Lab
Quiz yourself by thinking what should be in
each of the black spaces below before clicking
on it to display the answer.
Help!
|
|
||||
|---|---|---|---|---|---|
| Cytopreparation | The act of preparing & staining diagnostic slides
🗑
|
||||
| Diagnostic cytology | Examination of cellular material for disease, esp. cancer
🗑
|
||||
| Gynecological cytology specimens | Pap smears, endocervical brushings,
collected & smeared on slide,
fixed while still wet, then dried & sent for staining
🗑
|
||||
| Non-gynecological cytology specimens | Urine, sputum, CSF, bronchial/gastric/esophageal washings or brushings, pleural/peritoneal/ascites/pericardial fluids, FNAs,
best collect fresh w/ no fixative/additive,
most can be refrigerated 24-72 hrs,
don't freeze
🗑
|
||||
| Pap smears | Liquid-based cytology rather than direct smear - better for automated systems - ThinPrep system & SurePath system,
smeared on slide, fixed while wet, dry & stain
🗑
|
||||
| ThinPrep System | Specimens w/o brush head in vial w/ weak methanol preservative,
homogenize & filter,
filter pressed on slide to deposit cells,
slide can be stained traditionally,
nongynecological specimens need extra suspension & centrifugation
🗑
|
||||
| SurePath System | Specimens w/ brush head in vial w/ very weak ethanol preservative,
cell enrichment - centrifuge conc. abnormal cells, remove blood/mucus,
specimen aliquot sediments on slides,
nongynecological req. alt. suspension & centrifugation,
higher yield cells
🗑
|
||||
| Preferred cytology fixative: | 95% ethanol,
rapid fixation important to prevent distortion of cell & loss of morphology,
🗑
|
||||
| Alcohol vs. formalin fixation | Alcohol better b/c crisp fixation of nuclear chromatin,
avoid formalin - causes different nuclear chromatin patterns which may impact diagnosis
🗑
|
||||
| Other cytology fixatives: | Commerical cytology sprays - usu. ethanol w/ acetone & polyethylene glycol (remove latter w/ alcohol before staining),
95% denatured alcohol,
80% isopropanol (b/c more shrinkage than ethanol),
100% methanol (b/c less shrinkage than ethanol)
🗑
|
||||
| Sparsely cellular specimens | Sample w/ not many cells in suspension can be made more conc'd by Cytospin or centrifugation preparation:
1mL properly prepared cell suspension in funnel of cytospin,
centrifuge,
cells will be deposited on slide & fluid absorbed by filter paper
🗑
|
||||
| Problem: bloody specimens: | Can obscure morphology or outnumber diagnostic cells,
crosshatch can help separate cells,
dip in Carnoy's or Clark to lyse rbc,
commercial products can lyse rbc (maybe diagnostic too)
🗑
|
||||
| Problem: poorly adhesive specimens | Often urine & breast fluid,
formalin-fixed may wash,
coated, positive-charged slides,
specimens prepared by centrifugation less likely to shed cells,
spray fixing (rather than liquid fixing) decr. slide cell loss
🗑
|
||||
| Problem: cross contamination | Ongoing concern, cells may float into reagents or stick to tools,
change reagents between samples & use disposable tools
🗑
|
||||
| Cell block | Can process, section, stain, as histo section,
easier special stain, incl. IHC,
made of material left after smear,
make from tissue fragments, clots, heavy mucus,
"block" together w/ thrombin-prothrombin clot, albumin, or agar
🗑
|
||||
| Fixation time: | Smears fixed w/i 1-2 seconds of application to slide,
thick smears/smears w/ mucus may take 2-4 seconds,
liquid suspensions longer,
air-drying causes nuclear swelling, distortion, loss of cytoplasmic density, cytoplasm becomes eosinophilic
🗑
|
||||
| EA stains metabolically inactive cells? | Various shades of pink
🗑
|
||||
| EA stains metabolically active cells? | Various shades of blue-green
🗑
|
||||
| Nickel smear method | Smear material from scraper/brush in circular motion onto slide in area the size of a nickel,
fix immediately w/ 95% ethanol or spray fixative,
avoid cotton swabs,
avoid feathered-edge smears
🗑
|
||||
| Pull-apart smear method | 2-3 drops of sediment on slide, place another slide on top upside-down, pull slides apart endways,
