Morphology I Slide: 1 - Cytology stuffcytologystuff.com/pdfs/ADS-00690 TPPT Morphology Lecture...
Transcript of Morphology I Slide: 1 - Cytology stuffcytologystuff.com/pdfs/ADS-00690 TPPT Morphology Lecture...
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Morphology I Slide: 1
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Morphology I Slide: 2
ThinPrep Morphology
Normal Cytology
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Morphology I Slide: 3
CT & Pathologist Training
Training program begins with ThinPrep morphology presentation
• Microscopic training sessions for individual and multi-headed group
screening sessions are designed to develop and refine screening and
interpretive skills.
• Screening Evaluation modules provide more thought-provoking
cases intended to provide a full range of diagnostic challenges.
• Competency Assessment modules are used to monitor training
participants’ performance (pre- and post-tests).
• Laboratory training is the final phase of training within the
laboratory. The goal for the Pathologist/CT is to transfer conventional
pap knowledge and confidence by screening and diagnosing ThinPrep
slides. Ongoing laboratory training material (glass slides) comes from
ThinPrep Pap Test collection in the laboratory.
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Morphology I Slide: 4
ThinPrep Process
Three key phases
1. Dispersion: Randomizes/homogenizes patient’s cell population
within the vial.
2. Cell Collection: ThinPrep software monitors the flow rate and
senses when pores in the filter are blocked by material (red blood cells
[RBCs], white blood cells [WBCs], and epithelial cells).
3. Cell Transfer: The cylinder is inverted and the filter comes in
contact with the glass slide. Air pressure from behind the filter aids the
cells’ natural attraction to the glass slide.
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Morphology I Slide: 5
Conventional Pap Smear (Macroscopic)
• The limitations of the manual smearing method are apparent, with
variable thickness of the smear. Fortunately, conventional slide
morphology is transferable to the ThinPrep process. Cytology
knowledge does not have to be relearned, base knowledge is merely
refined.
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Morphology I Slide: 6
ThinPrep Pap Test (Macroscopic)
• A key difference in presentation is the absence of a manual smear
pattern. The ThinPrep process takes the patient sample and applies it
onto the center of the slide in a thin, uniform layer. Specimen
preparation is standardized, eliminating the inconsistency associated
with manual preparations.
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Morphology I Slide: 7
Conventional Pap (CP) smear
• Microscopically, the uneven distribution of cellular material
associated with the CP smear pattern is evident.
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Morphology I Slide: 8
ThinPrep (TP) Pap Test
Same Patient
• Tissue architecture is maintained but the ThinPrep process rearranges
the relationship of cell groups and provides a more even distribution of
cells and groups on the glass slide. Note the group/sheet of intact
endocervical cells.
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Morphology I Slide: 9
TP Characteristics
Common changes associated with TP morphology
• Liquid Based Fixation - The key difference.
• Cell Size - Related to fixation and the effect of cells placed into
solution.
• Smear Pattern - No longer smearing cellular material across the
glass slide.
• Specimen Background - Unique characteristics of rinsing cells into
a solution but “clues” are still present.
Most important, the similarities between the TP and CP far outweigh
the differences.
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Morphology I Slide: 10
TP Characteristics
Liquid Based Fixation
• Enhanced cytoplasmic detail – Optimized fixation, lack of
mechanical (smearing) artifact.
• Enhanced nuclear detail - Optimized fixation, lack of mechanical
(smearing) artifact.
• Variability in nuclear staining – The chromatin detail is more readily
appreciated and can appear as hypo- and hyperchromatic nuclei within
the same case.
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Morphology I Slide: 11
TP Characteristics
Cytoplasmic Detail
• Koilocytosis caused by HPV is often more evident than in
conventional paps and the spectrum of eosinophilic and cyanophilic
staining is seen. Mechanical artifacts are absent.
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Morphology I Slide: 12
TP Characteristics
Nuclear Detail
• Dark, hyperchromatic nuclei are still identified on TP, as illustrated
in this HSIL cell.
