UND 362 DC and Gross
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| What general info is given during grossing | ID, fresh v FS, diagrams and margins, pin out specs, remove clips, wrap small specs
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| what is the typical thickness/dimension that a spec should be when grossed in? | 3mm thick with 2mm buffer w/in cassette
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| what are the special prep specs in grossing | FS, micro, photographs, E/M with fix of gluteraldehyde or paraformadehyde, chromosome analysis, calculi, hormone receptors
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| what are the rush specs in grossing | endoscopic bx's, transplant tissues, transplant rejection tissues
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| what is the typical fixative in grossing and 3 special ones | formalin, bouins for trichrome, zenkers for PTAH, and B5 for bone marrow
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| What can be done with fatty tissue to clear | use acetone, but it will alter nuclear morphology
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| what should be avoided with decal pertaining to bone | avoid bone with soft tissue
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| what is ink used for in grossing | orient surface, margin, sides; proximal and distal margins;all margins on breast inkend
darker colers work better with microscope
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| what should be avoided when inking | areas containing an lesion
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| how many measurements are taken during gross | 2 or 3 dimensions (or aggregate), smallest to largest diameter
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| what are some of the histology terms used in grossing pertaining to sections? | cross section, transverse section, longitudinal section, on edge, on end, en face, cut side down
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| Pertaining to legal issues in histology name a couple socially sensitive and a likely forensics spec (and what must be done with it) | socially (POC, abortions)
forensics - bullets, must follow chain of custody and MUST NOT be let unattended
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| how are skins cut | Punchs are bisected larger than .5cm dia
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| Lymph nodes are typically for? and for what specs | FS, micro, Touch prep, flow, cytogenetics, hormones, biochem assay (mostly from breast,prostate, colon
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| what is the proper dimensions bone | 3-5mm (not less than 2 or it will release from paraffin)
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| dense bone v porous will take what timeframe | dense bone will take longer than porous
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| what is the typical fixative for bone | formalin (not recommended to use alcoholic formalin or 70% alcohol because it will slow the process)
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| what is the positive and negative for using zenkers with bone marrow | dual purpose fix and decal, but it can dissolve iron and contains mercury (mercury pigment). it will also cause radiopacity.
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| what is the best way to set the cassettes in decal | suspend or set on gauze (do not place flat on botton, all sides should be exposed)
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| what must be done with decal'd bone before putting it in formalin? | MUST be washed or the decal (hcl) and formalin will cause bis-chloromethyl ether
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| what fixative can be used for bone and what must it be placed in afterwards | picric acid (but must go directly into alcohol)
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| name 4 decal agent properties | speed, prevent cell damage (via acid), preserve tissue morphology, change stain ability of tissue
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| List 4 decal methods | Acid, chelating, ion-exchange, electrical ionization (microwave)
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| what happens during acid decalcification | calcuim salts (carbonate and phosphate) dissolve and then ionize (ie gain pos. or neg. charge) when exposed to acids
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| at what ph is calcium soluble at | 4.5
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| name the 2 strong acids for decal and 4 properties | nitric or HCL
INORganic, rapid, recommended conc. 5-10%, affects stainibility (time must be monitored)
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| name 3 weak acids and give 2 properties | formic, acetic, picric
ORGanic, slow but gentle
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| what is the proper decal method | wash-dc-check dc (chemically or via bone), wash, if not full dc replace solution and repeat until done, WASH
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| what are four common descripters for using formic acid as a DC? | its common, slow, weak, organic
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| name the 6 formic acid decalcifiers | lilly, evans & krajian, Kristensens, Gooding and Stewart, Commercial (eg immunocal), and one with no name
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| what is the formula for lilly DC | 5% formic acid (works ok at room temp but at 37○C (98○F) it will interfere with staining)
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| what is the formala for evans and krajian DC | 25% formic acid and 10% sodium citrate
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| what is the formula for Kristensens DC | sodium formate and formic acid
