HISTO LAB MIDTERM Flashcards
WHAT IS THE ROLE OF HARRIS HEMATOXYLIN IN H&E
Stains nuclei
WHAT IS THE ROLE OF 0.5% ACID ALCOHOL IN H&E, HOW DOES IT WORK?
Differentiator works by breaking links between the tissue & mordant
WHAT IS THE ROLE OF EOSIN?
ACTS AS A COUNTERSTAIN. STAINS ALL TISSUE COMPONENTS EXCEPT FOR NUCLEI
WHAT IS THE OXIDIZING AGENT IN H&E
NalO3 –Sodium iodate
WHAT IS THE PRINCIPLE OF H&E?
IONIC BONDING
IS H&E REGRESSIVE OR PROGRESSIVE?
REGRESSIVE
WHAT COLOURS DOES H&E STAIN?
STAINS NUCLEI PURPLE, STAINS MUSCLE, RBCS, CYTOPLASM PINK
H&E: NUCLEI PALE, WHY?
CAN BE FROM HEMATOXYLIN BEING OLD & OVER OXIDIZED. CAN ALSO BE FROM OVER-DIFFERENTIATION IN 0.5% ACID ALCOHOL.
MASSONS TRICHROME HEMATOXYLIN USED & ITS ROLE
WEIGERT’S, STAINS NUCLEI BLACK
IS MT PROGRESSIVE OR REGRESSIVE?
PROGRESSIVE
DOES MT HAVE A STABILIZING AGENT?
NO
OXIDIZING AGENT IN MT
FECL3
ROLE OF BEBIRCH SCARLET AND ACID FUSION, WHAT TECHNIQUE ARE THEY USED IN?
BOTH ACT AS A COUNTERSTAIN
SCARLET– SMALL, STAINS BASIC TISSUE COMPONENTS
AF–LARGE AND STAINS COLLAGEN RED
PTA/PMA ROLE IN MT
ACTS AS A DIFFERENTIATOR, REMOVES RED COLOUR OF AF FROM COLLAGEN
ANILINE BLUE, ROLE & ITS PROBLEMS
COUNTERSTAIN FOR MT, STAINS COLLAGEN BLUE, TOO LONG IN THIS DYE CAN RESULT IN A MUDDY CYTOPLASM
1% ACETIC ACID ROLE & PROBLEMS
REMOVES EXCESS ANILINE BLUE FROM THE BACKGROUND. TOO LONG IN AA RESULTS IN PALE COLLAGEN, TOO SHORT IN AA RESULTS IN MUDDY CYTOPLASM
WHAT COLOUR DOES MT STAIN?
STAINS NUCLEI DARK GREY, STAINS MUSCLES, CYTOPLASM AND RBCS RED, COLLAGEN STAINS BLUE
WHAT IS THE ROLE OF BOUINS IN MT
PRE-MORDANT THE TISSUE TO INCREASE STAINING OF COLLAGEN IF FIXED IN 10% NBF. STAINS TISSUE COMPONENTS YELLOW.
WHAT IS THE PRINCIPLE OF MASSONS TRICHROME
POROSITY
GOMORI’S ALDEHYDE FUSCHIN TECHNIQUE PRINCIPLE
HYDROGEN BONDING
WHAT COLOUR DOES GAF STAIN?
ELASTIC= PURPLE, COLLAGEN=RED, OTHER=YELLOW
ROLE OF ALDEHYDE FUCHSIN
STAINS ELASTIC VIA HYDROGEN BONDING, STAINS EVERYTHING PURPLE
70% ETHANOL ROLE & PROBLEMS
ACTS AS A WASHING AGENT IN GAF, REMOVES EXCESS AF FROM THE BACKGROUND. TOO SHORT OF A WASH CAN RESULT IN A MUDDY CYTOPLASM.
VAN GIESON REAGENT, ROLE & WHAT TECHNIQUES ITS USED IN
ACTS AS A COUNTERSTAIN, ACID FUCHSIN= ACID DYE, LARGE, STAINS COLLAGEN RED. PICRIC ACID=ACID DYE, SMALL, STAINS YELLOW
USED IN GAF & VERHOFFS
VERHOFF’S VAN GIESON TECHNIQUE PRINCIPLE
HYDROGEN BONDING
WHAT HEMATOXYLIN DOES VVG USE & ROLE?
