HISTO LAB MIDTERM

Question 1

WHAT IS THE ROLE OF HARRIS HEMATOXYLIN IN H&E

Answer 1

Stains nuclei

Question 2

WHAT IS THE ROLE OF 0.5% ACID ALCOHOL IN H&E, HOW DOES IT WORK?

Answer 2

Differentiator works by breaking links between the tissue & mordant

Question 3

WHAT IS THE ROLE OF EOSIN?

Answer 3

ACTS AS A COUNTERSTAIN. STAINS ALL TISSUE COMPONENTS EXCEPT FOR NUCLEI

Question 4

WHAT IS THE OXIDIZING AGENT IN H&E

Answer 4

NalO3 –Sodium iodate

Question 5

WHAT IS THE PRINCIPLE OF H&E?

Answer 5

IONIC BONDING

Question 6

IS H&E REGRESSIVE OR PROGRESSIVE?

Answer 6

REGRESSIVE

Question 7

WHAT COLOURS DOES H&E STAIN?

Answer 7

STAINS NUCLEI PURPLE, STAINS MUSCLE, RBCS, CYTOPLASM PINK

Question 8

H&E: NUCLEI PALE, WHY?

Answer 8

CAN BE FROM HEMATOXYLIN BEING OLD & OVER OXIDIZED. CAN ALSO BE FROM OVER-DIFFERENTIATION IN 0.5% ACID ALCOHOL.

Question 9

MASSONS TRICHROME HEMATOXYLIN USED & ITS ROLE

Answer 9

WEIGERT’S, STAINS NUCLEI BLACK

Question 10

IS MT PROGRESSIVE OR REGRESSIVE?

Answer 10

PROGRESSIVE

Question 11

DOES MT HAVE A STABILIZING AGENT?

Answer 11

NO

Question 12

OXIDIZING AGENT IN MT

Answer 12

FECL3

Question 13

ROLE OF BEBIRCH SCARLET AND ACID FUSION, WHAT TECHNIQUE ARE THEY USED IN?

Answer 13

BOTH ACT AS A COUNTERSTAIN
SCARLET– SMALL, STAINS BASIC TISSUE COMPONENTS
AF–LARGE AND STAINS COLLAGEN RED

Question 14

PTA/PMA ROLE IN MT

Answer 14

ACTS AS A DIFFERENTIATOR, REMOVES RED COLOUR OF AF FROM COLLAGEN

Question 15

ANILINE BLUE, ROLE & ITS PROBLEMS

Answer 15

COUNTERSTAIN FOR MT, STAINS COLLAGEN BLUE, TOO LONG IN THIS DYE CAN RESULT IN A MUDDY CYTOPLASM

Question 16

1% ACETIC ACID ROLE & PROBLEMS

Answer 16

REMOVES EXCESS ANILINE BLUE FROM THE BACKGROUND. TOO LONG IN AA RESULTS IN PALE COLLAGEN, TOO SHORT IN AA RESULTS IN MUDDY CYTOPLASM

Question 17

WHAT COLOUR DOES MT STAIN?

Answer 17

STAINS NUCLEI DARK GREY, STAINS MUSCLES, CYTOPLASM AND RBCS RED, COLLAGEN STAINS BLUE

Question 18

WHAT IS THE ROLE OF BOUINS IN MT

Answer 18

PRE-MORDANT THE TISSUE TO INCREASE STAINING OF COLLAGEN IF FIXED IN 10% NBF. STAINS TISSUE COMPONENTS YELLOW.

Question 19

WHAT IS THE PRINCIPLE OF MASSONS TRICHROME

Answer 19

POROSITY

Question 20

GOMORI’S ALDEHYDE FUSCHIN TECHNIQUE PRINCIPLE

Answer 20

HYDROGEN BONDING

Question 21

WHAT COLOUR DOES GAF STAIN?

Answer 21

ELASTIC= PURPLE, COLLAGEN=RED, OTHER=YELLOW

Question 22

ROLE OF ALDEHYDE FUCHSIN

Answer 22

STAINS ELASTIC VIA HYDROGEN BONDING, STAINS EVERYTHING PURPLE

Question 23

70% ETHANOL ROLE & PROBLEMS

Answer 23

ACTS AS A WASHING AGENT IN GAF, REMOVES EXCESS AF FROM THE BACKGROUND. TOO SHORT OF A WASH CAN RESULT IN A MUDDY CYTOPLASM.

Question 24

VAN GIESON REAGENT, ROLE & WHAT TECHNIQUES ITS USED IN

Answer 24

ACTS AS A COUNTERSTAIN, ACID FUCHSIN= ACID DYE, LARGE, STAINS COLLAGEN RED. PICRIC ACID=ACID DYE, SMALL, STAINS YELLOW
USED IN GAF & VERHOFFS

Question 25

VERHOFF'S VAN GIESON TECHNIQUE PRINCIPLE

Answer 25

HYDROGEN BONDING

Question 26

WHAT HEMATOXYLIN DOES VVG USE & ROLE?

