Trim - Section - Stain Flashcards
Tissues must be trimmed by at least __ mm of wax
2 mm
Trimming using a knife or a blade forming truncated pyramind/ 4 sided prism
Coarse trimming
Trimming where block is placed in the microtome setting it at 15 mm
Fine trimming
Sectioning thickness for paraffin
4-6 um
Sectioning thickness for celloidin
10-15 um
Sectioning thickness for frozen
10 um
Sectioning thickness for semi thin (glass)
0.5-1.0 um
Sectioning thickness for ultrathin (diamond)
500-1200 Å or 50-120 um
Section cut is picked up by:
Index finger
Camel hairbrush
Spatula
Flat beaded forceps
What to do to tissues that tend to crumble
Exhaling gently (reduces static electricity)
A __ & __ cutting stroke produces the best results & least compression
slow & uniform
Sections are removed in ribbons of __
10
Fishing out is done with a clean slide in a __
vertical position
most commonly used adhesive & its composition
Mayer’s egg albumin
- egg whites
- glycerin (⬆️ viscosity & no drying)
- thymol crystals (preservative)
Adhesive for cytology
APES
3-aminopropyltriethoxysilane)
Adhesive for IHC
Poly-L-Lysine
Done to study the architectural pattern of tissue & to enhance contrast
Staining
Methylene blue is an example of a __ stain
Direct
Gram stain is an example of a __ stain
Indirect
Serves as a link between tissue & dye; an integral part of staining rxn
Mordant
Hastens the speed of staining rxn but not essential
Accentuator
Overstaining & the excess is removed
Regressive staining (H&E stain)
Staining in definite sequence, once its stained its not washed/decolorized
Progressive stain
Primary dye is a basic dye & the counterstain is acidic (vice versa)
Differential staining
Tissues are stained in color shades that are SIMILAR to the color of the dye itself
Orthochromic
Staining with a color that is DIFFERENT from that of the stain itself
Metachromatic staining
Reduced by argentaffin cells forming black deposits under microscope
Ammoniacal silver
Blueing agents
- tap h2o
- sat. lithium carbonate
- 0.5% ammonia in dist. h2o
- ammonium hydroxide
- scott’s soln
Examples of vital staining done by injecting the dye
lithium
india ink
carmine
Example of vital stains done after removal from the living body
neutral red
janus green
trypan blue
thionine
nile blue
toluidine blue
Red cytoplasmic counterstain
Eosin Y
Eosin B
Phloxine B
Yellow cytoplasmic counterstain
Picric acid
Orange G
Rose bengal
Green cytoplasmic counterstain
Light green
Lissamine green
Red NUCLEAR counterstain
neutral red
safranin O
carmine
hematoxylin
Blue NUCLEAR counterstain
methylene blue
toluidine blue
celestine blue
Natural biological stains
Cochineal
logwood
veggie extracts
Synthetic biological stains
Aniline
Coal tar
Oxidation of hematoxylin
Ripening
Ripening that exposes the substance to air/sun for 4 months
Natural ripening
Accelerates ripening process
Artificial ripening
Hematoxylin extracted from the core wood of a mexican tree called
Haematoxylon campechianum
Active coloring agent of hematoxylin
Hematein
oxidation leading to production of other useless compounds
Over-ripening