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Processing

Quiz yourself by thinking what should be in each of the black spaces below before clicking on it to display the answer.
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Question
Answer
take out water   dehydrating  
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replace alcohol w/a reagent miscible w/parafin   clearing  
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impregnation, permeation   infiltration  
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infiltration w/a supporting medium   embedding  
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chem reagent that can be used for dehydrating and clearing steps in tissue processor.   universal solvent  
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volatile oils have odor of plant from which they are made   essential oil  
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water soluble (polyethylene glycols)   carbowax  
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substance that changes one ion for another. used in decalcification and exchange an ammonium ion for the calcium ion removed from tissue   ion exchange  
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absorbs water from air so use it over a drying chemical like drierite or an anhydrous calcium chloriee   acetone  
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removing calcium from bone   decalcification  
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must be mixable with both dehydrating agent and infiltration medium   clearing agents  
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organic compounds that have property of binding certain metals,   chelating agent  
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capable of mixing or being mixed   miscible  
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alcohol, dioxane, acetone, tetrahydrofuran   dehydrating agents  
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xylene, toluene, benzene, acetone, chloroform, cedarwood oil, tetrhydrofuran, dioxane, limonene derivatives, aliphatic hydrocarbons   clearing agents  
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infiltration medium   paraffin  
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dealcoholization agents   clearing agents  
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inadequate clearing   inadequate infiltration  
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soft, mushy tissue   inadequate clearing, inadequate infiltration results in  
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hard, brittle tissue   prolonged period in clearing agent  
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most widely used clearing agent, used in processing and staining   xylene  
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prolonged treatment w/xylene during processing   tissue becomes overhardened  
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rapid in displacing alcohol, mixiable w/paraffin   xylene  
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turns cloudy in presence of water   xylene  
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has PEL of 100ppm, use w/adequate ventilation   xylene  
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doesnt overharden as much as xylene   toluene  
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best aromatic clearing agent   toluene  
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fast acting, doesnt overharden like xylene   benzene  
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hardens muscle,tendon and uterus more than toluene   benzene  
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evaporates rapidly from paraffin bath   benzene  
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when using this clearing agent, the paraffin used for infiltration does not require rotaion and changing as frequently as w/o clearing agents   benzene  
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carcinoge, affects blood and bone marrow   benzene  
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leaves tissue less brittle than xylene   chloroform  
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penetrates slowly, makes clearing a longer process   chloroform  
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use in tightly covered containers   chloroform  
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desiccates (dries) connective tissue   chloroform  
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very volatile, makes fluid levels difficult to maintain in open processor   chloroform and benzene  
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causes formation of phosgene   heating chloroform  
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a toxic gas   phosgene  
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PEL 50ppm, it is carcinogenic   chloroform  
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low boiling point 58C, will boil off and be replaced by paraffin   acetone  
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can also be used for dehydration   acetone  
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Universal solvent   acetone  
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show more shrinkage than those cleared with xylene   tissues cleared w/acetone  
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are very volatile   essential oils  
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volality is not sufficient to allow ready replacment during paraffing infiltration   essential oils  
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if oil remains in tissue, microtomy is difficult   essential oils  
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remove w/an aromatic hydrocarbon clearing agent   essential oils  
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best known most widely used of the oils   cedarwood oil  
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clears alcohol dehydrated tissue quickly and doesnt cause further shrinkage   cedarwood oil  
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hardens and damages tissue less than and other known clearing agent   cedarwood oil  
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tissue may remain in this oil indefinitely   cedarwood oil  
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are expensive reagents and are not used routinely   essential oils  
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have become popular during last dacade and have a strong citrus odor   limonene reagents  
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irritant, sensitizer in concetrated from, causes allergic reactions   limonene reagents  
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causes difficulty breathing, headaches   limonene reagents  
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PELS are unavailable for inhalation toxicity   limonene reagents  
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harden tissue less than xyelene but they cause more contamination of the paraffin   limonene reagents  
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when used in staining, these reagents are greasier   limonene reagents  
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newest class of clearing agent   aliphatic hydrocarbons (alkanes)  
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they are low in reactivity and toxicity   aliphatic hydrocarbons (alkanes)  
