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Question Answer
take out waterdehydrating
replace alcohol w/a reagent miscible w/parafinclearing
impregnation, permeationinfiltration
infiltration w/a supporting mediumembedding
chem reagent that can be used for dehydrating and clearing steps in tissue processor.universal solvent
volatile oils have odor of plant from which they are madeessential oil
water soluble (polyethylene glycols)carbowax
substance that changes one ion for another. used in decalcification and exchange an ammonium ion for the calcium ion removed from tissueion exchange
absorbs water from air so use it over a drying chemical like drierite or an anhydrous calcium chlorieeacetone
removing calcium from bonedecalcification
must be mixable with both dehydrating agent and infiltration mediumclearing agents
organic compounds that have property of binding certain metals,chelating agent
capable of mixing or being mixedmiscible
alcohol, dioxane, acetone, tetrahydrofurandehydrating agents
xylene, toluene, benzene, acetone, chloroform, cedarwood oil, tetrhydrofuran, dioxane, limonene derivatives, aliphatic hydrocarbonsclearing agents
infiltration mediumparaffin
dealcoholization agentsclearing agents
inadequate clearinginadequate infiltration
soft, mushy tissueinadequate clearing, inadequate infiltration results in
hard, brittle tissueprolonged period in clearing agent
most widely used clearing agent, used in processing and stainingxylene
prolonged treatment w/xylene during processingtissue becomes overhardened
rapid in displacing alcohol, mixiable w/paraffinxylene
turns cloudy in presence of water xylene
has PEL of 100ppm, use w/adequate ventilationxylene
doesnt overharden as much as xylenetoluene
best aromatic clearing agenttoluene
fast acting, doesnt overharden like xylenebenzene
hardens muscle,tendon and uterus more than toluenebenzene
evaporates rapidly from paraffin bathbenzene
when using this clearing agent, the paraffin used for infiltration does not require rotaion and changing as frequently as w/o clearing agentsbenzene
carcinoge, affects blood and bone marrowbenzene
leaves tissue less brittle than xylenechloroform
penetrates slowly, makes clearing a longer process chloroform
use in tightly covered containerschloroform
desiccates (dries) connective tissuechloroform
very volatile, makes fluid levels difficult to maintain in open processorchloroform and benzene
causes formation of phosgeneheating chloroform
a toxic gasphosgene
PEL 50ppm, it is carcinogenicchloroform
low boiling point 58C, will boil off and be replaced by paraffinacetone
can also be used for dehydrationacetone
Universal solventacetone
show more shrinkage than those cleared with xylenetissues cleared w/acetone
are very volatileessential oils
volality is not sufficient to allow ready replacment during paraffing infiltrationessential oils
if oil remains in tissue, microtomy is difficultessential oils
remove w/an aromatic hydrocarbon clearing agentessential oils
best known most widely used of the oilscedarwood oil
clears alcohol dehydrated tissue quickly and doesnt cause further shrinkagecedarwood oil
hardens and damages tissue less than and other known clearing agentcedarwood oil
tissue may remain in this oil indefinitelycedarwood oil
are expensive reagents and are not used routinely essential oils
have become popular during last dacade and have a strong citrus odorlimonene reagents
irritant, sensitizer in concetrated from, causes allergic reactionslimonene reagents
causes difficulty breathing, headacheslimonene reagents
PELS are unavailable for inhalation toxicitylimonene reagents
harden tissue less than xyelene but they cause more contamination of the paraffinlimonene reagents
when used in staining, these reagents are greasier limonene reagents
newest class of clearing agent aliphatic hydrocarbons (alkanes)
they are low in reactivity and toxicityaliphatic hydrocarbons (alkanes)
they are nonirritant and nonsensitizingaliphatic hydrocarbons (alkanes)
they have an 8hour exposure limit of 300ppmaliphatic hydrocarbons (alkanes)
