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350 > Microtomy > Flashcards

Microtomy Flashcards

1
Q

how does a rotary microtome work

A

moving the block over a stationary blade and advancing the block each cycle

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2
Q

why is tissue cut 4-5um

A

single layer of cells

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3
Q

what tissues are cut 2-3um

A

lymph and kidney

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4
Q

what tissues are cut 6-8um

A

breast and brain

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5
Q

what are disposable blades not sharp enough to cut. what is used instead?

A

resin embedded blocks use glass or diamond to cut <1um

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6
Q

What is facing

A

trimming away excess wax to expose the full face of tissue

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7
Q

how long does the block have to cool for

A

at least 15 minutes

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8
Q

what is levelling

A

section is made, then trim 60-100 um and another section is made

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9
Q

what is serial sections

A

block is faced and every ribbon is put on slide without skipping sections

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10
Q

what are deepers or recuts from a pathologist

A

levels are made

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11
Q

what steps for microtomy

A
  1. locate safety mechanism
  2. adjust microtome setting
  3. ensure all levers are tight
  4. patient soaking the block
  5. short ribbons are fine
  6. keep it clean
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12
Q

when should the handwheel be locked

A

inserting/removing a block and not actively cutting

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13
Q

what are the microns for facing

A

20-30

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14
Q

what steps are done prior to fine cutting

A

fixaation
processing
microtome
temperature of block is softened

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15
Q

what steps happen during fine cutting

A

cutting speed = 1 rotation per second

ribbons

water bath = 10 degrees below MP of wax

section retrieval

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16
Q

Causes of curved sections

A

block edge not parallel
dull blade
ununiform tissue density

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17
Q

solution to curved sections

A

face until parallel
new blade

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18
Q

causes of chatter

A

loose lever vibration
high clearance angle
hard tissue
OVERDEHYDRATED tissue
dull blade

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19
Q

solutions to chatter

A

tighten levels
lower clearance angle
surface decal/ rehydrate tissue
new blade

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20
Q

causes of holes in section

A

facing aggressively
hard particles in the tissue

21
Q

solution of holes in section

A

lower micrometer setting
soak block in water

22
Q

section splitting perpendicular to blade

A

nick in blade
calcification
hard particles

23
Q

solution to splitting perpendicular to blade

A

new blade
surface decal
reembed

24
Q

fail to form ribbons causes

A

too hard wax
wrong clearance angle

25
Q

solution to failing form of ribbons

A

warm the block with thumb
increase speed
adjust clearance angle

26
Q

sections attach to block on return stroke causes

A

low clearance angle
debris
microtome retraction now working
STATIC ELECTRICITY

27
Q

solution to section attaching to block

A

increase clearance angle
HUMIDIFY the room

28
Q

causes of incomplete sections

A

not fully faced
incomplete paraffin infiltration

29
Q

solution to incomplete sections

A

reface block
reembed or reprocess

30
Q

causes of compression

A

dull blade
soft or warm paraffin

31
Q

solution to compression

A

new blade
cool block
reembed in harder wax

32
Q

causes of expanding section in waterbath

A

incomplete tissue
high water bath temp

33
Q

soluition to expansion in waterbath

A

reprocess
decrease water bath temp

34
Q

causes of section rolling into cylinder instead of flat

A

dull blade
too thick sections

35
Q

solution to rolling sections

A

new blade
thinner sections

36
Q

what temperature is the water bath

A

41-45 degrees

or

10 degrees lower than MP of wax

37
Q

too cold waterbath causes what to sections

A

wrinkles not come out

38
Q

warm waterbath will cause what to section

A

melt and disintegrate section

39
Q

how long and temperature of the oven

A

30 min - 1 hr at <60C

usually 37C

40
Q

what is a disadvantage of using microwaves to dry slides

A

cooking the water and fat of the tissue

41
Q

purpose of the oven

A

remove any water from section

NOT MELT WAX

42
Q

what happens if water is on the slides when staining

A

deparrifinization is incomplete

43
Q

when are adhesive or charged slides used

A

when slides undergo harsh solutions or pretreatments during staining

eg silver stains or IHC (moisture and enzymes)

44
Q

what are common adhesives

A

gelatin
albumin
elmers glue
poly L lysine

45
Q

disadvantage of adhesives

A

produce background saining

46
Q

T/F charge slides are positively charged

A

T

47
Q

what method to make charge slides

A

3-aminopropyltriethoxysilane (APTES)

48
Q

how does APTES work

A

covalent bind to glass making an amino group coating

49
Q

causes of parched earth

A

improper processing
tissue chilled too rapidly (cryosprays)
floating on waterbath too long

350 (7 decks)

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