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Morphology I Slide: 13
TP Characteristics
Variability in Nuclear Staining - ThinPrep Stain
• Left image: Dependent upon patient biology, the chromatin of the
dysplastic cells can appear hypochromatic. However, increased N/C
ratios and irregular nuclear membranes should alert you to an abnormal
process, as in this case of HSIL.
• Right image: These HSIL nuclei show the hyperchromatic chromatin
pattern typically associated with dysplasia.
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Morphology I Slide: 14
TP Characteristics
Variability in Nuclear Staining – Richard-Allan stain
• Left image: Here is another example of hypochromatic appearing
nuclei in a case of HSIL stained with Richard-Allan stain. It is
important that these cells are not missed, regardless of the stain used.
• Right image: These HSIL nuclei show hyperchromatic chromatin.
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Morphology I Slide: 15
TP Characteristics
Cell Size
• “Proportionally” smaller cells - Due to the inherent difference with
alcohol fixation and the elimination of air-drying artifact cells may
appear smaller but will be in proportion to the other cells on the slide.
• Single cells are more prominent - Single cell populations are not
created, just more noticeable in the background.
• Cells round up in solution – Due to the physical properties related to
suspension in fluid, the changes are similar to a non-gyn presentation.
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Morphology I Slide: 16
TP Characteristics
Single Cells
• Single cells and small groups are easily seen because of the thin layer
presentation. These include endocervicals, endometrials and squamous
metaplastic cells.
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Morphology I Slide: 17
TP Characteristics
Proportionately Smaller/Rounded Up
• The squamous metaplastic cells in this cluster may appear to be
smaller than those on conventional paps but are proportional in size to
the mature squamous cells in the background. There is also more depth
of focus to the group due to the rounding up which occurs in solution.
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Morphology I Slide: 18
TP Characteristics
Smear Pattern
• Mechanical artifacts associated with pulling the cells across the slide
have been eliminated.
• Cellular material is not pulled out in mucus, therefore there is a new
pattern recognition that needs to be established for TPs. Endocervical
cells in particular will be distributed throughout the cell deposit and are
not caught up in mucus.
• Tissue architecture is maintained throughout the slide processing.
Architecture is not disrupted, only the relationships of the cell
groupings have been altered.
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Morphology I Slide: 19
TP Characteristics
CP – Smear Pattern
• This slide exhibits a common presentation of cells pulled or streaked
across the slide with areas of obscuring inflammation and mucus.
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Morphology I Slide: 20
TP Characteristics
TP – Same Patient
• Notice that the cells are evenly distributed throughout the
background, not pulled out in mucus.
• Mechanical artifact is eliminated.
• White cells, indicating that inflammation is still present, can be seen
in the background.
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Morphology I Slide: 21
TP Characteristics
Specimen Background
• Specimen background appears cleaner as a result of the ThinPrep
process.
• Fluid collection may cause cellular debris to clump.
• Background can provide clues. TP requires more vigilance when
screening because the cellular and non-cellular elements typically
obscured on the CP are more easily identified on a TP slide and there is
more to see per field.
• “Ratty” background can be advantageous during screening and
infectious agents, cytolysis and/or disease should be considered. Tumor
diathesis, blood and cytolysis will still be present on the slide and
should be used in the differential diagnosis.
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Morphology I Slide: 22
Specimen Background
Cytolysis
• Bare nuclei and cytoplasmic debris are evenly dispersed across the
cellular deposit. Döderlein bacilli typically seen on top of squamous
cells or trapped in mucus are still present.
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Morphology I Slide: 23
Specimen Background
Blood
• RBCs appear as ghost-like cells as a result of the blood lysing
properties of the preservative solution. Hemolyzed blood is often found
clumped.
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Morphology I Slide: 24
Specimen Background
Trichomonas vaginalis
• Well preserved organisms along with a bacterial background can give
the appearance of a “ratty” specimen.
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Morphology I Slide: 25
Specimen Background
Tumor Diathesis
• Tumor diathesis retains the characteristic appearance of cell debris,
protein and blood in the background in addition to WBC’s. It also
maintains the dirty/gritty background appearance seen on a CP.