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they are nonirritant and nonsensitizing   aliphatic hydrocarbons (alkanes)  
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they have an 8hour exposure limit of 300ppm   aliphatic hydrocarbons (alkanes)  
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used in histo, they penetrate tissue faster, remove fat effectively and allow coverslips to dry in usual manner   light weight (short chains)aliphatics  
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they dont tolerate water and they are incompatible with some mounting media   disadvantage of aliphatics  
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difficult to use in areas of high humidity   aliphatic hydrocarbons  
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can be used on all tissue processors and automated stainers   aliphatic hydrocarbons  
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not recommended for use on automatic coverslippers   aliphatic hydrocarbons  
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dioxane, tertiary butanol, tetrahydrofuran, acetone   universal solvent (dehydrating and clearing)  
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adequate reagent volume must be maintained   to ensure high quality tissue sections  
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have a schedule for the processor. rotate and change the reagents   to ensure consistant section quality  
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holds cells and intracellular structures, while thin sections are cut   embedding media  
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most popular embedding media, large amounts of tissue can be processed in a short time   paraffin  
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is inert mixture of hydrocarbons produced by craking of petroleum   paraffin  
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contains beeswax, rubber and other waxes or plastics   commercial paraffin  
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formed when the firs section cut adheres to microtome knife   ribbons  
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paraffin becomes harder and provides better support for hard tissue   melting point increases  
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wax becomes soft and provides less suport for hard tissue   melting point decreases  
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should be matched to the hardness of tissue to be sectioned   paraffin  
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enhance ability of paraffin compounds to provide support for hard tissues   additives  
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55C to 58C   melting point of paraffin  
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tissue should remain in it the shortest time necessary for good infiltration   paraffin  
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prolonged heat causes shrinkage and hardening   paraffin  
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keep supply at 2C to 4C above the melting point   paraffin supply  
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tissues exposed to overheated paraffin will overhaden, and section quality will be changed, tissue will not be effected   paraffin  
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must be recorded daily   paraffin temp  
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will become contaminated w/clearing agentm, if it is not changed and monitored frequently   paraffin  
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three changes of paraffin are recommended   for infiltration  
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aided by vacumm   paraffin infiltration  
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staining problems   overprocessing  
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difficult to process on the same processing cycle should be processed overinght on an open processor w/o heat and vaccum   bx, uterus, breast and large specimens  
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dehydrates   alcohol  
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PEL 1,000 ppm   Ethyl alcohol  
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clear,colorless,flammable   Ethyl alcohol  
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drinkable, controlled by federal goverment, troublesome record keeping is required, reliable, fast acting, best dehydrant   Ethyl alcohol  
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is hydrophillic   Ethyl alcohol  
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mixable with water as well as organic solvents in all proportions   Ethyl alcohol  
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should be used in sequence of solutions that gradually increase in concentration   Ethyl alcohol  
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dehydration process is started at 95%, followed by absolute alcohol   Ehtyl alcohol  
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causes excessive shrinkage and hardening   Ethyl alcohol  
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poisonous and rarely used   Methyl alcohol  
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flammable, unpleasant odor, clear, colorless   Methyl alcohol  
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PEL of 200   Methyl alcohol  
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overexposure causes blindness and death   Methyl alcohol  
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substitute for ethanol   isopropanol  
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no goverment restrictions. doesnt harden or shrink tissue as much as ethanol   isopropanol  
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mildly irratant to eyes, nose, throat   isopropanol  
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PEL of 400 ppm   isopropanol  
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good dehydrant for plant and animal   butanol  
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pronounced odor and low dehydrating power.requires long period for dehydration   butanol  
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excellent dehydrant for slow processing   butanol  
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causes less shrinkage and hardening than ethanol   butanol  
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rapid reacting, less expensive, causes excessive shrinkage   acetone  
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PEL 1,000ppm   acetone  
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flammable, flash point of -17c   acetone  
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referred to as emebedding medium. holds cells and intracellular structures in proper relationship while thin sections are cut   infiltration media  
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Where the tissue cassettes are transported from one solution to next   open system processor  
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no vacuum used   open system processor  
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the tissue is stationary and fluids are pumped in and out of the closed chamber holding the tissue cassette   closed system processor  
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uses vacuum   closed system processor  
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different protocols based on open or closed system   processor solutions and times  
