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Define cytology.
removing cells from a tissue and examining them microscopically.
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What is the alternative to histopathology?
cytology
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Is performing a cytology expensive?
no
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What is cytology used to identify?
- masses
- lesions
- fluid in body cavity
- pathology of internal organs
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What is a lesion?
an abnormality in the tissue
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What are the advantages to doing a cytology?
- no extra equipment needed
- very quick procedure
- relatively non-invasive (animal is usually awake)
- often can ID cellular reaction
- may be able to ID cause
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What are the disadvantages to doing a cytology?
- some training is needed to ID cells present
- may not be able to ID cells
- may need to supplement with histopathology
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Why would we not be able to ID cells in a cytology and what does this mean?
- cells could be unrecognizable
- tells us something is wrong, but not what is wrong - usually means neoplasia of some sort
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Who is able to perform a cytology and read a cytology?
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What is required to do a histopathology?
a tissue sample
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How do we do a histopathology?
- anesthesia is often required
- surgically remove the sample of tissue (sometimes its just a part of the tissue or the whole thing)
- fixed it in 10% buffered formalin
- send to pathologist
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How much 10% buffered formalin is needed for a histopathology?
use 10x amount of formalin to size of the tissue sample
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How thick does the tissue sample need to be in order for the 10% buffered formalin to fix it?
1 cm or less
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Once we send the histopathology sample off to the lab, what happens?
- gets dehydrated
- set in parifin (wax)
- sliced
- mounted on a slide
- stained in HNE (blue and red stains)
- analyzed by a pathologist
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What are the advantages to doing a histopathology?
- slide examination is done by a trained pathologist (veterinarian with advanced training)
- tissue architecture around cells are visualized
- special stains can be used
- may be given a more precise diagnosis (especially with neoplasia)
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What are the disadvantages to doing a histopathology?
- more involved procedure
- more expensive to the client
- pathologist is unable to see the patient and can be fooled if it is a poor sample or the information is not thorough enough
- results take time
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What are the different cytology techniques?
- aspiration
- impression smear
- wash
- scraping
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If we think there is a presence of bacteria should we culture it first before doing the cytology?
yes
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What is another name for an aspiration?
FNA (fine needle aspiration)
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What materials do we need to do an aspiration?
- 22 - 25g needle
- 3 - 12 cc syringe (don't always need to use)
- slides
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How do we do an aspiration with a needle and syringe?
- insert needle and syringe
- aspirate
- redirect needle and reaspirate
- stop once you see tissue into the hub
- remove needle and pull back the syringe to get air
- put the needle back on the syringe and push the air out onto a slide
- the sample from in the hub will come out onto the slide - flea sneeze
- lay another slide at 90 degrees and pull apart
- make at least 2 slides
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The sample we get from an a FNA is very small, what do we call it?
flea sneeze
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What is the way to do an aspiration without using a syringe to aspirate?
- just stick the needle directly into the lesion
- redirect the needle several times
- take the needle out and attach it to syringe that has air in it
- push the air out onto a slide (flea sneeze)
- lay another slide at 90 degrees and pull apart
- make at least 2 slides
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If we are doing a centesis, should we stop aspirating when we get the sample in the hub of the needle?
no keep pulling back on the syringe to collect the fluid into the syringe
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How do we do an impression smear?
- avoid cleaning the lesion with anything first (can use alcohol if it is really dirty)
- blot blood off the lesion first (don't wipe it off, just blot)
- gently (avoid pressure) touch slide to the lesion
- can touch in several different places for each slide
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What if we want to do an impression smear but the lesion is in a place we can't get to with a slide?
- use a sterile swab
- moisten with saline if site is dry
- blot blood first
- roll swab across lesion then across the slide
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How do we do a wash or a flush?
- flush lesion with saline
- aspirate back on syringe
- may use a catheter to get to the sample and attach a syringe on the end
- smear sample onto slide or centrifuge the sample down and then smear it onto the slide
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When would we use a scraping?
used with firm lesions
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How do we do a scraping?