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Morphology I Slide: 26
Microscopic Evaluation
When Screening Slides:
• Systematic approach - Screening vigilance is required over a smaller
area with an increased attention to the background where potential
single abnormal cells may be found.
• Slow approach – A natural tendency may be to screen fast because of
the cleaner appearance of a TP but a slow, steady screening technique
allows time to incorporate subtle background clues. Cleaner
background does not necessarily indicate normal cell population.
• Slight overlap – This technique is still required to ensure that as
many cells as possible can be visualized during screening.
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Morphology I Slide: 27
Specimen Adequacy
Cellular Composition
Specimen adequacy guidelines for Liquid Based Cytology as
recommended by the Bethesda System 2001 are:
• 5000 well-visualized and preserved squamous cells are necessary for
a satisfactory specimen.
• Strict objective criteria may not apply in every case. Slides with cell
clustering, atrophy or cytolysis are technically difficult to count and
laboratories should apply professional judgment and employ hierarchical
review when evaluating these slides.
• The presence or absence of an endocervical component should be
noted. Ten well-preserved endocervical or squamous metaplastic cells, singly
or in groups, should be observed to report a transformation zone component as
present.
• The presence of abnormal cells makes any specimen an adequate
specimen.
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Morphology I Slide: 28
Depicted is a 10X field containing 60 squamous cells.
• Satisfactory field @ FN22
• TBS requirements are:
- 10 fields across cell deposit diameter should have an average of 60
cells each.
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Morphology I Slide: 29
Depicted is a 10X field containing 50 squamous cells.
• Satisfactory field @ FN20
• TBS requirements are:
- 10 fields across the cell deposit diameter should have an average of
50 cells each.
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Morphology I Slide: 30
Depicted is a 10X field containing 40 squamous cells.
• Unsatisfactory field due to insufficient squamous epithelial
component
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Morphology I Slide: 31
Cellular Composition
Unsat/Blood
• Predominantly blood was rinsed into this PreservCyt solution vial.
Microscopically only lysed blood, rare epithelial cells, and ghost RBCs
are identified and this slide would be unsatisfactory for interpretation.
Communication between the lab and the clinician remains key in
troubleshooting unsat specimens.
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Morphology I Slide: 32
Cellular Composition
Unsat/WBCs
• RBCs, WBCs and epithelial cells all compete for a place on the filter,
so if there is a heavy inflammatory population, that will alter the
composition of the final TP slide. The slide is still representative of
what the clinician has collected and rinsed into the vial.
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Morphology I Slide: 33
Cellular Composition
Unsat/Mucoid
• Mucus will compete with cells for a spot on the filter. A truly mucoid
specimen will show islands or pools of mucus and may affect the
specimen adequacy.
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Morphology I Slide: 34
Endocervical Cells
• Honeycomb/palisading formations are maintained.
• Cells tend to round up in solution.
• More tightly packed cell groupings with cell groups twisted or folded
from the fluid collection method may be seen.
• Smaller endocervical cell groupings as well as single cells may be
present and careful screening is required to identify them.
• Liquid based fixation results in nuclei that appear busier, more
reactive.
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Morphology I Slide: 35
TP Characteristics
Endocervicals
• Large sheet of endocervical cells exhibiting both honeycomb and
picket- fence arrangements.
• Nuclei are small and uniform.
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Morphology I Slide: 36
TP Characteristics
Endocervicals
• These cells exhibit traditional palisading formation with small,
basally located nuclei.
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Morphology I Slide: 37
TP Characteristics
Endocervicals
• Small, cuboidal group of endocervical cells with round to oval
uniform nuclei can be seen. Infrequent, small cell groupings may be
more challenging to find and identify.
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Morphology I Slide: 38
Squamous Metaplasia
• Traditional sheets and cobblestone arrangements are presented.
• Typical dense, homogenous cytoplasm is evident. Increased
cytoplasmic vacuolization may be present as a result of the liquid based
fixation.
• Metaplastic cells can often occur singly.
• Cells appear smaller and rounder as a result of the fluid collection.
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Morphology I Slide: 39
TP Characteristics
Squamous Metaplasia
• Small grouping of squamous metaplastic cells with dense cytoplasm,
cytoplasmic vacuolization, and uniform, round nuclei.
