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MIDTERMS HISTOPATH LAB > Microtomy > Flashcards

Microtomy Flashcards

1
Q

______: previously processed tissue, is trimmed and cut into uniformly thin slices or sections
1. aka

A

SECTIONING

  1. microtomy
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2
Q

3 BASIC PARTS
1. where the embedded tissue is held in position
2. perform the actual cutting of tissue sections
3. line up tissue block in
proper position with the knife, adjust proper thickness

A
  1. BLOCK HOLDER
  2. KNIFE CARRIER AND KNIFE
  3. PAWL, RATCHET FEED WHEEL AND ADJUSTMENT SCREWS
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3
Q

General Principle of Microtomy

A

A spring-balanced teeth (pawl) is brought in contact with, and turns a ratchet feed wheel connected to a micrometer screw, which is in turn rotated, moving the tissue block at a predetermined distance towards the knife for cutting sections at uniform thickness

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4
Q

TYPES OF MICROTOME:

  1. for small and large paraffin-embedded blocks; not for serial sections
    - aka
    - thickness
    - inventor
  2. for paraffin embedded blocks (routine procedure); up and down vertical movement
    - aka
    - thickness
    - inventor
A
  1. ROCKING
    - CAMBRIDGE
    - 10-12um
    - PALDWELL TREFALL
  2. ROTARY
    - MINOT
    - 4-6um
    - MINOT
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5
Q

TYPES OF MICROTOME:

  1. most dangerous
    - thickness
    - inventor
    - movable exposed knife that’s why it is the most dangerous which is also for celloidin
    - not movable knife; hard and tough tissue blocks in all forms of media
  2. for electron microscopy
    - thickness
    - small specimen fixed in ____ and embedded in ____
  3. undehydrated tissues in frozen state
    - thickness
    - inventor
    - ___ & ____ tissue constituents
A
  1. SLIDING
    - 4-9um
    - Adams
    - Standard Sliding
    - Base-Sledge
  2. Ultrathin
    - 0.5um
    - osmium tetroxide; plastic
  3. Freezing
    - 10-15um
    - Queckett
    - fats and heat sensitive
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6
Q

_____: rotary microtome inside a cold chamber

  1. temp is maintained between __ to __ (ave. is __)
  2. optimum temp
A

COLD MICROTOME/CRYOSTAT

  1. -5 to -30C (ave. is -20C)
  2. -18 to -20C
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7
Q

COLD MICROTOME/CRYOSTAT

  1. -5 to -15C
    - organs: (6)
  2. -15 to -25C
    - organs:(4)
  3. -35C
    - organs: (2)
A
  1. -5 to -15C
    - BRAIN, LYMPH NODES, LIVER, SPLEEN, UTERINE, CURETTAGE, SOFT TUMORS
  2. -15 to -25C
    - NON FATTY BREAST, PROSTATE, TONGUE, GI TRACT
  3. -35C
    - FATTY BREAST, OMENTAL TISSUE
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8
Q

FREEZING MICROTOME:

  1. CO2
    - knife
    - tissue
    - environment
  2. _____: warm tissue surface -> ____
A
  1. CO2
    - -40 to -60
    - -5 to -10
    - 0 to -10
  2. Decoline; thaw
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9
Q

FREEZING FRESH UNFIXED TISSUE
1. liquified nitrogen
2. isopentane cooled by liquified nitrogen
3. dry ice
4. carbon dioxide
5. aerosol spray

A
  1. -190
  2. -150
  3. -70
  4. -70
  5. -50
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10
Q

FREEZING PREVIOUSLY FIXED TISSUE

  1. FORMALIN FIXED TISSUE
    - coat slide w/ ______ or ____ to attach fixed tissues to the slide
    - immerse tissue block in boiling _____ for ____ before freezing & sectioning
    - histochemistry & lipids: ______ @ ____
  2. ALCOHOL FIXED TISSUE
    - wash in ____ form____ before sectioning
A
  1. FORMALIN FIXED TISSUE
    - ALBUMIN or CHROME-GLYCERIN JELLY
    - 10% BUFFERED FORMALIN; 1-2mins
    - 10% FORMOL CALCIUM;;4C
  2. ALCOHOL FIXED TISSUE
    - WATER; 12-24hrs
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11
Q