fix immediately w/ 95% ethanol or spray fixative,
good general-purpose for fluids w/o mucus
🗑
|
||||
| Crush/mash smear method | Mash material between two slides to give thin, even smear,
may use back & forth "slap" motion to break chunks,
fix w/ 95% ethanol or spray-fixative
🗑
|
||||
| Cross-hatch smear method | 2-3 loopfuls of sediment on slide, spread diagonally, back & forth, corner to corner,
fix immediately w/ 95% ethanol or spray fixative,
if sediment beads form, spread again for monolayer of cells,
good for cellular body fluids, esp. w/ lots of blood
🗑
|
||||
| Cytocentrifuge smear method | Specimen placed into funnel apparatus,
centrifugation forces specimen thru tunnel,
cells onto slide, filter absorbs fluid
🗑
|
||||
| To evaluate cellularity of specimen to determine adequacy: | Toluidine blue wet film,
or air-dry & stain w/ Diff-Quik
🗑
|
||||
| Problem: Pap-counterstained smears look dirty & not optimal: | Ensure clean alcohol rinses after counterstains,
overused alcohol rinses may cause "dirty" colors
🗑
|
||||
| Bodys fluids pleural, peritoneal, pericardial, ascites treatment: | Send entire amount collected to lab,
good for cell block
🗑
|
||||
| Breast/nipple discharges treatment: | Prepare smear at time of collection,
nickel smear,
spray-fix, don't let air-dry
🗑
|
||||
| CSF treatment: | Deteriorates rapidly,
if can't take to lab immediately use pre-fixative holding solution - alcoholic saline or Saccomanno fluid = vol. of specimen
🗑
|
||||
| Direct scrapings for viral lesions (Tzank smears) treatment: | Prepare smear at time of collection,
nickel smear,
spray-fix, don't let air-dry
🗑
|
||||
| Washings - bronchial, esophageal, & gastric treatment: | Don't fix, refrigerate immediately
🗑
|
||||
| Urine treatment: | V. fragile, doesn't hold pH,
first morning urine not recommended b/c cells degenerated,
add pre-fixative if can't process or send to lab right away,
equal volume of alcoholic saline or Sacomanno fluid
🗑
|
||||
| Brusings - bronchial, esophageal, & gastric treatment: | Prepare smear at time of collection,
nickel smear,
spray-fix, don't let air-dry,
or submit brush in physiological saline for lab processing,
never fix
🗑
|
||||
| FNAs include: | Breast, thyroid, lung, liver, pancreas,
cell-laden fluid removed from site,
initially use pull-apart method & fix,
never feathered-edge,
rinse needle w/ physiologic saline (don't rinse w/ fixative) to recover remaining specimen
🗑
|
||||
| Alcoholic saline pre-fixative reagents | 1 part saline
1 part 50% alcohol
🗑
|
||||
| Saccomanno Fluid pre-fixative reagents | 980 mL ethanol
20 mL melted Carbowax
(remove Carbowax after slide prep, before staining)
🗑
|
||||
| Cytology smears: | Fresh, unfixed specimens, wet-fixed on slide,
better cell adhesion,
causes cells to flatten - better visualization,
pre-fixed specimens - round cells
🗑
|
||||
| Direct smears include: | Scrapings, brushings, viral inclusion smears,
use nickel method
🗑
|
||||
| Fluids include: | Pleural, peritoneal, pericardial, ascites, effusions, urine, cyst fluids, washings,
first centrifuge to concentrate cells,
use pull-apart, or cross-hatch method
🗑
|
||||
| Best centrifugation for cytology: | 2,000 rpm for ~10 min,
high speed packs cells too tightly,
lower speed won't force cells to bottom of tube,
50 mL conical tube best (15 mL usu. not adequate specimen),
swinging-arm centrifuge best concentrates cells @ bottom
🗑
|
||||
| Mucoid specimens include: | Sputum & thick bronchial washings,
use crush method
🗑
|
||||
| Sparsely cellular specimens include: | CSF, sparsely cellular urines or body fluids,
filter w/ cellulose or polycarbonate filter (5 µm pore) to capture all cells, stain & mount filter (old method),
more common - cytocentrifugation
🗑
|
||||
| Cytopreparation for sample with mucus? | Crush method w/ spray fixative
🗑
|
||||
| Cytopreparation for sample with good cellularity & adhesiveness? | Pull apart or crosshatch w/ spray fixative or 95% alcohol
🗑
|
||||
| Cytopreparation for sample with good cellularity & poor adhesivenss? | Pos slide or cytocentrifuge w/ spray fixative
🗑
|