• “Pseudopodia” or cytoplasmic processes identified on the
conventional smear are still evident even with the tendency of cells to
round up in solution.
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Morphology I Slide: 40
TP Characteristics
Squamous Metaplasia
• As a result of the liquid fixation, squamous metaplasia may present
as larger sheets with more depth to the groupings. There are prominent
cytoplasmic borders with some cytoplasmic vacuolization and round to
oval nuclei with smooth nuclear membranes, fine, even chromatin and
micronucleoli.
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Morphology I Slide: 41
Endometrial Cells
• Tight 3D cell clusters are maintained.
• More loose cell groupings with cytoplasmic vacuolization may be
identified.
• Single cells are more prominent against a cleaner background.
• Nuclei appear busier as a result of the improved preservation in the
liquid based fixative.
• Menstrual blood in the background is lysed, and may appear
clumped.
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Morphology I Slide: 42
TP Characteristics
Endometrials
• Menstrual background - RBCs appear hidden in background as ghost
blood cells as a result of the lysing properties of the preservative
solution. This lysed blood may have a tendency to present in clumps.
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Morphology I Slide: 43
TP Characteristics
Endometrial Cells
• Tight cell group with endometrial and stromal cells typical of an
exodus pattern.
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Morphology I Slide: 44
TP Characteristics
Endometrial Stromal Component
• A small loose cluster with kidney bean shaped nuclei and vacuolated
cytoplasm. Note the size relationship of the stromal cells to the
intermediate cell nuclei.
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Morphology I Slide: 45
Look-Alike
Endocervical vs. Endometrial
• Left image: The grouping of small endocervical cells presents in a flat
sheet with nuclei of equal size and shape.
• Right image: The endometrial cells maintain a 3D configuration and
the nuclei may vary in shape but do not vary in size.
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Morphology I Slide: 46
Atrophy
Preservation is greatly enhanced as a result of the liquid based fixation
with the elimination of the air-drying artifact commonly seen on CP.
• Consistently observe sheets of well preserved parabasal cells.
• It is possible to distinguish endocervical cells from parabasal cells,
often something that was difficult on a conventional slide.
• The number of bare nuclei are reduced by removing the mechanical
artifact of smearing.
• “Atrophic vaginitis” background pattern appears more clumped as a
result of the fluid collection method.
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Morphology I Slide: 47
Parabasal Cells
• Parabasal cells may appear in uniform large sheets, with easily
identifiable cytoplasmic borders, abundant cytoplasm and round to oval
nuclei. Sheets may be twisted or folded over during the preparation
process.
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Morphology I Slide: 48
Atrophy
• Sheets and single parabasal cells are readily identified.
• Stripped parabasal nuclei are reduced but not eliminated.
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Morphology I Slide: 49
Atrophy
• Squamous component may be easily distinguished from the
honeycombed endocervical component.
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Morphology I Slide: 50
Atrophic Vaginitis
• The background pattern presents as clumped cellular debris and
degenerated WBCs encircling the parabasal cells.
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Morphology I Slide: 51
Trichomonas vaginalis
• Organisms may appear smaller as a result of the liquid based fixation.
• It is easier to identify the internal structure to differentiate
cytoplasmic debris from an organism.
• The classic “Trich” pattern is maintained and inflammatory cell
changes are still identifiable.
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Morphology I Slide: 52
Trichomonas vaginalis
• Inflammatory cell changes, including peri-nuclear halos, cytoplasmic
vacuolization and bichromatic staining are useful in pattern recognition.
The well preserved organism can readily be distinguished from
cytoplasmic debris (look-alike) by visualizing the elongated eye-spot
within the pear-shaped organism. Flagella may also be preserved.
(Compare the trichomonad to the adjacent white cell).
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Morphology I Slide: 53
Trichomonas vaginalis
(Screening Power)
• Even in the absence of inflammatory cellular changes, the organisms
(about the size of the WBCs) can be noted in the background.
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Morphology I Slide: 54
Candida Spp.
• Classic cell clumping is more evident.
