytology
-51-75% undifferentiated (Early stage of cancer)
-SUGGESTIVE
class IV
PAPANICOLAU METHOD OF REPORTING
what class?
-cytology conclusive for malignancy
-76-100% undifferentiated (With metastasis)
-INDICATIVE
class V
Glycogen stains
PAS stains what color?
magenta red
bright red
purple
Glycogen stains
The PAS reaction will demonstrate fungi, because the cell wall contains
chitin
Glycogen stains
Best Carmine stains what color?
bright red
Glycogen stains
Control material to be used with best carmine technique can usually found in the
liver
Glycogen stains
Langhan’s lodine stains what color?
mahogany brown
Oil soluble dyes
it has the greatest affinity to phospholipids among all the fat stains and gives triglycerides black color
Sudan black B
Oil soluble dyes
Recommended for neutral fats (triglycerides, lipids)
sudan IV
scharlach R
oil red o (ORO)
color: scarlet red
Oil soluble dyes
A good stain for the CNS
sudan III
example of BLUEING AGENTS?
1% alcoholic ammonia (ammonia water(
1 % aqueous lithium carbonate
Scott’s tap water
0.2-0.5% Bicarbonate
Potassium or Sodium acetate
Potassium or Sodium hydroxide
SPECIFIC STAINS
- it stains DNA?
- it stains RNA?
- methyl green (slightly acid pH)
& Feulgen (sugar) - pyronin
SPECIFIC STAINS
stains for muscle striations & demonstrate phosphat?
methyl green
SPECIFIC STAINS
stains MUCIN?
-PAS
-ALCIAN BLUE
-METACHROMATIC DYES (Toluidine blue & azure A)
-COLLOIDAL IRON
SPECIFIC STAINS
stains reticulin fibers (argyrophilic)?
Gomori’s silver impregnation
SPECIFIC STAINS
stains elastic fibers?
orcein (brown/DB)
SPECIFIC STAINS
most suitable for staining inflammatory cells, bacteria, blood parasites and some spirochetes
geimsa stain
SPECIFIC STAINS
A technique for demonstrating calcium, wherein sections are immersed in silver nitrate solution are exposed to bright light
von kossa
SPECIFIC STAINS
The pigment that is formed following a reaction of Ferrous iron from with potassium ferricyanide is known as
Turnbull’s blue
SPECIFIC STAINS
stains Ferric iron/Ferrous cyanide?
Perl’s Prussian blue
SPECIFIC STAINS
for acid mucopolysaccharides and is specific for connective tissue and epithelial mucin
alcian blue
SPECIFIC STAINS
Purpose of crystal thymol or a few drops of chloroform in alcian blue solution?
prevent mold formation/growth of microorgs
SPECIFIC STAINS
Staining Method for Basic Proteins?
alkaline fast green (color green) STAINS the nuleus (histones & protamines)
Chemical in the lab associated with bone marrow aplasia
Benzene (Clearing agent)
agents that are potentially explosives?
phenol
picric acid
silver salts
sodium azide
dioxane
Explosive when dry or mixed with metals. Toxic when absorbed through skin.
picric acid
Safe when fresh but explosive when old. Skin, eye and GIT irritant.
silver salts
Very toxic and fatal if swallowed or absorbed through the skin. Explodes upon contact with metals.
sodium azide
Corrosive to most metals. Should not be used in tissue processors. Skin and eye irritant and can cause severe GIT problems if ingested.
zinc chloride
Most commonly used FIXATIVE?
10% NEUTRAL BUFFERED FORMALIN
Most commonly used FIXATIVE FOR ELECTRON MICROSCOPE (EM) & HISTOCHEMISTRY
GLUTARALDEHYDE
Most commonly used FREEZING AGENT
LIQUID NITROGEN
Most commonly used DECALCIFYING AGENT
NITRIC ACID
Most commonly used DEHYDRATING AGENT
ETHYL ALCOHOL / ETHANOL
Most commonly used CLEARING AGENT
XYLENE
Most commonly used IMPREGNATING MEDIUM
PARAFFIN WAX
Most commonly used MICROTOME
ROTARY
Most commonly used ADHESIVE AGENT
MAYER’S EGG ALBUMIN
Most commonly used STAIN IN HISTOPATHOLOGY LABORATORY
HAND E STAIN
Most commonly used STAIN FOR CYTOLOGY
PAPANICOLAOU STAIN
Most commonly used MOUNTING MEDIUM
CANADA BALSAM
REMEDIES FOR BLACK DEPOSITS IN HISTOPATH
MERCURIC CHLORIDE black deposits
Remove with ALCOHOLIC IODINE
the process is called DEZENKERIZATION
REMEDIES FOR BLACK DEPOSITS IN HISTOPATH
OSMIUM TETROXIDE black deposits
Remove with TAP WATER
REMEDIES FOR BLACK DEPOSITS IN HISTOPATH
FORMALIN PIGMENT (Acid Formaldehyde Hematin)
Remove with PICRIC ACID (most common reagent)
it should not be neutralized as this may explode
Formalin
it is highly explosive when dry
Picric acid
it may explode upon air exposure
Dioxane
Metachromatic components?
“CECAMM”
cartilage
epithelial muscle
connective tissue
amyloid
mast cell
mucin
When examining an H&E-stained bone under the microscope, the technologist notices that there are many dark blue/purple (purplish blue) staining areas. What could this mean?
Bone was underdecalcified
In H and E staining, insufficient time in the acid differentiator will cause:
Under-differentiation
Epithelial filament markers:
Keratin
Epithelial membrane antigen
Carcinoembryonic membrane antigen
Thyroid transcription factor-1
Prostate specific antigen
It is a component of B5 fixative
Mercuric Chloride
Antigens are most likely to be demonstrated at max sensitivity with what?
Frozen sections fixed in acetone
What would be proper fixation procedure for a large tissue?
Large tissue must be cut into smaller pieces
What is the most accurate way to determine the end point of decalcification?
Radiography
What is the purpose of placing the tissue ribbon in the flotation water bath?
To flatten the tissue section
Why should gloves be worn when cutting tissues on the microtome?
To prevent skin cells from getting on the microscope slide
The main difference between progressive and regression staining is the presence of?
differentiator
characteristics of osmium tetroxide?
Used mostly for EM
Fixes lipids
Commonly a secondary fixative
Skin tumor biopsies are preserved with what fixative?
Heidenhain’s Susa
it is an ideal embedding medium for undecalcified bone and other hard tissue
methyl methacrylate
it is a popular embedding medium for light microscopy that is highly hydrophilic.
polyglycol methacrylate
Airholes found in the tissue during trimming is due to?
Incomplete infiltration
Disadvantages of celloidin impregnation?
Slow and tedious
Serial sections are difficult to prepare
Very thin sections are difficult to cut
When cutting through hard tissues such as thyroid and cervix, the ideal cutting stroke is:
Firm, quick stroke
to evenly cut tse
A dehydrating agent that is known to be an eye and skin irritant and may cause conjunctival irritation:
Tetrahydrofuran