VERHOFF’S HEMATOXYLIN (IRON), STAINS EVERYTHING BLACK
ROLE OF FECL3 IN VVG
ACTS AS A MORDANT, OXIDIZING AGENT AND DIFFERENTIATOR
ROLE OF IODINE
ACCOMPLISHES THE FIRST STEP OF DEZENKERIZATION–THE REMOVAL OF MERCURIC PIGMENT
SODIUM THIOSULPHATE ROLE IN VVG
BACKGROUND CLEARING, ACCOMPLISHES THE SECOND STEP IN DEZENKIRIZATION. MAKES THE BACKGROUND PALE GREY, MUST BE WASHED WELL
WHAT COLOUR DOES VVG STAIN THE COMPONENTS?
ELASTIC-BLACK, NUCLEI=BLACK, RBC=BRIGHT YELLOW, COLLAGEN=RED, EVERYTHING ELSE=YELLOW
G&S PRINCIPLE
STAINS VIA ARGYROPHILLIC SILVER IMPREGNATION
WHAT IS OXIDIZING AGENT IN G&S & ROLE?
POTASSIUM PERMAGANATE, STAINS TISSUE COMPONENTS BROWN
WHAT IS ROLE OF OXALIC ACID
ACTS AS A BLEACHING AGENT, REMOVES DISCOLOURATION OF POTASSIUM PERMANGANATE (TISSUES MUST BE PALE TO PROCEED)
FERRIC AMMONIUM SULPHATE/IRON ALUM ROLE
SENSITIZING AGENT, CAUSES RETICULIN FIBRES TO BE COATED WITH FE3 IONS
AMMONIACAL SILVER SOLUTION ROLE
SILVER IMPREGNATION OF RETIC RIBERS, FE3 ARE REPLACED BY SILVER DIAMINE IONS
10% FORMALIN ROLE
ACTS AS A REDUCING AGENT, CONVERTS AMMONIACAL SILVER IONS IN METALLIC SILVER, RETICULIN FIBRES = DARK BROWN
WHAT IS THE CAUSE OF SILVER PRECIPITATE IN BACKGROUND?
Improper washing, sodium thiosulphate omitted, time in gold chloride too short, use of metal forceps or dirty glassware
GOLD CHLORIDE ROLE
TONING AGENT, GOLD IONS REPLACE SILVER IONS TO PROVIDE GREATER CONTRAST UNDER THE MICROSCOPE
SODIUM THIOSULPHATE ROLE IN G&S
ACTS AS A FIXING AGENT (STOPS ALL CHEMICAL ACTIVITY), REMOVES ANY EXCESS AG AND AU
NEUTRAL RED
COUNTERSTAIN, STAINS NUCLEI AND BACKGROUND
WHAT COLOURS DOES G&S STAIN?
RETIC FIBRES= BLACK, NUCLEI= RED, OTHER= PINK
ROLE OF PERIODIC ACID
PRIMARY OXIDIZING AGENT (OXIDIZES ONLY TO THE ALDEHYDE STAGE)
ROLE OF SCHIFF’S REAGENT
BINDS TO REACTIVE DIALDEHYDE SITES IN TISSUE
RUNNING TAP WATER ROLE IN PAS
ACTS AS A SECONDARY OXIDIZING AGENT–PRODUCES THE FINAL REACTION PRODUCT BY REMOVING THE SULFONIC ACID ION TO RESTORE BASIC CHROMOPHORE OF BASIC FUCHSIN (QUINOID STRUCTURE), WHEN TISSUE TURNS PINK
HEMATOXYLIN ROLE IN G&S
HARRIS OR MAYERS USED, STAINS NUCLEI BLUE
COLOURS OF PAS SLIDE
BRIGHT PINK= NEUTRAL MUCINS, GLYCOGEN, COLLAGEN, BASEMENT MEMBRANE, CARTILAGE, AMYLOID
DARK BLUE= NUCLEI
OTHER= LIGHT PINK/PURPLE
PRINCIPLE OF PAS
HISTOCHEMICAL REACTION
ALCIAN BLUE ROLE
STAINS ACID MUCINS (BLUE)
ALCIAN BLUE 2.5 PH
SULPHATED & CARBOXYLATED
ALCIAN BLUE 1.5 PH
SULPHATED ONLY
ALCIAN BLUE STEPS IN DYEING
- ALCIAN BLUE, 2 HARRIS HEMATOXYLIN (COUNTERSTAIN), 3. 0.5% ACID ALCOHOL (DIFFERENTIATOR), 4. AMMONIA WATER (BLUING AGENT)
PASD SLIDE
AMYLASE ENZYME, REMOVES GLYCOGEN RESULTING IN A SLIDE WITH BLUE NUCLEI AND LIGHT BLUE/PURPLE BACKGROUND.