• Reactive squamous cells with engulfed WBCs may be present as a
clue.
• It is important to differentiate between mucus strands and
pseudohyphae.
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Morphology I Slide: 55
Candida Spp.
(Low Power)
• Organism can be seen isolated in the clean background or in the
traditional “shish-kabob” or “spearing” arrangement.
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Morphology I Slide: 56
Reactive Changes Due to Inflammation
• Reactive squamous cells with engulfed WBCs associated with
Candida spp. often serve as a clue and present with vacuolated “tissue
paper” or “moth-eaten” cytoplasm and reactive nuclei.
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Morphology I Slide: 57
Look-Alike
Candida vs. Mucus
• Left image: Candida, like mucus strands, can be twisted within the
cell groupings. However, the pseudohyphae can be identified
projecting from the groupings and distinct cell walls can be seen.
• Right image: Mucus strands can be seen extending from cell groups
but, upon close inspection, the strands are irregular in width and
distinct hyphae and spores are not present.
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Morphology I Slide: 58
Candida Spp.
• Fungal spores are retained and tend to clump together or pile on top
of cells.
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Morphology I Slide: 59
Actinomyces
• Organized clusters of branching filamentous bacteria.
• Associated blue staining bacteria are present.
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Morphology I Slide: 60
Actinomyces
• Classic blue staining bacteria associated with the branching
filamentous bacteria can be seen.
• Most often associated with patients with an IUD.
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Morphology I Slide: 61
Herpes Simplex Virus
Classic morphology, cells groupings are clearly noted from screening
power, isolated by the relatively clean background.
• Diagnostic ground glass nuclei are present with chromatin
margination and sharply defined nuclear membranes.
• Multinucleation with nuclear molding is easily appreciated.
• Eosinophilic nuclear inclusions, when present, are evident.
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Morphology I Slide: 62
Herpes Simplex Virus
• Multinucleation with nuclear molding and ground glass nuclei are
seen in these cells. Nuclear inclusions are more prominent with liquid
based fixation. Drying and/or mechanical artifacts are eliminated.
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Morphology I Slide: 63
Inflammation
It is important to use all criteria for a diagnosis.
• Slight nuclear enlargement (2x intermediate cell nuclei).
• Perinuclear halos – Important to distinguish from koilocytosis
associated with HPV changes.
• Cytoplasmic vacuolization and bi-chromasia may be present, similar
to morphology as seen on CP.
• Clumped WBCs in background are more visible.
• Enhanced cellular detail is present due to the improved fixation and
preparation.
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Morphology I Slide: 64
Inflammatory Changes
• These cells exhibit a slight increase in nuclear size and
hyperchromasia, with some nucleoli present. Perinuclear halos and
bi-chromasia are also noted. Nuclear changes fall short of those
required for an ASC-US diagnosis.
• Slight nuclear membrane irregularities are somewhat uniform in
appearance and are a direct result of cytoplasmic degenerative
changes.
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Morphology I Slide: 65
Repair
• Sheets of epithelial cells are present with engulfed WBCs.
• Cell groupings are more rounded and display a greater depth-of-focus
due to the liquid based fixation.
• Mechanical artifact is eliminated and the cells don’t appear as
“pulled out.”
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Morphology I Slide: 66
Repair
• Nuclei in this group of repair show bland chromatin, regular nuclear
membranes, nucleoli, uniformity within the sheet, and maintain nuclear
polarity.
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Morphology I Slide: 67
Look-Alike: Repair vs. Carcinoma
• Repair maintains a sheet-like configuration with a tissue-culture
appearance to the cytoplasm. Nuclei may be enlarged, however,
chromatin pattern is evenly distributed and nuclear borders are smooth.
Nuclei maintain a polarity to each other in these groups. Nucleoli are
present, may be prominent but are not in every nucleus.
• Carcinoma shows a greater depth of focus to the cell cluster and
greater variability from nucleus to nucleus with a marked alteration
in polarity. Nucleoli will be prominent and often multiple.