used in histo, they penetrate tissue faster, remove fat effectively and allow coverslips to dry in usual mannerlight weight (short chains)aliphatics
they dont tolerate water and they are incompatible with some mounting mediadisadvantage of aliphatics
difficult to use in areas of high humidityaliphatic hydrocarbons
can be used on all tissue processors and automated stainersaliphatic hydrocarbons
not recommended for use on automatic coverslippersaliphatic hydrocarbons
dioxane, tertiary butanol, tetrahydrofuran, acetoneuniversal solvent (dehydrating and clearing)
adequate reagent volume must be maintained to ensure high quality tissue sections
have a schedule for the processor. rotate and change the reagentsto ensure consistant section quality
holds cells and intracellular structures, while thin sections are cutembedding media
most popular embedding media, large amounts of tissue can be processed in a short timeparaffin
is inert mixture of hydrocarbons produced by craking of petroleumparaffin
contains beeswax, rubber and other waxes or plasticscommercial paraffin
formed when the firs section cut adheres to microtome knife ribbons
paraffin becomes harder and provides better support for hard tissuemelting point increases
wax becomes soft and provides less suport for hard tissuemelting point decreases
should be matched to the hardness of tissue to be sectionedparaffin
enhance ability of paraffin compounds to provide support for hard tissuesadditives
55C to 58Cmelting point of paraffin
tissue should remain in it the shortest time necessary for good infiltrationparaffin
prolonged heat causes shrinkage and hardeningparaffin
keep supply at 2C to 4C above the melting pointparaffin supply
tissues exposed to overheated paraffin will overhaden, and section quality will be changed, tissue will not be effectedparaffin
must be recorded dailyparaffin temp
will become contaminated w/clearing agentm, if it is not changed and monitored frequentlyparaffin
three changes of paraffin are recommended for infiltration
aided by vacummparaffin infiltration
staining problemsoverprocessing
difficult to process on the same processing cycle should be processed overinght on an open processor w/o heat and vaccumbx, uterus, breast and large specimens
dehydratesalcohol
PEL 1,000 ppmEthyl alcohol
clear,colorless,flammableEthyl alcohol
drinkable, controlled by federal goverment, troublesome record keeping is required, reliable, fast acting, best dehydrantEthyl alcohol
is hydrophillicEthyl alcohol
mixable with water as well as organic solvents in all proportionsEthyl alcohol
should be used in sequence of solutions that gradually increase in concentrationEthyl alcohol
dehydration process is started at 95%, followed by absolute alcoholEhtyl alcohol
causes excessive shrinkage and hardeningEthyl alcohol
poisonous and rarely usedMethyl alcohol
flammable, unpleasant odor, clear, colorlessMethyl alcohol
PEL of 200Methyl alcohol
overexposure causes blindness and deathMethyl alcohol
substitute for ethanolisopropanol
no goverment restrictions. doesnt harden or shrink tissue as much as ethanolisopropanol
mildly irratant to eyes, nose, throatisopropanol
PEL of 400 ppmisopropanol
good dehydrant for plant and animalbutanol
pronounced odor and low dehydrating power.requires long period for dehydrationbutanol
excellent dehydrant for slow processingbutanol
causes less shrinkage and hardening than ethanolbutanol
rapid reacting, less expensive, causes excessive shrinkageacetone
PEL 1,000ppmacetone
flammable, flash point of -17cacetone
referred to as emebedding medium. holds cells and intracellular structures in proper relationship while thin sections are cutinfiltration media
Where the tissue cassettes are transported from one solution to nextopen system processor
no vacuum usedopen system processor
the tissue is stationary and fluids are pumped in and out of the closed chamber holding the tissue cassetteclosed system processor
uses vacuumclosed system processor
different protocols based on open or closed systemprocessor solutions and times
shorter processing cycle is used for bx specimens so they wont be overhardenedprocessing time
