HT 12 Exam 1
Quiz yourself by thinking what should be in
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surgery | show 🗑
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show | the tissue is described and dissected, looking for abnormalities
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show | stabilization of proteins to prevent autolysis and putrefaction
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processing | show 🗑
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embedding | show 🗑
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microtomy | show 🗑
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show | a mounting medium
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show | a. Stabilization of proteins to prevent autolysis and putrefaction
b. Maintain proper relationship between cells and extracellular material
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show | To embed the tissue in a solid medium firm enough to support the tissue and give it sufficient rigidity to enable thin sections to be cut, and yet soft enough no to damage the knife or tissue
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Define and discuss dehydration | show 🗑
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show | to remove the dehydrating fluid from the tissue
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show | Maintains structure of tissue for cutting
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show | Fixing the tissue specimen firmly into the embedding medium, allowing it to harden-->ready to cut
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show | a. The sectioning of tissue for examination by microscopy
b. Sections routinely cut at 4-6 microns (um)
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Discuss staining as it applies to Histotechnology | show 🗑
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show | Protection and for better viewing under the microscope
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Pathology | show 🗑
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Histology | show 🗑
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Cytology | show 🗑
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Histologist | show 🗑
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show | a technical histology concerned especially with preparing and processing histological specimens
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show | a technician who specializes in histotechnology
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show | removal of living tissue for examination under the microscope
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Autolysis | show 🗑
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show | rotting degradations from micro organs
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show | an examination of a dead body to determine the cause of death
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Miscible | show 🗑
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Microtome | show 🗑
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show | Orientation of the tissue in infiltration medium and allowing it to solidify. Also known as casting or blocking. Areas of interest should be embedded down.
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show | Wall tissues must be embedded on edge (gallbladder, intestine, skin)
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show | tubular structures (with a LUMEN) are embedded in cross section (on end)
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Describe proper specimen orientation when embedding bone. | show 🗑
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Describe double embedding | show 🗑
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show | Recording number of pieces and blocks submitted, plus any special instructions (stains, embedding instructions). Only embed one cassette at a time, ensure forceps are clean between embedding different tissues.
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Define longitudinal section | show 🗑
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show | a cut that is perpendicular to the longest dimension
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Define oblique section | show 🗑
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show | a cut that only touches or grazes the surface
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show | a cut that divides into left and right
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Define lumen | show 🗑
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show | Clean the forceps between embedding of different tissues to prevent cross contamination. Areas of interest should be tampered so that the tissues lays flat down ensuring the entire specimen is cut. Cooling the blocks as rapidly as possible.
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show | Tampers are used to flatten tissue in the mold. Base molds form the shape of the block to be sectioned (reusable, disposable, and peel away). Cassettes hold and identify the specimen to be embedded.
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show | Area of interest can be marked with a notch or ink. Some facilities use "ink side up" while others use "ink side down."
India ink, Davidson Marking, tissue dyes, silver nitrate, and tattoo ink.
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show | 2-4 C above melting point for the paraffin.
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show | embedded tissue passes over the knife edge cutting the section.
each turn advances the number of micrometers.
used in cryostats.
(most common)
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show | block is held stationary and the knife travels over the specimen.
block advances up to the knife at a preset distance.
usually used for larger blocks in research
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freezing microtome | show 🗑
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show | sample is placed into a fixed holder, which then moves backwards and forwards across a knife
cuts sections of plastic, wood, and other hard materials
cuts sections at 1-60 um
biconcave knives
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show | ultra thin sections of specimens are cut.
uses glass/diamond knives.
0.5-2 um
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rocking microtome | show 🗑
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show | accurately cut tissue without freezing or embedding.
cutting using a vibrating blade.
used for difficult biological samples.
30-50 um for live tissue.
10-500 um for fixed tissue.
maintains cell morphology.
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steel knives | show 🗑
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show | most common.
superior edges.
refined, thickened razor blades.
high and low profile.
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tungsten carbide | show 🗑
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show | used for ultramicrotomy. hard but brittle.
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show | expensive but extremely durable due to hardness. used for ultramicrotomy
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show | concave on both sides.
used on rocking or sledge type microtomes.
rarely used in U.S.
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planoconcave | show 🗑
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plano-wedge (wedge) | show 🗑
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chatter | show 🗑
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show | all consecutive sections from a block are saved
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show | block is trimmed, sections are cut and picked up, the more of the block is trimmed away, and more sections are cut and picked up
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show | using a blade to manually trim away excess material
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washboarding | show 🗑
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clearance angle | show 🗑
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microtome saftey | show 🗑
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show | taking breaks
using an automated rotary microtome
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show | cause: knife too dull or dirty
remedy: sharpen/replace blade
cause: too little knife tilt
remedy: increase knife angle
cause: block too warm
remedy: chill block
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identify problem and the appropriate corrective action: split ribbons | show 🗑
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identify problem and the appropriate corrective action: length wise scratches | show 🗑
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identify problem and the appropriate corrective action: gouging (overlapping) | show 🗑
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show | cause: horizontal edges of block not parallel
remedy: trim block until parallel, reposition specimen holder
cause: blade blunt in one area
remedy: move blade or replace
cause: tissue varies in consistency
remedy: rotate block
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show | cause: over dehydration
remedy: rehydrate
cause: too much knife tilt
remedy: adjust knife angle
cause: dull knife
remedy: sharpen knife or replace blade
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identify problem and the appropriate corrective action: lifting sections | show 🗑
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identify problem and the appropriate corrective action: udulations/washboarding | show 🗑
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identify problem and the appropriate corrective action: holes in sections | show 🗑
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identify problem and the appropriate corrective action: lack of ribbon forming | show 🗑
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show | cause: block or blade loose
remedy: tighten
cause: too little knife tilt
remedy: increase clearance angle
cause: wax too soft
remedy: cool block or change to higher melting point wax
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show | whether fast or slow a constant speed is necessary to avoid uneven sections
one revolution of the hand-wheel per second
smooth even revolutions
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show | breathe on the block to provide heat, moisture, reduce static.
softening paraffin embedded tissue by trimming the block until the tissue is exposed and then soaking the block in H2O or glycerol.
tissues containing keratin can be soften by a depilatory
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describe routine mircrotome maintenance | show 🗑
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Optimal temperature for the flotation bath? | show 🗑
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show | deoionized water prevents static and contamination
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show | Alcohol, detergent, photo-flo can be added to the flotation bath to lower the surface tension.
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Identify the need for drying slides | show 🗑
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