||||
| Cytopreparation for sample with poor cellularity? | Cytocentrifuge
🗑
|
||||
| Cell block for specimen w/ spontaneous clots: | Squeeze out excess liquid until solid mass,
place mass in labeled cassette, may wrap w/ lens paper,
if too large submit portion,
formalin process routinely
🗑
|
||||
| Cell block for specimen w/ cellular material entrapped in mucus: | To remaining sediment add 95% ethanol, denatured alcohol, or 10% NBF, mix by inversion,
fix 15-30 min,
centrifuge 10 min, 2,000 rpm,
pour off supernatant,
mass in cassette, may wrap w/ lens paper,
if large submit portion,
formalin process routinely
🗑
|
||||
| Cell block for specimen w/ loose cellular material - albumin method: | To remaining sediment add 30% BSA, mix w/ stick,
add 95% alcohol,
place mass in cassette, may wrap w/ lens paper, if too large submit portion,
formalin process routinely
🗑
|
||||
| Cell block for specimen w/ loose cellular material - agar method: | To remaining sediment add melted 4% agar, mix gently,
centrifuge 10 min, 2,000 rpm,
refrigerate until agar solid,
remove from tube, cut away cellular material, bisect,
place in cassette w/ common cut side down,
formalin process routinely
🗑
|
||||
| Papanicolaou technique stains: | Hematoxylin nuclear stain, Harris or Gill,
OG-6 counterstains tonofilaments in keratinized cells,
EA differentially counterstains cytoplasm of cells,
store all in light-blocking containers
🗑
|
||||
| OG-6 reagents | Orange G
dH2O
phosphotungstic acid (mordant)
95%ethanol
glacial acetic acid (opt.) (enhance specificity, decr. stain time)
🗑
|
||||
| EA reagents | eosin Y 20%
light green SF 3%
phosphotungstic acid
95% ethanol
abs. methanol
glacial acetic acid
Bismarck brown (may be excl'd makes stain weaker)
🗑
|
||||
| Pap stain purpose | To distinguish cellular components by obtaining highly detailed chromatin, differential counterstaining, & cytoplasmic transparency
🗑
|
||||
| Pap stain reagents | hematoxylin
ammonia water, or Scott tap water
OG-6
EA
🗑
|
||||
| Pap stain results | chromatin - blue
keratin - orange
superficial squamous cells, nucleoli, cilia, rbc - variable pinks
all metabolic cell cytoplasm - variable blue-greens
🗑
|
||||
| Toluidine blue wet film purpose | Rapid determination of cellularity, esp. FNAs,
& eval. potential cross-contamination
🗑
|
||||
| Toluidine blue wet film facts | Gives metachromatic staining in unfixed cells,
use unfixed cytological specimen
🗑
|
||||
| Toluidine blue wet film reagents | Toluidine blue in 95% alcohol w/ dH2O, filtered
🗑
|
||||
| Cross-contamination in cytology specimens: | Significant chance of loose cells floating during staining,
to prevent stain separately specimens w/ higher potential to float, filter solutions between batches
🗑
|
||||
| What is a LEEP biopsy? | Loop electrosurgical excision procedure,
cervix biopsy
🗑
|
Review the information in the table. When you are ready to quiz yourself you can hide individual columns or the entire table. Then you can click on the empty cells to reveal the answer. Try to recall what will be displayed before clicking the empty cell.
To hide a column, click on the column name.
To hide the entire table, click on the "Hide All" button.
You may also shuffle the rows of the table by clicking on the "Shuffle" button.
Or sort by any of the columns using the down arrow next to any column heading.
If you know all the data on any row, you can temporarily remove it by tapping the trash can to the right of the row.
To hide a column, click on the column name.
To hide the entire table, click on the "Hide All" button.
You may also shuffle the rows of the table by clicking on the "Shuffle" button.
Or sort by any of the columns using the down arrow next to any column heading.
If you know all the data on any row, you can temporarily remove it by tapping the trash can to the right of the row.
Embed Code - If you would like this activity on your web page, copy the script below and paste it into your web page.
Normal Size Small Size show me how
Normal Size Small Size show me how
Created by:
CCF
Popular Histology sets