- hold blade at 90 degrees to surface of the skin
- scrape back and forth
- spread debris on slide
- lay another slide on top and pull apart
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Should we stain/fix both of the slides we make? Why or why not?
no, only stain 1 because if we see something on the slide that needs to send out to the lab, the lab does not want it stained
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Which types of stains can we use?
- romanowsky (diff quik)
- new methylene blue (to look at nuclear morphology)
- gram stain (for bacteria)
- special stains
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Gram positive organisms stain _____ with gram stain.
blue
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Gram negative organisms stain _____ with gram stain.
red
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What are other special stains that can be used?
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Where can we get special stains?
almost all human hospitals have the special stains
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Once we have stained our cytology, how do we examine the slide?
- look with low power to evaluate staining, cellularity
- move to high power
- only use oil if we can't see what we need on high power
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What are we looking for on the slide?
- cellular reaction occuring
- cause of the reaction
- don't ignore bacteria, fungus, or parasites
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What types of cellular reactions could occur?
- inflammation
- hyperplasia
- neoplasia
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What is an inflammatory reaction?
body's reaction to some insult
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What are the 3 different types of inflammatory reactions?
- acute
- chronic active (or mixed inflammatory reaction)
- chronic
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What are the 4 cardinal signs of inflammation when looking at an animal?
- swelling
- redness
- warmth
- pain
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Does inflammation equal infection?
no
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What is acute inflammation?
>70% of inflammatory cells are segs
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Could cells other than segs be present in acute inflammation?
yes, don't ignore them because neoplasia can be associated with inflammation
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What types of segs could we see with acute inflammation?
- normal segs
- hypersegs
- degenerating segs
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When we are looking at a cytology, do we need to count the cells?
no, just scan
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If we want to see bacteria what magnification do we need to be on?
oil
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What are the different characteristics of degenerating segs?
- pyknosis
- karyorrhexis
- karyolysis
- cytoplasmic basophilia
- vacuoles
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What is pyknosis?
small, condensed, dark nucleus
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What is karyorrhexis?
nucleus explodes and we see different pieces of the nucleus inside and outside the cell
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What is karyolysis?
- swelling of nuclear membrane and cell gradually loses chromatin
- nucleus is "melting away"
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Are there a lot of eosinophils in acute inflammation?
no
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If we see >10% of eosinophils in acute inflammation then what is it called and what is it due to?
- eosinophilic inflammatory reaction
- often due to allergy, parasites, eosinophilic disease
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What is chronic active inflammation?
50 - 70% of cells are segs and the remainder are monocytes and macrophages
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What is another name for chronic active inflammation?
mixed inflammatory reaction
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What is chronic inflammation?
more than 50% of cells are monocytes and macrophages
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When will we see chronic inflammation?
often seen with systemic fungal infections and foreign bodies
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What is granulomatous inflammation?
a form of chronic inflammation
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What normally happens with granulomatous inflammation?
- fibrous wall is built around the lesion (firm small or large white masses)
- bodys response to the organism in the body
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What are special macrophages?
- epithelioid cells
- giant cell
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What can special macrophages be confused with?
neoplasia
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What does an epithelioid cell look like?
- large cell with an abundance of pale cytoplasm
- vesicular nucleus
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What does a giant cell look like?
- large, multinucleated cell
- an abundance of pale cytoplasm
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What is another way to look at a slide on oil and why is it better?
- put oil on the slide and put a cover slip over it
- examine on 400x
- increases clarity and resolution
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To find organisms in a cytology where do we need to look?
- in segs or between cells
- check in macrophages too
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What color does mucus stain?
pink
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Should we look at discharges under a microscope?
- yes
- make 2 smears
- air dry and stain one
- may need specific stains
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What are the specific stains we may need for examining discharges?
- gram stain (for bacteria and yeast)
- PAS (makes fungal organisms stand out)
- acid fast stains (for acid fast organisms)
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What are the different types of tissue we can examine on a cytology?