casting or blockingembedding
enclosing the tissue in the infiltration medium used for processing and then allowing the medium to solidifyembedding
is determined by the embedding medium usedmethod of processing
side down in the cassette is the side down in the moldembedding reminders
upside can be notched or inkedembedding reminders
tissue should be in center of moldembedding reminders
light pressure should be applied over the entire tissue during orientation and light chillingembedding reminders
wipe forceps inbetween samples to prevent forcep metastasisembedding reminders
place in a line parallel to the longer axis on the moldmultiple pieces
shuld be facing one side of the mold not up or downthe epithelium
embed on diagnol to aid in the ease of sectioningbone
GI tract, cysts and gall bladdertissue w/a wall
embed on an edge so all layers are showingGI tract, cysts and gall bladder (tissue w/a wall)
fallopian tubes, appendix and arteriesTubular structures
embed in a cross section so that the lume and all mucosa, submucosa and external muscle layers are obviousfallopian tubes, appendix and arteries (tubular structures)
micible w/ lower alcohols, water, ether, chloroform, acetone, benzene, toluene, xylene and melted paraffingtetrahydrofuran
same as dioxane in use but less expensivetetrahydrofuran
acts rapidly w/o causing excessive shrinkage and hardeningtetrahydrofuran
can be used to dehydrate and clear stained slidestetrahydrofuran
best universal solventtetrahydrofuran
very volatile, offensive odor, explosive peroxides may form, can cause conjuntivities, use in well ventilated roomtetrahydrofuran
produces less shrinkage than ethanoldioxane
faster dehydrant than ethanol but m/b used in larger volumesdioxane
can be reused by treating anhydrous calcium chloride or calcium oxidedioxane
it has water in it, so if itis left in the tissue, it may shrink it up to 50% during infiltrationdioxane
cummulatively toxic, pronounced odor, rarely useddioxane
Osha PEL 100ppm, NIOSH PEL 1ppm, carcinogen, flammabledioxane
expensive, tends to solidify at room temptertieary butanol
most satisfacory method of obtaining F.s. from formalin fxd tissue30% sucrose
micible w/ lower alcohols, water, ether, chloroform, acetone, benzene, toluene, xylene and melted paraffingtetrahydrofuran
same as dioxane in use but less expensivetetrahydrofuran
acts rapidly w/o causing excessive shrinkage and hardeningtetrahydrofuran
can be used to dehydrate and clear stained slidestetrahydrofuran
best universal solventtetrahydrofuran
very volatile, offensive odor, explosive peroxides may form, can cause conjuntivities, use in well ventilated roomtetrahydrofuran
produces less shrinkage than ethanoldioxane
faster dehydrant than ethanol but m/b used in larger volumesdioxane
can be reused by treating anhydrous calcium chloride or calcium oxidedioxane
it has water in it, so if itis left in the tissue, it may shrink it up to 50% during infiltrationdioxane
cummulatively toxic, pronounced odor, rarely useddioxane
Osha PEL 100ppm, NIOSH PEL 1ppm, carcinogen, flammabledioxane
expensive, tends to solidify at room temptertieary butanol
inital paraffin infiltration must be half tertiary butanol and half paraffintertiary butanol
can be used as a dehydrant in the staining processtertiary butanol
odorus, PEL 100ppmtertiary butanol
Dealcoholizationclearing
the removal of alcohol, to make tissue receptive to the infiltration medium clearing
used for processing of tissue and staining of microscopic sectionsclearing
hard brittle tissue, sectioning difficultexcessive clearing
reduces crystal size and increases stickiness and adhesionbeeswax
reduces brittleness, increases stickiness and maes the formation of ribbons during sectioningrubber
other waxes produce smooth texture and smaller crystal sizeParaffin composition
beeswax,rubber and other waxesParaffin composition
increases the hardness and supportplastics
infiltrate tissue directly from aqueous fixativewater soluble waxes
will not infiltrate tissue containing large amounts of fatcarbowax
cryoprotectant30% sucrose