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Morphology I Slide: 68
ThinPrep Morphology
Epithelial Cell Abnormalities
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Morphology I Slide: 69
TP Characteristics
Common changes associated with TP morphology
• Liquid Based Fixation - The key difference.
• Cell Size - Related to fixation and the effect of cells placed into
solution.
• Smear Pattern - No longer smearing cellular material across the
glass slide.
• Specimen Background - Unique characteristics of rinsing cells into
a solution but “clues” are still present.
Most important, the similarities between the TP and CP far outweigh
the differences.
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Morphology I Slide: 70
ASC-US
ThinPrep reduces preparation artifact associated with some ASC-US
• Cells are found in sheets or singly
• Nuclei 21/2 – 3 times the size of an intermediate nucleus
• Uniform chromatin distribution
• Decrease in preparation artifact due to liquid based fixation
As a general guide, the frequency of ASC-US diagnoses should not exceed
2 – 3 times the rate of SIL.
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Morphology I Slide: 71
ASC-US
• The increase in nuclear size, increased hyperchromasia and
chromatin clumping shown here qualitatively and quantitatively falls
short of true LSIL.
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Morphology I Slide: 72
ASC-US
• Small group of squamous cells exhibiting slight nuclear enlargement
and a suggestion of HPV.
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Morphology I Slide: 73
ASC-US
• A group of squamous cells representing atypical parakeratosis with
slightly enlarged, irregular, hyperchromatic nuclei.
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Morphology I Slide: 74
ASC-H
• Immature metaplastic cells with increased N/C ratios, slight nuclear
membrane irregularities and nuclear size variability, suggestive of but
not definitive for HSIL.
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Morphology I Slide: 75
LSIL
With ThinPrep, the decision between normal and abnormal is more
straightforward. The ASC-US category still exists but the liquid based fixation
allows for better visibility of cellular criteria and may result in a definitive
LSIL diagnosis rather than an equivocal ASCUS diagnosis.
• Increased nuclear detail results in more chromatin detail visible
within nuclei.
• Nuclear membrane irregularities are more evident, otherwise lost or
obscured on conventional slide due to thick smear, air-drying artifact or
obscuring inflammation.
• Sharp, irregular cytoplasmic cavitation (HPV effect).
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Morphology I Slide: 76
LSIL
• This sheet of squamous cells exhibits well defined LSIL criteria:
nuclear enlargement, hyperchromasia, binucleation and cytoplasmic
cavitations
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Morphology I Slide: 77
LSIL
• These mature squamous cells show dark, hyperchromatic nuclei in
comparison to the normal cell population. Upon closer inspection, the
nuclear membranes exhibit irregularities.
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Morphology I Slide: 78
LSIL
HPV Effect
• Sharp, irregular cavitations in the cytoplasm are the most prominent
feature of these cells. Increased chromatin clumping is evident as is a
smudged nuclear appearance, secondary to HPV.
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Morphology I Slide: 79
LSIL
• On screening power it’s easy to see abnormal groups with
hyperchromasia, nuclear enlargement and HPV changes.
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Morphology I Slide: 80
Look-Alike: HPV Cavitation vs. Vacuoles
• Left image: Dense, sharp and irregular cavitations caused by viral
particles are evident. Cells infected with HPV have a “fried egg” (flat)
appearance as opposed to:
• Right image: which shows the smooth bordered glycogen vacuoles.
If present, the glycogen may have a waxy appearance. These cells have
a greater depth of focus with a bloated appearance.
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Morphology I Slide: 81
HSIL
• Sheets and syncytial groupings are maintained, not disrupted by the
dispersion process
• More distinct cytoplasmic borders are present and more information
can be gained from the cytoplasmic appearance
• Isolated immature cell forms present in background function as
“clue”
• Nuclear membrane irregularities are more prominent
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Morphology I Slide: 82
HSIL
• Metaplastic type squamous cells showing increased N/C ratios (in
excess of 50%), nuclear size and shape variability, nuclear membrane
irregularities and coarse chromatin.
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Morphology I Slide: 83
HSIL
• Dense immature cytoplasm, increased N/C ratios, and nuclear size
and shape variability result in a diagnosis of HSIL.