- epithelial
- connective
- muscle
- nervous
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What are some types of epithelial tissues?
- squamous (in body openings)
- cuboidal (in glands)
- columnar (in intestines and lungs)
- glandular
- transitional (in bladder and urinary tract)
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What are some types of connective tissue?
- bone
- cartilage
- fibrous
- blood
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What is hyperplasia?
- tissue enlarged or thickened
- increased population of normal cells
- normal body process
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Which tissues would we see hyperplasia?
- lymph nodes
- spleen
- prostate
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What does a cytology look like of hyperplasia?
- mostly young cells
- large nuclei with vesicular chromatin
- basophilic cytoplasm
- high N:C ratio
- normal mitotic figures (chromosomes line up like they normally do in mitosis)
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What kind of inflammation is this?
acute inflammation (>70% inflammatory cells are segs)
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This urine sample was collected via cystocentesis. What is wrong?
- too much bacteria for a cysto collection
- indicates cystitis
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What is wrong with this fecal sample?
camplyobacter
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What kind of inflammation is this?
chronic active inflammation (50 - 70% inflammatory cells are segs and the rest are monocytes and macrophages)
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What kind of inflammation is this?
chronic inflammation (>50% inflammatory cells are monocytes and machrophages)
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What is wrong with this fecal sample?
clostridium
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What type of inflammation is this?
eosinophilic inflammatory reaction
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What is wrong with this fecal sample?
giardia
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What kind of stain was used for this?
gram stain
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hyphae from nasal discharge
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What kind of smear technique is this?
impression smear
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What kind of stain was used?
new methylene blue
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What kind of organism is this?
nocardia (seen more in large animals)
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What is this? It is from the prostate.
prostatic hyperplasia
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What is wrong with this seg?
pyknosis
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What is wrong with these cells?
pyknosis
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What kind of cytology technique is this?
aspiration (redirecting)
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What kind of cytology collection technique is this?
scraping
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What kind of organism is this?
simonsiella (normal flora of the mouth)
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What kind of sample collection is this?
wash
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What kind of inflammation is this?
granulomatous inflammation (part of chronic inflammation)
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epithelioid macrophages (large cell, abundance of pale cytoplasm, vesicular nucleus)
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What are the 3 different types of cellular reaction?
- inflammation
- hyperplasia
- neoplasia
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What are the two types of neoplasia?
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We classify the neoplasia into benign or malignant based on what?
biological behavior
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Describe benign neoplasms.
- circumscribed
- grow slow
- not invasive
- grow to a certain size then stop
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What is circumscribed?
distinct nodule all the way around the mass
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If there is a benign neoplasm in a gland what might happen to the secretions of the gland?
they might increase
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What are some common benign tumors?
- fibroma
- lipoma
- adenomas
- skin tumors (melanoma, basal cell tumor)
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What do we call a fibroma in horses?
sarcoid
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What does a fibroma feel like?
firm
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What is a lipoma?
fatty tumor
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Where do we find adenomas?
glands - mammary, adrenal, thyroid
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What is the difference between neoplasm and neoplasia?
- neoplasm is a mass/tumor
- neoplasia doesn't necessarily have to have a mass or tumor
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Do malignant neoplasms grow fast or slow?
grow fast
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Are malignant neoplasms locally invasive/destructive?
yes, they do not have a nice edge around them like benign neoplasms
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Do malignant neoplasms metastasize?
yes
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What does metastasize mean?
spread to distant places
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Where do malignant neoplasms usually metastasize to first?
usually go to the lymph nodes first and then to the lungs
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Malignant neoplasms are anaplastic. What does this mean?
all the cells look like large immature cells and you can't tell them apart (undifferentiated)
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What do we analzye for the cytology of neoplasia?
nuclear and cytologic criteria
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What is pleomorphism?
variability in cell size and shape
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The more present the nuclear and cytologic criteria, the more _____.
malignant
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What is the nuclear criteria for determining malignancy?