KINDS MICROTOME KNIVES:

Light Microscopy
1. for frozen sections; extremely hard and tough specimens in paraffin
- length
2. for paraffin section
- length
3. Plane-Concave
- length:
- plane side:
- concave side:

A

Light Microscopy
1. Plane-Wedge
- 100mm
2. Biconcave
- 120mm
3. Plane-Concave
- 25mm
- celloidin
- paraffin

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12
Q

KINDS MICROTOME KNIVES:

Electron Microscopy
1. resin blocks
2. trimming; semi thin sectioning

Others
1. sharp cutting edge capable of producing ____ sections
2. partially calcified tissues, paraffin, and frozen sections
- ___ and above

A

Electron Microscopy
1. Diamond-edge
2. Broken Glass Knives

Others
1. Disposable blades; 2-4um
2. Safety razor blades
- 10um and above

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13
Q

ANGLES DURING SECTIONING

  1. between the edge of the knife & the tissue block
    - degrees
  2. angle of cutting
    - degrees
  3. angle of cutting facet
    - degrees
A
  1. Clearance angle
    - 5-10 degrees
  2. Wedge angle
    - 15 degrees
  3. Bevel angle
    - 27-32 degrees
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14
Q

SHARPENING OF MICROTOME KNIVES

the cutting edge must be of good quality steel:
1. doesn’t maintain the edge
2. likely to nick against hard objects

tests in order to assess the sharpness of the cutting edge:
1. should cut a paraffin wax block at _____ thickness w/o serrations when examined under the microscope (100 X)
2. ____ criterion

A

the cutting edge must be of good quality steel:
1. TOO SOFT
2. TOO HARD

tests in order to assess the sharpness of the cutting edge:
1. 2-4um
2. VON MHOL’S CRITERION

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15
Q

______: Removal of gross nicks and blemishes

  1. material
    - best result
    - more polishing effect
    - coarsest
  2. lubricant
  3. direction
  4. no of strokes
    - ____ in each direction
    - ____ each surface (minot and plane wedge knives)
A

HONING

  1. Smooth stones
    - Belgium yellow
    - Arkansas
    - Fine carborundum
  2. Soapy water, Oil, Xylene, Liquid Paraffin
  3. Heel-to-Toe, edge first
  4. no of strokes
    - 20-30 strokes in each direction
    - 10-20 strokes each surface (minot and plane wedge knive
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16
Q

______: Removal of burrs, for final polishing and
sharpening of the cutting edge

  1. material
  2. lubricant
  3. direction
  4. no of strokes
A

STROPPING

  1. Horse leather
  2. Vegetable Oil
  3. Toe-to-Heel, edge last
  4. 40-120 double strokes
17
Q

OTHER HONING PROCEDURES

  1. Plate Glass Honing
    - _____ used for final polishing
  2. after repeated sharpening
    - widened bevel angle
  3. _____
A
  1. Plate Glass Honing
    - Diamantine
  2. Factory Grinding
    - >35 degreew
  3. Automatic hones
18
Q

PROPER CARE OF THE MICROTOME KNIFE

  1. Always use a back, when required, for sharpening. This is not necessary with ______ knives
  2. Knife should be cleaned with. ___ or ___ before and after use
  3. clean paraffin debris and small pieces of tissues with. ____
  4. cutting edge must be ____ & ____
  5. cutting edge must be ____ from _____ to be cut
A
  1. bi concave knives
  2. xylene or toluene
  3. soft brush
  4. sharp and smooth
  5. thinner from section

MIDTERMS HISTOPATH LAB (12 decks)

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