Answer 26

VERHOFF'S HEMATOXYLIN (IRON), STAINS EVERYTHING BLACK

Question 27

ROLE OF FECL3 IN VVG

Answer 27

ACTS AS A MORDANT, OXIDIZING AGENT AND DIFFERENTIATOR

Question 28

ROLE OF IODINE

Answer 28

ACCOMPLISHES THE FIRST STEP OF DEZENKERIZATION--THE REMOVAL OF MERCURIC PIGMENT

Question 29

SODIUM THIOSULPHATE ROLE IN VVG

Answer 29

BACKGROUND CLEARING, ACCOMPLISHES THE SECOND STEP IN DEZENKIRIZATION. MAKES THE BACKGROUND PALE GREY, MUST BE WASHED WELL

Question 30

WHAT COLOUR DOES VVG STAIN THE COMPONENTS?

Answer 30

ELASTIC-BLACK, NUCLEI=BLACK, RBC=BRIGHT YELLOW, COLLAGEN=RED, EVERYTHING ELSE=YELLOW

Question 31

G&S PRINCIPLE

Answer 31

STAINS VIA ARGYROPHILLIC SILVER IMPREGNATION

Question 32

WHAT IS OXIDIZING AGENT IN G&S & ROLE?

Answer 32

POTASSIUM PERMAGANATE, STAINS TISSUE COMPONENTS BROWN

Question 33

WHAT IS ROLE OF OXALIC ACID

Answer 33

ACTS AS A BLEACHING AGENT, REMOVES DISCOLOURATION OF POTASSIUM PERMANGANATE (TISSUES MUST BE PALE TO PROCEED)

Question 34

FERRIC AMMONIUM SULPHATE/IRON ALUM ROLE

Answer 34

SENSITIZING AGENT, CAUSES RETICULIN FIBRES TO BE COATED WITH FE3 IONS

Question 35

AMMONIACAL SILVER SOLUTION ROLE

Answer 35

SILVER IMPREGNATION OF RETIC RIBERS, FE3 ARE REPLACED BY SILVER DIAMINE IONS

Question 36

10% FORMALIN ROLE

Answer 36

ACTS AS A REDUCING AGENT, CONVERTS AMMONIACAL SILVER IONS IN METALLIC SILVER, RETICULIN FIBRES = DARK BROWN

Question 37

WHAT IS THE CAUSE OF SILVER PRECIPITATE IN BACKGROUND?

Answer 37

Improper washing, sodium thiosulphate omitted, time in gold chloride too short, use of metal forceps or dirty glassware

Question 38

GOLD CHLORIDE ROLE

Answer 38

TONING AGENT, GOLD IONS REPLACE SILVER IONS TO PROVIDE GREATER CONTRAST UNDER THE MICROSCOPE

Question 39

SODIUM THIOSULPHATE ROLE IN G&S

Answer 39

ACTS AS A FIXING AGENT (STOPS ALL CHEMICAL ACTIVITY), REMOVES ANY EXCESS AG AND AU

Question 40

NEUTRAL RED

Answer 40

COUNTERSTAIN, STAINS NUCLEI AND BACKGROUND

Question 41

WHAT COLOURS DOES G&S STAIN?

Answer 41

RETIC FIBRES= BLACK, NUCLEI= RED, OTHER= PINK

Question 42

ROLE OF PERIODIC ACID

Answer 42

PRIMARY OXIDIZING AGENT (OXIDIZES ONLY TO THE ALDEHYDE STAGE)

Question 43

ROLE OF SCHIFF'S REAGENT

Answer 43

BINDS TO REACTIVE DIALDEHYDE SITES IN TISSUE

Question 44

RUNNING TAP WATER ROLE IN PAS

Answer 44

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

Question 45

HEMATOXYLIN ROLE IN G&S

Answer 45

HARRIS OR MAYERS USED, STAINS NUCLEI BLUE

Question 46

COLOURS OF PAS SLIDE

Answer 46

BRIGHT PINK= NEUTRAL MUCINS, GLYCOGEN, COLLAGEN, BASEMENT MEMBRANE, CARTILAGE, AMYLOID DARK BLUE= NUCLEI OTHER= LIGHT PINK/PURPLE

Question 47

PRINCIPLE OF PAS

Answer 47

HISTOCHEMICAL REACTION

Question 48

ALCIAN BLUE ROLE

Answer 48

STAINS ACID MUCINS (BLUE)

Question 49

ALCIAN BLUE 2.5 PH

Answer 49

SULPHATED & CARBOXYLATED

Question 50

ALCIAN BLUE 1.5 PH

Answer 50

SULPHATED ONLY

Question 51

ALCIAN BLUE STEPS IN DYEING

Answer 51

1. ALCIAN BLUE, 2 HARRIS HEMATOXYLIN (COUNTERSTAIN), 3. 0.5% ACID ALCOHOL (DIFFERENTIATOR), 4. AMMONIA WATER (BLUING AGENT)

Question 52

PASD SLIDE

Answer 52

AMYLASE ENZYME, REMOVES GLYCOGEN RESULTING IN A SLIDE WITH BLUE NUCLEI AND LIGHT BLUE/PURPLE BACKGROUND.