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shorter processing cycle is used for bx specimens so they wont be overhardened   processing time  
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casting or blocking   embedding  
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enclosing the tissue in the infiltration medium used for processing and then allowing the medium to solidify   embedding  
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is determined by the embedding medium used   method of processing  
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side down in the cassette is the side down in the mold   embedding reminders  
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upside can be notched or inked   embedding reminders  
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tissue should be in center of mold   embedding reminders  
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light pressure should be applied over the entire tissue during orientation and light chilling   embedding reminders  
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wipe forceps inbetween samples to prevent forcep metastasis   embedding reminders  
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place in a line parallel to the longer axis on the mold   multiple pieces  
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shuld be facing one side of the mold not up or down   the epithelium  
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embed on diagnol to aid in the ease of sectioning   bone  
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GI tract, cysts and gall bladder   tissue w/a wall  
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embed on an edge so all layers are showing   GI tract, cysts and gall bladder (tissue w/a wall)  
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fallopian tubes, appendix and arteries   Tubular structures  
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embed in a cross section so that the lume and all mucosa, submucosa and external muscle layers are obvious   fallopian tubes, appendix and arteries (tubular structures)  
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micible w/ lower alcohols, water, ether, chloroform, acetone, benzene, toluene, xylene and melted paraffing   tetrahydrofuran  
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same as dioxane in use but less expensive   tetrahydrofuran  
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acts rapidly w/o causing excessive shrinkage and hardening   tetrahydrofuran  
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can be used to dehydrate and clear stained slides   tetrahydrofuran  
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best universal solvent   tetrahydrofuran  
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very volatile, offensive odor, explosive peroxides may form, can cause conjuntivities, use in well ventilated room   tetrahydrofuran  
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produces less shrinkage than ethanol   dioxane  
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faster dehydrant than ethanol but m/b used in larger volumes   dioxane  
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can be reused by treating anhydrous calcium chloride or calcium oxide   dioxane  
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it has water in it, so if itis left in the tissue, it may shrink it up to 50% during infiltration   dioxane  
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cummulatively toxic, pronounced odor, rarely used   dioxane  
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Osha PEL 100ppm, NIOSH PEL 1ppm, carcinogen, flammable   dioxane  
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expensive, tends to solidify at room temp   tertieary butanol  
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most satisfacory method of obtaining F.s. from formalin fxd tissue   30% sucrose  
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micible w/ lower alcohols, water, ether, chloroform, acetone, benzene, toluene, xylene and melted paraffing   tetrahydrofuran  
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same as dioxane in use but less expensive   tetrahydrofuran  
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acts rapidly w/o causing excessive shrinkage and hardening   tetrahydrofuran  
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can be used to dehydrate and clear stained slides   tetrahydrofuran  
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best universal solvent   tetrahydrofuran  
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very volatile, offensive odor, explosive peroxides may form, can cause conjuntivities, use in well ventilated room   tetrahydrofuran  
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produces less shrinkage than ethanol   dioxane  
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faster dehydrant than ethanol but m/b used in larger volumes   dioxane  
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can be reused by treating anhydrous calcium chloride or calcium oxide   dioxane  
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it has water in it, so if itis left in the tissue, it may shrink it up to 50% during infiltration   dioxane  
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cummulatively toxic, pronounced odor, rarely used   dioxane  
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Osha PEL 100ppm, NIOSH PEL 1ppm, carcinogen, flammable   dioxane  
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expensive, tends to solidify at room temp   tertiary butanol  
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inital paraffin infiltration must be half ___ _____ and half paraffin   tertiary butanol  
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can be used as a dehydrant in the staining process   tertiary butanol  
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odorus, PEL 100ppm   tertiary butanol  
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Dealcoholization   clearing  
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the removal of alcohol, to make tissue receptive to the infiltration medium   clearing  
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used for processing of tissue and staining of microscopic sections   clearing  
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hard brittle tissue, sectioning difficult   excessive clearing  
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reduces crystal size and increases stickiness and adhesion   beeswax  
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reduces brittleness, increases stickiness and maes the formation of ribbons during sectioning   rubber  
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other waxes produce smooth texture and smaller crystal size   Paraffin composition  
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beeswax,rubber and other waxes   Paraffin composition  
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increases the hardness and support   plastics  
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infiltrate tissue directly from aqueous fixative   water soluble waxes  
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will not infiltrate tissue containing large amounts of fat   carbowax  
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cryoprotectant   30% sucrose  
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