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UND 363 Cyto/QC

UND 363 Cytology and QC

cytology is examination of cellular material
gynecology cytology is study of cellular material from female genital tract
non-gyn cytology study of cellular material from various body sites (sputum, urine, csf)
a cell block is concentration of cells that can be used in standard histology
cytocentrifugation is spinning of cellular material to seperate fluid and cells (used for scantly cellular specs (urine/csf)
bluing is changing ph above 8.0 so the stain will change from red to blue
cytology vs histology cytology looks at individual cells and morphology while histology looks at tissue architecture
adv vs. disdv of pap stain adv-good cellular detail of nuclear and cyto morphology. DIS - not standardized, not specific for any compound for malignancy and has a loss of cells
thin prep vs. surepath collection methods both collect specs on broom or brush, but surepath requires the brush sent in the vial whereas thinprep does not
name three types of fixation spray fixing, wet, and airdrying
what fixatives are used in cytology 95% alcohol, 80% isopropanol and 100% methanol can be substituted for 95% alcohol, spray fixatives
what do the fixatives do in fixation keeps cells in place for staining. preserves and hardens the cells, and removes/replaces water, stops autolysis and sterilizes
how are spray fixatives different than the "wet" fixatives contain alcohol and wax (the wax must be removed prior to staining by using 95% alcohol or poor stains may result). alcohol fixes and the wax forms protective coating
how can carnoy and clark fixatives be used and for what type of specimin used for bloody specimens. They lyse RBC's so that the diagnostic cells can be more visible. a drawback is that they damage the cell membrane
what does the saponin procedure help fix bloody specimens - it adds hanks or isotonic solution & saponin. it is then centrifuges. This removes hemoglobin and leaves nucleated cell pellet.
how is the sureprep method done weak ethyl alcohol solution. centrifuged and the sediment is put onto a slide. surepath also stains each slide individually. requires the brush be sent therefore no swishing.
how is the thinprep method done weal methanol solution. uses a filtration method where negative pressure draws up the cells on to a filter and the cells are stamped on a slide. does not want the brush.
what are the sizes of specimen areas for surepath vs thinprep surepath is 13mm, thinprep is 20mm
what cyto technique uses a blender and for what specimens saccamanno for sputum
What stain in cytology is polychromatic EA
what are the two clearing agents used in cytology xylene and clearium (clearium allows slide to be taken from last alcohol to coverslipping - combines clearing and mounting)
What stain can be used to determine if malignant cells are present prior to the Pap stain? toluidine blue
what are the techniques for fluid processing centrifuge, direct smears (scrapings, brushings tzank smears), stripe technique, pull apart, cell block, nickel method, crush method (similar to pull apart but has back and forth motion to get thin layer - for mucus)
what stains are used in the pap stain harris or gill hematox, OG6, EA (combining eosiny, lt green SF or fast green, and bismark brown)
what is the staining order of non gyn specimens (first to last) start with most scantly cellular fluids (csf then urines), effusions should be last, and FNA should be near end as well since they are usuall malignant.
why should csf and urine be stained quickly CSF cells deteriorate rapidly and the ph of urine will destroy cell morpholygy
what is the difference between QC and QA QC - defines quality, is prospective, and deals with all processes and outcomes. QA - measures success is RETROspective, uses sample outcomes and impacts processes
what are the colors associated with basophilic, orangiophilic, indeterminate, acidophilic, cyantophilic blue, orange, gray blue, pink, blue-green
what are 5 other clia regulation for cytology specs besides the slide and report requirements labelling of source/confirmation of specimin, specs from and through authorized personnekl, processing request (which test to be done), date of obtainment, patient info
what are some methods of direct smears scraping, brushing, tzank smear, viral inclusions, nickel method
what is the storage time for gyn/non gyne specs, FNA's, and the records and reports 5 years for gyn/non gyn, 10 yrs for FNA, records and reports (10 yrs)
what is the difference between gyn and non gyn cytology nongyn is a diagnostic process whereas gyn is a screening
what happens to cell shape when wet fixed (in liquid)vs. fixed onto a slide/air dried wet fixed (in liquid) the cells will be rounded, if fixed to a slide they will flatten out
what are the variables when processing liquid specimens fresh or not, alcohol preserv used? (equal parts fix to spec), volume/cellularity, spec type, additional processing needed (is it bloody, mucousy)
if a spec has low cellularity and low volume what should be used cytocentrifugation cytospins (special centrifuge that deposits cells to be direct deposited on slide with residual fluid absorbed on filter paper.
if a spec has low cellularity and high volume what tech should be used centrifugation with removal of supernatant and then resuspension of the button and cytospins (ex. urine)
