KNIVES + ADHESIVES + RI Flashcards
LIGHT MCIROSCOPY
- for celloidin and paraffin embedding
- length
- microtome used - for frozen sections or extremely hard and tough specimens in paraffin
- length
- microtome used - paraffin section
- length
- microtome used
- knife used
- requires smaller _______ - wedge knife w/ steep cutting edge
- ________ knife
- Plane-Concave
- 25 mm
- sliding (base-sledge); rotary; rocking - Plane-Wedge
- 100mm
- sliding (base-sledge) - Biconcave
- 120mm
- rotary
- Heiffor knife
- clearance angle - Plane-Shaped
- tungsten carbide coated knife
ELECTRON MICROSCOPY
- resin blocks
- trimming or semi thin sectioning
OTHERS
1. sharp cutting edge producing 2-4um
- two blades
2. partially calcified tissues, paraffin and frozen section
- producing ___ sections
3. for cryostat
- producing ___ sections
ELECTRON MICROSCOPY
- Diamond-Edge
- Broken Glass Knives
OTHERS
1. Disposable Blades
- Platinum 6 & Chromium 7
2. Safety Razor Blades
- 10um & above
3. Teflon-coated Blades
- 2-4um
ANGLES DURING SECTIONING
- between the edge of the knife & the tissue block
- degrees - angle of cutting
- degrees - angle of cutting facet
- degrees
- cutting edge: more acute = _____
- ______ bevel angle = sharper - angle that’s outer edge of know w/ line that is 90
degrees of tissue block
- ___ rake angle & ___ clearance angle gives less compression to the tissue block
- Clearance Angle
- 5-10 degrees - Wedge Angle
- 15 degrees - Bevel Angle
- 27-32 degrees
- sharper
- smaller bevel angle - Rake Angle
- high rake angle & low clearance angle
SHARPENING OF MICROTOME KNIVES
1. doesn’t maintain the edge
2. is likely to nick against hard objects
3. tests in order to assess the sharpness of cutting edge
- should cut paraffin wax block at ______ w/o _____
- criterion w/ strong nearly lines, cutting edge will
show slight reflection as very narrow continuous straight line
- will split a hair down across it with only their _____
SHARPENING OF MICROTOME KNIVES
1. TOO SOFT
2. TOO HARD
3.
- 2-4um; SERRATIONS
- Von Mhol’s criterion
- resistance
HONING vs STROPPING
- removal of gross nicks & blemishes/ irregularities
- material
- best result; manual sharpening
- more polishing effect; to finish after coarse honing
- coarsest/ badly nickeled knives
- for coarse grinding & finishing - removal of burrs, for final polishing and sharpening of cutting edge
- material
- HONING
- smooth stones/oilstones
(8x3) & machine hone
- belgium yellow
- arkansas
- fine carborundum
- belgian jack vein (blue
green) - STROPPING
- shell horse leather
OTHER HONING PROCEDURES:
- excellent substitute for stone
- inches
- ____: used for final polishing
- similar to _______ - after repeated sharpening
- widened _____ (>__ degrees) - large circular glass plate - specific material
- Plate Glass Honing
- 8x3x1 in
- Diamantine
- Belgium yellow - Factory Grinding
- bevel angle (>35 degrees) - Automatic Hones
- Fanz Shandon-Elliot, autosharp III
GENERAL STEPS IN FIXING SECTIONS ONTO
SLIDE
_____: to straighten the tissue sections
- ____ or_____ degrees lower than MP of ____
- not longer than ____
- alternative is stretching sections in a slide with ____ or _____
____: transfer of tissue sections/ ribbons on to the slide
1. water bath should have temp ____ lower than MP of wax
_____: correct positioning of the tissue section/ribbon on the slide
Floating on waterbath
- 45-50Cor 6-10C; wax
- 30s
- 20% alcohol/ water
Fishing out
1. 10C
Orientation
GENERAL STEPS IN FIXING SECTIONS ONTO
SLIDE
_____: prevent washing out of tissue sections during staining
- substance which can be smeared onto the slides
so that sections stick well to the slides
-no need for staining solution
- need adhesive (5)
TYPES
2. most commonly used; easy, convenient & relatively inexpensive
- equal amount of
- pinch of ___ to avoid molds in albumin
3. sections are dried and stored in 70% alcohol until it is ready
- components
4. added to the waterbath during flotation rather than applying it on slides
- components
5. slides are coated and allowed to dry at 37C for one hour or overnight prior to use
6. widely used in immunohistochemistry
- aq detergent diluted to ______
7. very useful in cytology, preparations of proteinaceous/bloody material
- meaning
- percent
- particularly in
Adhesion
- Adhesives
- routine staining
- cryostat sections; immunocytochemistry; CNS tissues; tissues w/ blood clots; decalcified tissues;
TYPES
2. Mayer’s Egg Albumin
- egg white + glycerin
- thymol
3. Dried Albumin
- dried albumin + Sodium Chloride + thymol
4. 1% Gelatin
- gelatin + glycerol + phenol
5. Gelatin-Formaldehyde Mixture
6. Poly-L-lysine
- 0.01% concentration
7. APES
- 3-aminoprophylthriethoxysilane
- 2%
- cytospin
GENERAL STEPS IN FIXING SECTIONS ONTO
SLIDE
Deparaffination & Drying Sections: temp for time
- wax oven
- incubators
- hot plate
- delicate tissues
- blower-type electric slide dryer
- 56C– 60C for 2 hrs
- overnight
- 45C – 55C for 30 – 45 mins
- 37C for at least 24 hours
- 50 – 55C for 20–30 mins
GENERAL STEPS IN FIXING SECTIONS ONTO
SLIDE
___ secondary fixation
- ______ for 24hrs
- used primarily as ____ & ______
- 5-10 mins in either: (2)
Post-mordanting
- 2.5 – 3% Potassium Dichromate
- mordant & secondary as fixative
- aq. sol’n. of mercury chloride
- aq. picric acid
CELLOIDIN SECTIONS
1. ___ um
2. ____ is not required
3. ____ method: store w/ ____
4. do not form ____
- 10-15u
- chilling
- wet method: 70% alcohol
- ribbons
MOUNTING MEDIA REFRACTIVE INDEX:
AQUEOUS MEDIA
1. Water
2. Glycerin
3. Glycerin Jelly
- aka
4. Farrant’s medium
5. Apathy’s medium
- 1.33
- 1.46
- 1.47
- Kaiser’s 1880 - 1.43
- 1.52
MOUNTING MEDIA REFRACTIVE INDEX:
RESINOUS MEDIA
1. Canada Balsam
2. DPX
- aka
3. XAM
4. Clarite
- 1.524
- 1.532
- dibutylphthlate polystyrene xylene - 1.52
- 1.544