- multiple number and sized nucleoli
- multiple number and sized nuclei
- variable staining of chromatin
- irregular nuclear membrane
- variable N:C ratio from cell to cell
- abnormal mitotic figures
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What is anisokaryosis?
difference in size of nucleus from cell to cell
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What is the cytoplasmic criteria for determining malignancy?
- increased basophilia (bluer cytoplasm)
- increased vacuolation
- varying amounts of cytoplasm per cell
- indistinct cytoplasmic edges (hard to tell where once cell stops and the next one starts)
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What are the different types of malignant tumors?
- carinoma
- sarcoma
- discrete cell neoplasms
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Where are carcinomas?
- epithelial tissue
- glands (adenocarinoma)
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Where are sarcomas?
connective tissue
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What are discrete cell neoplasms?
not just limited to one tissue type
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Describe sarcomas.
- firm
- few cells seen
- cells elongated - hard to tell one from the other
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Because sarcomas are firm, how do we make the cytology?
scrape
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What are some common sarcomas?
- osteosarcoma
- fibrosarcoma
- lymphosarcoma
- chondrosarcoma
- hemangiosarcoma
- melanosarcoma
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Are carcinomas very cellular?
yes
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What are some common carcinomas?
- squamous cell carcinoma
- mammary adenocarinoma
- prostatic carcinoma
- adrenal adenocarinoma
- thyroid adenocarinoma
- transitional cell tumor
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What are some different discrete cell tumors?
- melanoma
- mast cell tumor
- lymphoma
- histiocytoma
- transmissible venereal tumor
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What is the most common skin tumor in cats?
basal cell tumor
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Are basal cell tumors usually benign or malignant?
benign
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Where do we usually see basal cell tumors?
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What do basal cell tumor cells look like?
- cells clump
- high N:C ratio
- blue cytoplasm
- may see melanin and vacuoles
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What are plasmacytomas?
plasma cell tumors
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Which species do plasmacytomas usually affect?
dogs
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Where on dogs are plasmacytomas commonly seen?
- oral cavity
- ears
- forelimb skin
- digits
- GIT
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Are plasmacytomas usually benign or malignant?
benign
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When are plasmacytomas malignant?
when they are in the marrow
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What are melanomas?
neoplasia of melanocytes
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Where are melanomas common?
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_____% of melanomas on skin are _____.
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Most melanomas in the mouth and eyes are _____.
malignant
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What do melanoma cells look like?
brown to green-black granules that may obsure the nucleus
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Are melanomas common in cats?
no
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What is the most common skin tumor in dogs?
mast cell tumors
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What is the second most common skin tumor in cats?
mast cell tumor
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Are all mast cell tumors potentially malignant in dogs?
yes and we should treat all of them this way
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Mast cell tumors have an increased chance of being malignant if they are seen around what parts of the body?
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What are histiocytomas?
benign cutaneous tumor
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What do histiocytomas look like?
usually round nuclei with moderate basophilic cytoplasm
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What are histiocytomas often confused with?
lymphoma
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What age dogs do we usually see histiocytomas in?
< 3 years old
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Are histiocytomas fast or slow growing?
fast growing
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Can histiocytomas get better on their own?
yes
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Where are transmissible venereal tumors?
on the genitalia or on the lips
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How do dogs get transmissible venereal tumors?
from other dogs
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What do transmissible venereal tumors look like?
- numerous round cells
- abundant blue cytoplasm
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What type of nuclear criteria is this representing?
abnormal mitotic figures
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basal cell tumor on a cat
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What type of neoplasia is this?
carcinoma
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What type of sarcoma is this?
chrondrosarcoma (in the cartildge)
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What type of nuclear criteria is this representing?
variable staining of chromatin
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What type of cytoplasmic criteria is this representing?
cytoplasmic vacuoles
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What type of cell is this?
a glandular cell
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What type of sarcoma is this?
- hemangiosarcoma - tumors in blood vessles in the spleen and liver
- this is of the spleen
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What type of sarcoma is this?
hemangiosarcoma
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