if a spec has high cellularity and high volume what tech can be used centrifugation, resuspension of button and smear prep (ie washings)
if a spec has high cellularity and low volume what tech should b used centrifugation and resuspension of the button, smear, Surepath and thinprep may be used
explain centrifugation spin and pour off supernantant, resuspend button and pipette out for slide
what does a vortex mixer do resuspends the cell button for homogeneity since centrifugation can layer the cells depending on their mass
what are some techniques for smears nickel method - smear with brush etc. from inside out in circle to make a nickel size spec. pull apart - slide over slide facing each other with spec inbetween and pull apart. crosshatch - use loop and spread back and forth down slide (zigzag)
what are the methods to make cell blocks fixed sediment, agar cell block, tea bag tech
what is the tea bag cell block method centrifuged button placed in tissue bag, formalin and processed normally for histology
what is the strip technique for effusions centrifuge, lyse blood if needed, decant supernantant, remove bufferlayer with cotton tip (NO resuspension), press q-tip in one line down slide and fix in alcohol for 30 min.
what are the two methods for mucoid specs pick N smear - white and bloody flecks places on slied and use pull apart/crush method. Sacamanno - preserved in carbowax and ethanol and homogenized in blender then centrifuged, etc..
why must effusions (body cavity fluids) be stained seperately the cells have high floating probablility and may not stick to slide
why must CSF be centrifuged with slower acceleration cells are delicate
how is the toluidine blue staining method done for IDing malignate cells so they are processed seperately one drop of sample on slide, put second drop of stain next to sample and mix, coverslip and bring to pathologist.
what are some drawbacks to pap stain unpredictable stain (auto stainers have mostly removed this) and loss of cells during processing
what does EA stain cytoplasm, nucleoli, and cilia
what fixation cannot be used for pap stain air drying - causes nuclear swelling and loss of cytoplasmic and nuclear detail
what stain can be used if a fluid has been airdried diff quik (ie FNA)
what are the two nuclear stains for the pap stain and how are they used gills - sodium iodate oxidizer and aluminum mordant - progressive PH 2.34, Harris, regressive stain and more concentrated
what are the bluing agents in pap staining tap water, ammonium hydroxide, lithium carbonate, scotts tap water substitute.
give details of orange G6 (OG6) in pap stain first coutnerstain, acidic, it is "trapped" in dense cytoplasm (doesn't combine), monchromatic, colors keratin brilliant orange
what is a drawback to OG6 and what needs to be done must rinse well or OG6 will get trapped in other areas wich will obscure stain
What is the second counterstain in Pap stain EA (Eosin Azure), polychrome stain, EOSIN Y stains cytoplasm or mature squamous cells, nucleoli, RBC's and cilia, LIGHT GREEN - stains cells that are metabolically ACTIVE, bismark brown left out of newer formulas
what are the EA formulations that are used in gyn and nongyn staining EA50 for Gyn, and EA 65 for nongyn
what is the ph of EA 4.5 - 5.5
how is the pap stain different from other staining fixed cells use one step hydration and dehydration (direct from 95 to water). fixed cells don't require graduation.
what will happen to lt green if the spec is not in EA long enough the lt green will not penetrate since it is in a smaller concentration than eosin
what will happen if the spec is in OG too long it can mask the eosin. BUT the lt green will still show,.
what is cell enrichment combines gravity dispersion and centrifugation to seperate obscuring problems (blood, mucus, inflammation) from diagnostic material
what are 4 steps in surepath collection (with brush in vial), cell enrichment, cell on slide, automated staining
what are some pros and cons of surepath 13mm cell layer, reagents never shared, stain incorporation CONS - lack of stain control, not fda approved, smaller cell area, slides must be coated correctly or machine will not work well.
thinprep steps collection with broom (broom rinsed in container and discarded), sent to lab, gentle dispersion and mixing, negative pressure draws fluid through filter and then filter stamped on slide, fixed. stained SEPERATE from machine
pros/cons of thin prep pap staining pro- uses lab staining, 20mm spec area, HPV TESTING (surepath does NOT), con - methanol preservative, hyperchromasia is less pronounced (less color)
thinprep body fluid process cytolyt collection, centrifuge, removal of supernantant, add cytolyte wash, spec in preservcyt solution, run on machine
how is mucoid done in thinprep adds mechanical agitation.
how do the machines differ in regards to automation of points of interest thinprep stores coordinates of interest points, the surepath does a ranking system whether the slide should be looked at at all.
what are the albumin method and agar method used for to be added to loose cellular material so that they can be put in a cassette and processed in formalin. albumin forms mass and then put in lens paper, agar solidifies and is cut in half and put in casette.
what can be added to OG6 for enhancment of specificity and decreased stain time acetic acid
Created by: mustangvxd



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