Cytology Flashcards
What is a laboratory diagnosting test dependant on
Good history
Quality of sample
Proper identification of sample
How do you choose what sampling technique to use for histology
Anatomic location
Patient’s overall health
Suspected tumor type
Clinician’s preference
What are the pretreatment biopsy types
Needle core biopsy
Punch biopsy
Wedge biopsy
How do you obtain additional information about a tumor
treatment planning (surgical, medical)
What is excisional biopsy
Surgical removal of the tumor
what is a post treatment biopsy method
excisional biopsy
How do you obtain a more complete picture about a growth
Grading
Lymphatic/vascular invasion
Margins
Is a biopsy a good first step?
No
What is the disadvantage to not doing a biopsy first when a lump is removed
can result in incomplete removal, more morbidity and costs
What are the advantages to pre-treatment biopsies
Can help clients make an informed decision
Can consult with oncologist and surgeon
Can plan treatment sooner after surgery
When are pre-treatment biopsies not indicated
Treatment or Sx would not change (spleen, testicle)
As risky as removal (spinal cord)
What are needle core biopsies done on
external palpable masses (no highly inflamed or necrotic)
deep (kidney, liver)
Describe the needle core biopsy punch
manual or spring/pneumatic powered
Small sample size still enough for pathologic exam
what is the size of the needle core biopsies needle
1 mm wide biopsy
1.0 – 1.5 cm long
What does a needle core biopsy require
local anesthesia and sedation
sterile preparation
Why do you use a small scalpel incison for the needle core biopsy
Prevents dulling
Facilitates tru-cut mechanism
Can be sutured
How do you handle the tissue from the needle core biopsy
Tissue can be removed with blade, needle or saline
Can be rolled on glass slide for cytology
Place in formalin (in cassette)
What is a possible risk when you do a needle core biopsy
minimal risk of seeding but you should plan ahead and remove original incision tract
consider hemorrhage and fluid leakage
Why do you use a punch biopsy
Typically for skin
Skin, oral, perianal
Direct access with laparoscopy
Liver, GIT, etc.
What is the size of a punch biopsy
2-8mm
What is required for punch biopsy
local anesthesia and sedation
usually no sterile preparation
what is the ideal size of a punch biopsy
6mm
4 mm only for nose, footpad
8 mm slight more chances of infection
What can cause tissue compression and artifacts when doing a punch biopsy
dull punches
how do you handle a tissue sample from a punch biopsy
handle sample very gently
place in formalin, no cassette
what is important for punch biopsies if you’re doing dermatology
draw line in direction of hair
When do you do an incisional biopsy
When cytology and/or biopsy is unsuccessful
For ulcerated and necrotic lesions (larger sample)
What do you need to do for an incisional biopsy
Surgical preparation + drapes
Local anesthesia
Tumors are usually POORLY innervated
Skin is incised and tumor wedge removed
Is it necessary to remove intact skin with the incisional biopsy?
NOT necessary to remove intact skin (next or over)
Margins evaluated with removal of tumor
Can compromise
Careful not to sample just the reactive tissue surrounding the tumor
Imprint cytology can be done
Describe endoscopic biopsy
Convenient, cost-effective, safe
Limited sample, inadequate visualization
Describe laparoscopy, thoracoscopy
Very good, can always convert to laparotomy
Needs specialized tools & skills
How do you properly identify margins
tissue ink is preferred but sutures can also be used.
need to write: color = which margin.
when should tissue ink be done
before fixation, to orient the sample and identify areas of concern
describe the tissue inking process
Tissue should be blotted with paper towel before
Ink can be applied with gauze on surface or cotton swab for precision
Allow to dry 20 min before formalin
describe proper tissue fixation
10% buffered neutral formalin
Special fixative for eyes, testicles
1 part tissue to 10 part fixative
Ideally 1 container per lesion
done within 30 minutes
describe containers for fixation
Wide container, secure lid No glass No more than 1L In secure plastic bag (ziploc) With absorbent packing material Insulated (avoid freezing)
Descriibe large unusual specimen fixation
Amputated limbs, spleen
Can overnight on ice
Can pre-fix 48-72h with partial parallel incisions approximately 1 cm apart (‘‘bread loafing’’)
Then shipped without formalin (double-bag) or in 1:1 formalin
Can section and fix/send in individual containers
An annotated digital image or sketch of the original specimen to depict sectioning and orientation should accompany the samples
describe very small unusual specimen fixation
Labeled cassettes
Do not use gauze sponges or cardboard because tissue may become compromised upon retrieval
describe luminal organs unusual specimen fixation
Flush the intact lumen with formalin
Partial longitudinal incision
3 labeled sections (cranial/proximal, mass and caudal/distal) can be submitted
describe thin flat sample fixation
Small: placed in a tissue cassette with a foam pad to minimize tissue curling
Larger samples can be tacked onto a flat piece of cardboard presoaked in formalin or water with suture through edges of tissue not needed for examination
What do you need to write about the lesion in the mass submission form
Lesion-specific clinical history (eg, anatomic site, date first noticed, rate of growth)
Potentially lesion-associated clinical signs (eg, lameness, vomiting)
Type of lesion (eg, new lesion, recut following incomplete excision, excisional biopsy following previous incisional biopsy, local recurrence)
Results of prior lesion-associated diagnostic tests - cytology, prior biopsy reports, imaging (radiographs, ultrasound, MRI, CT); access to radiographs may be especially important for bone and gingival tumors.