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Morphology I Slide: 84
HSIL
• At low power, abnormal single cells will serve as a “clue” and upon
further inspection, additional cells, both singly and in groups, will aid
in making the diagnosis.
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Morphology I Slide: 85
HSIL
• Upon closer inspection this small sheet of immature squamous cells
exhibit scant cytoplasm and hyperchromatic nuclei with coarse but
evenly distributed chromatin.
• These immature squamous cells are small and, during screening,
attention to the background cell population is necessary.
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Morphology I Slide: 86
Look-Alike: HSIL vs. Endometrials
• Left image: Groups of HSIL cells have less depth-of-focus, more
distinct cytoplasmic borders and greater nuclear variability.
• Right image: Endometrial cells present as 3 dimensional groups with
small, tightly packed nuclei that vary in shape but not size. The nuclear
features are consistent within the group and may frequently be
accompanied by menstrual blood.
Close inspection (on high power) is crucial to definitively identify
HSIL from benign endometrials.
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Morphology I Slide: 87
Look-Alike: HSIL vs. Atrophy
• Left image: Immature squamous metaplastic cells are present
exhibiting increased N/C ratios, hyperchromatic nuclei with nuclear
membrane irregularities and coarse chromatin.
• Right image: These parabasal cells show uniformity from nucleus to
nucleus as well as a bland chromatin pattern to aid in the determination
of a benign origin.
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Morphology I Slide: 88
Squamous Cell Carcinoma
• Tumor diathesis is maintained with the distinctive pattern of
proteinaceous debris, engulfed white blood cells, degenerated cellular
material and, in a case of keratinizing squamous cell carcinoma,
increased keratinized debris.
• Greater depth-of-focus to cell groups.
• Nuclei present with variability in chromatin pattern often with
parachromatin clearing.
• Nucleoli, if present, may be prominent.
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Morphology I Slide: 89
Squamous Cell Carcinoma
• Diathesis is present as necrotic debris in the background mixed with
protein, blood, and white blood cells. It is important to identify the
abnormal cellular component in the debris. Use the dirty/ratty
background as a contextual clue to alert you to the presence of disease.
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Morphology I Slide: 90
Squamous Cell Carcinoma
• Dense, angular cytoplasm helps to identify this group of malignant
cells as squamous in differentiation. Prominent nucleoli are present,
with irregular chromatin distribution and thickened nuclear
membranes.
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Morphology I Slide: 91
Squamous Cell Carcinoma
• Non-keratinizing squamous cell carcinoma can present with pale
immature cytoplasm. These nuclei have thickened and irregular
nuclear membranes and may appear pale but upon close inspection,
coarse chromatin and prominent nucleoli are present.
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Morphology I Slide: 92
Squamous Cell Carcinoma
• Single malignant cells may display dense cytoplasm with possible
orangeophilic staining. Nuclear detail is apparent with parachromatin
clearing, prominent nucleoli, and irregularities in the nuclear
membrane. It is essential to incorporate all criteria for an accurate
interpretation.
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Morphology I Slide: 93
Squamous Cell Carcinoma
• Isolated abnormal cell forms may be present, intermixed with
necrosis in the background. Caudate or elongate cells are maintained
during the dispersion process.
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Morphology I Slide: 94
Tumor Diathesis — Look-Alikes
• Cytolysis — stripped nuclei and cytoplasmic debris are present.
• Blood — lysed in the background with RBC ghost cells also evident.
• Trich — busy looking slide with organisms and bacteria present.
• Tumor Diathesis — debris tends to clump together with a frayed
edge. Malignant cells should also be in evidence.
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Morphology I Slide: 95
Look-Alike: Non-Keratinizing Squamous
Cell Carcinoma vs. Adenocarcinoma
• Left image: Non-keratinizing squamous cell carcinoma presents
predominantly with malignant cells singly and in disorganized,
irregular groupings. The abnormal cells around the edge of this group
exhibit irregular, hyperchromatic nuclei with coarse chromatin.
• Right image: Adenocarcinomas typically form 3 dimensional cell
clusters and may exhibit a “rounder” presentation due to the liquid
based fixation. The nuclei may scallop around the outside edge of the
group and display finely granular chromatin and prominent nucleoli.