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| what is the formula for gooding and steward DC | formic acid and formalin
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| what is a formic DC formula that doesn't have a trademark | 8% formic acid and 8% HCL
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| How are HCL and Nitric Acid DC's in terms of speed | They are rapid DC
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| Using HCL DC at 37○C will affect eosin, alum/weigerts hematox, feulgen and azure eiosin stains | Eosin will be OK, Alum/Weigerts will be impaired, Feulgen will not work, and azure eosin will give pink cytoplasm and nuclear stain
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| using HCL DC at 55○C will cause what | lost of Ca salt and hydrolysis of bone matrix
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| HCL DC will cause what and how can it be prevented | hcl will cause tissue swelling, it can be prevented by adding chromic acid, alcohol, or NACL to the HCL
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| List 3 HCL DC formulas and their formula | von ebers (Hcl/sodium chloride), richman-gelfand-hill (formic/HCL), commercial DC's containg hcl
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| give 4 facts about Nitric acid as a DC | rapid, best after formalin fixation, will impair nuclear staining after 48 hrs, will produce an unsatisfactory geimsa stain
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| What are the 4 Nitric acid DC formulas | 5-10% aqueous nitric acid, perenyis fluid (10% nitric acid, alcohol, chromic acid), 5% nitric acid/formalin, Commercial nitric DC
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| What is chelating DC | organic (binding w/ metals)
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| give 4 facts pertaining to chelating DC | slow but gentle, doesn't damage tissue or ability to stain (therefore good for EM and IHC/enzymes, takes 2-4 days for 3mm section, exact ph not critical
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| how does chelating work? | EDTA binds calcium ion from outer shell of apetite crystal (which gets reduced)
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| How does Ion exchange work for DC | ammoniom ions are exchanged for Ca ions (formic acid remains clear of Ca
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| Describe visually what an ion exchange setup would look like | glass jar with 10-20% resin (ammoniated salt) on the bottom and 80-90%formic acid on top (with bone laying on top of resin
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| what are the advantages of using ion exchange resin for DC | good preservation, eliminates daily changes, resin can be reclaimed
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| describe using electrical ionization for DC | (rarely used), elect. current to solution, the current draws the neg. calcuim to the + carbon cathode
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| what are two disadvantages to using electrical ionization | heat is generated therefore potential damage, limited specs can be done at a time
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| what are 3 positives and 1 possible drawback to using a microwave for DC | can fix and DC, reduced time, consistent slides
be careful not to OVER decalcify
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| give 7 factors affecting DC rate | conc. and volume, temp, agitation, suspension, type of bone, spec size, patients age.
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| how does concentration and volume affect dc? | increased conc. will decrease the DC time but will increase staining problems. (use large volumes for multiple specs)
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| how does temp affect DC | increase temp will decrease time but will increase staining problems (it is best at 20-25○C)
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| what are the two types of bone and how would they affect DC | cortical (hard), cancellious (spongy) bone. Harder bone would take longer
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| what are the 5 ways that the DC endpoint can be determined | xray (most accurate), chemical (accurate), physical (possible damage), weight loss/gain, bubble test
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| how can you use a chemical to check for dc end pt. | take 5ml of each (dc fluid, 5% ammonium hydroxide, 5% ammonium oxalate) check for turbidity (if precipitate then DC not complete, change DC)
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| what are 4 facts about using xray to check for dc end pt | uses visible evidence (pre and then post xray), expensive, BEST METHOD, can't be used with zenkers or B5 fixed bone because they have mercuric chlorid which will cause radiopacity
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| how is DC checked physically | done by probing or bending (not recommended because it can cause artifacts and is innacurate) if done probing side of spec is best
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| how is the bubble technique used to test DC endpoint | acid (with calcuim) will cause bubble formation, jar is shaken to remove, when no bubbles are present DC is done (UNRELIABLE)
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| How can weight loss/gain be used to check dc endpoint | weigh bone, DC, weigh (weight loss will = Ca loss, DC in fresh soln', repeat until their is weight gain indicating that there is water uptake in the bone
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| How is surface DC done during microtomy | (when DC not complete or soft tissue has Ca deposit) 1. rough cut block 2. put block face down in DC for 15-60 min., wash, resection
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