What general information do you need to put on the mass submission form
General clinical history—previous neoplastic diseases, previous or current nonneoplastic conditions of relevance
Treatment history—local and systemic, current and previous (eg, chemotherapy, radiation, corticosteroids)
Previous unrelated treatments or potential tumor-inducing historical events at tumor site (eg, previous radiation, vaccination, implants)
What other abnormalities should you include on the mass submission form
CBC, biochemical, and hormonal (eg, hyperinsulinemia) abnormalities
What else should be on the submission form
Working clinical diagnosis and/or list of differentials
Thorough gross lesion description.
Indication whether the submitted sample is an incisional or excisional biopsy.
Excisional biopsy indicates assessment of surgical margins is necessary, whereas for incisional biopsies the margin evaluation is null.
Anatomic site should be thoroughly described
Features appreciated during diagnostic imaging or perioperatively should be described
tissues involved or associated with the mass (eg, thyroid mass invading subjacent skeletal muscle).
What is cytology
examination of cells having exfoliated from tissue or having accumulated in body fluid
what can cytology provide
May provide definitive diagnosis
Very dependent on sample quality!
is cytology invasive?
Not invasive
How quickly does cytology need to be processed
rapidly
What should be in the cytology kit
Clippers
Cleansing & disinfectant wipes
Syringes: 6 – 12 ml, up to 20 ml
Needles: 1 – 1.5 in (20g to 22g), 2.5 – 3.5 in spinal needle with stylet
Bone Marrow aspiration needle & core biopsy material
Scalpel blades
Culture swabs & applicator sticks for slide preparation
Box of slides (frosted)
EDTA and red top tubes
Rigid, flat surface for 6 -10 slides (foam tray)
Butterfly catheter, IV extension tubing
Pencil or slide marker
Sterile EDTA
What is the skin prep for a FNA
Minimal for cutaneous, subcutaneous
Shave, clean & disinfect for internal
Where can you do an FNA
Cutaneous, subcutaneous, internal organs
What are the benefits of going needle only, vs aspiration
Should start with no aspiration
Less blood contamination
What is the best FNA size needle
22g
What do you do if a FNA sample is liquid
place in edta
what do you do if an FNA sample is solid
prepare slides immediately
Describe imaging guided aspiration
typically ultrasound guided
complications are rare
thoracic samples can be taken.
Should ascites and pleural effusion be sampled via US
yes
Describe the squash preparation
Most common
For semi-solid, mucus-like, or pelleted (via centrifugation)
Place sample close to frosted edge and use second slide to spread sample
What do you do with fluid cytology samples
Keep in EDTA Prevents clotting (fibrin) Preserves cellular morphology Facilitates cell count Refrigerated Up to 24h in general CSF needs special measures
Make slide as soon as possible
Send unstained slide with EDTA tube
How do you prepare a fluid slide if the sample is cloudy
direct smear (like blood smear) squash
how do you prepare a fluid slide if the sample is clear
If lower cellularity Need to concentrate Similar to urine sediment Via special centrifuge (CSF, BAL) Buffy Coat Still do a direct smear
What hematology methods can you use with cytology fluid
Cell counts Can be automated (hematology machine) Flush with saline after Manually Total proteins Refractometer Conversion tables for low TP
Describe the touch imprint
Permits evaluation of a biopsy
With surface lesions is often of poor diagnostic utility
Superficial inflammation, secondary bacterial infection
Exception for fungal diseases
Need to blot aggressively the sample Until tacky Then imprint on slide Fibrous lesions can be scraped Scalpel blade
How do you prepare joint-synovial fluid cytology
Surgical preparation
Normally viscous
Ideally small EDTA tube To get cell count To be able to use hyaluronidaze Direct smear (like seen previously) Culturette for microbiology
What are miscellaneous types for doing cytology
Always air dry – do not use flame Do not freeze Do not expose to formalin Label properly all tubes & slides Always submit an unstained smear with a tube (also for hematology)
When do you do bone marrow evaluation
Bone marrow evaluation is indicated when peripheral blood abnormalities are detected
persistent neutropenia, unexplained thrombocytopenia, poorly regenerative anemia
To stage neoplasia
Lymphoma, plasma cell tumor, mast cell tumors, other
What is the difference between aspiration and core biopsy
Aspirates are easier, faster, and less expensive to perform than are core biopsies.