Cytoplasmic vacuoles may be seen.
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Morphology I Slide: 96
Glandular Cell Abnormalities
• Revised Bethesda terminology for reporting abnormal glandular
cells.
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Morphology I Slide: 97
AIS
Same features as CP may be used for TP.
• Abnormal architectural patterns are maintained.
• Nuclear crowding with hyperchromasia.
• Pseudostratification and “feathering” are identified.
• Cell groups may present in rosettes and strips.
• Nucleoli may be present.
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Morphology I Slide: 98
AIS
• At screening power, numerous dark groups of cells should alert you
to a possible abnormal process.
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Morphology I Slide: 99
AIS
• The tissue pattern of AIS is maintained with nuclear stratification
and crowding, with nuclei falling outside of the grouping.
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Morphology I Slide: 100
AIS
• This group of glandular cells exhibits nuclear crowding as well as
hyperchromatic nuclei with a powdery chromatin pattern. Cell
dyshesion is evident along the edges.
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Morphology I Slide: 101
AIS
• This small group of cells shows pseudostratification with crowded,
hyperchromatic nuclei.
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Morphology I Slide: 102
AIS
• In AIS nucleoli may be present.
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Morphology I Slide: 103
Look-Alike: AIS vs. SIL in Glands
• Left image: AIS exhibits groups with a greater depth-of-focus and
nuclear crowding with finely granular or vacuolated cytoplasm.
• Right image: SIL appears flatter with a more sheet-like arrangement.
The cytoplasm is dense and the cells can have a lower N/C ratio.
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Morphology I Slide: 104
Adenocarcinoma
• 3D cell clusters maintained.
• Well preserved nuclear features are evident.
• Tumor diathesis may be noted with invasive glandular lesions.
.
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Morphology I Slide: 105
Endocervical Adenocarcinoma
• Cluster of abnormal glandular cells with scalloped edges and
irregular nuclei with finely granular chromatin and nucleoli.
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Morphology I Slide: 106
Endocervical Adenocarcinoma
• Loose cluster of large malignant cells exhibiting “rounding up.” Also
present are smooth cytoplasmic borders and nuclei with irregular
nuclear membranes and irregular distribution of chromatin.
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Morphology I Slide: 107
Endocervical Adenocarcinoma
• Loose cluster of malignant cells with vacuolated cytoplasm and
eccentric irregular nuclei.
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Morphology I Slide: 108
Endometrial Adenocarcinoma
• Papillary grouping with increased depth-of-focus and poly
engulfment.
• High N/C ratio and eccentric nuclei with nucleoli.
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Morphology I Slide: 109
Endometrial Adenocarcinoma
• Papillary cluster of uniform, bland appearing malignant epithelial
cells.
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Morphology I Slide: 110
Endometrial Adenocarcinoma
• Dense cluster of malignant cells with scalloped edges, vacuolated
cytoplasm, enlarged irregular nuclei with nucleoli and engulfed polys.
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Morphology I Slide: 111
Small Cell Undifferentiated Carcinoma
• Aggregates of small cells with hyperchromatic nuclei and nuclear
molding. Note the size compared to the neutrophil.
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Morphology I Slide: 112
Malignant Mixed Mesodermal Tumor
A: Adenocarcinoma component – a 3 dimensional cluster of tightly
packed cells showing poly engulfment.
B: Malignant spindle cell component – a loose aggregate of spindled
tumor cells with irregular nuclei and nucleoli. This component can be
of either squamous or mesenchymal origin.
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Morphology I Slide: 113
Malignant Melanoma, metastatic to cervix
• Numerous single malignant cells with irregular nuclei, prominent
nucleoli and some evidence of brown pigment in the cytoplasm.
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Morphology I Slide: 114
Malignant lymphoma, metastatic to cervix
• Necrotic debris with numerous single cells. On high power, nuclear
variability is evident and many nuclei appear with clefts and nucleoli.
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Morphology I Slide: 115
Part No. 86513-001 Rev. A
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Morphology I Slide: 116