Bone marrow core biopsies require special needles.
Core biopsy sections provide a more accurate way of evaluating marrow cellularity and examining for metastatic neoplasia than do aspirate smears, but cell morphology is more diffcult to assess.
How can you classify inflammation
purulent pyogranulomatous macrophagic eosinophilic lymphocytic
Describe purulent inflammation
predominance of neutrophils (>85%)
Try to say if degenerated or not
Describe pyogranulomatous inflammation
mix of neutrophils and macrophages
Describe macrophagic inflammation
predominance of macrophages (>50%)
Describe eosinophilic inflammation
important component of eosinophils (>10 – 30%)
describe lymphocytic inflammation
need to rule out lymphoma
What does it mean if you have degenerate neutrophils in your purulent inflammation
Degenerate neutrophils:
Bacterial infections
Need to see to call septic
What are the causes of degenerate neutrophils in purulent inflammation
Degenerate neutrophils:
Immune-mediated
Neoplastic
Sterile irritants (bile, urine)
What are granulomatous lymphocytes associated with
Foreign body Fungal infection Mycobacterial infection Panniculitis Lick granuloma Other chronic lesions
what is eosinophilic inflammation due to
Eosinophilic granulomas Hypersensitivity Parasites Fungal Mast cell tumors Some neoplasms
what is lymphocytic inflammation due to
Rare
Immune reaction
Viral
Chronic lesion
What is the infectious agent blastomycosis associated with
Pyogranulomatous or granulomatous Dogs mostly “hunting” Nose, legs Found in the environment
What is the infectious agent cryptococcus associated with
Granulomatous
Dogs & Cats
Nose
where is the aspergillus fungi found
Opportunistic
Dog: nose
Horse: cornea
Describe mycobacterium
Fairly rare
Granulomatous
Very slow to grow in microbiology
How do you classify a neoplasm
need to say the cell type
if it is benign or malignant
what are the 4 cell types of neoplasms
epithelial cells
mesenchymal cells
round cells
neuroendocrine cells
how do you classify if a cell is benign or malignant
Set of criterias
Cytoplasmic
Nuclear
what are the cellular features of malignancy
Cellular Crowding Pleomorphism Different shapes Anisocytosis Different cell size Giant cells Basophilia
High N/C ratio (nuclear/cytoplasmic)
Not always
what are the nuclear features of malignancy
Nuclear Nuclear molding More than 1 Pleomorphisme Anisocaryosis Within a cell also Mitotic figure Number and shape
what are the nucleolus features of malignancy
multiple
varied within one nucleus
may be normal
Describe the organization of mesenchymal cells
Weak cohesion, loosely arranged
Often extra-cellular matrix
describe the cellular types of mesenchymal cells
Cellular types
Eg. fibroblasts, osteoblasts, chondroblasts…
describe the morphology of mesenchymal cells
Morphology
Spindled, stellate, oval
Poorly defined cytoplasmic margins
describe the exfoliation for mesenchymal cells
Exfoliation
Moderate to weak
describe the organization of epithelial tumors
Cohesive clusters
describe the cell types of epithelial tumors
Glandular and parenchymal tissue
Surface lining
Eg. Basal cells, squamous cells, hepatocytes, tubular cells, renal cells
describe the morphology of epithelial tumors
Variable: round to polygonal +/- elongated
Distinct cytoplasmic borders
describe the exfoliation of epithelial tumors
very easy
What are all the round cell types
Lymphome
Plasmocytome
Mastocytome
Histiocyte
Sarcome histiocytaire
Transmissible veneral tumor
What does it mean when you see lymphoglandular bodies
Cytoplasm fragment
Mostly seen with lymphocytes
lymphoma
what does it mean when you see collagen breakdown
Mostly seen in mast cell tumors
Some soft tissue sarcoma
what does it mean when you see a hematoidin crystal
Hematoidin crystal
Hemoglobin breakdown
Indicates chronic bleeding
what does it mean when you see cholesterol crystals
Cholesterol crystals
Cell membrane damage
Frequent in
Follicular cysts
what does it mean when you see skeletal muscle
Skeletal muscle
Normal
Incidental finding
