cytology Flashcards
cytology
Study of cells Exfoliative=cells shed from body
fine needle aspiration
- Hold the mass firmly
- stick mass with needle and syringe
- draw back on the plunger until about three fourths of the volume of the syringe
- Redirect the needle to collect sample regions of the mass—maintaining negative pressure
- Remove from mass after removing negative pressure and remove the needle from the syringe
- Draw air into syringe and replace needle
- Expel needle contents onto slide
direct imprints
- Imprint before the lesion is cleaned
- Then clean the lesion with saline-moistened surgical sponge Imprint No. 2 is taken
- Debride lesion and imprint No. 3
- If a scab is present, imprint underside of scab No. 4
- Imprint exposed tissue No. 5
fluids and washes
- 18- or 20- gauge through-the-needle catheter
- Cliphair of laryngeal area and aseptically prepare Lidocaine for local anesthesia
- Needle inserted into trachea through cricothyroid membrane and catheter advanced into the lumen of the trachea
- Sterile saline is infused through catheter
- When animal coughs, pull back plunger to collect fluid
swab
moisten sterile cotton swab with sterile saline, after collection gently roll swab along slide do not rub
methods for properly preparing a slide from FNA, swabbing, fluids and washes, and solid tumor
Compression, Combination, Starfish, Line Smear, Wedge smear
stains of choice
Romanowsky stains (Wright, Giemsa, DiffQuick, DipStat), New Methylene Blue
how to examine a cytological slide
Start at 10X to find islands of cells then switch to 40X; Yes coverslip if no oil immersion, if oil immersion no coverslip
how samples should be prepared for sending to a referral lab. If you look at the slides prior to sending them, what must you NOT do?
- 2-3 air dried unstained slides
- 2-3 Romanowsky stained slides
- direct smears and EDTA and Red top tubes
- protect slides, DO NOT SEND SLIDES WITH OIL
parts of the body from which cells may be collected for analysis and the method that may be used to collect cells from each area
- Bodily fluids- cerebrospinal, peritoneal, pleural, synovial (washes and aspiration)
- Mucosal surfaces-trachea, vagina (transtracheal wash, percutaneous, swabbing)
- Secretions-semen, prostatic fluid, milk (collection into proper tube, endoscopic method, FNA)
how a sample is collected from a dog and prepared for vaginal cytology examination
Swab with cotton swab moistened with sterile saline, roll onto slide
Karyorrhexis
Cell fragmentation
Karyolysis
Rapid cell death
pyknosis
Slow cell death (aging)
degenerative neutrophil
swelling of the nucleus, nucleus appears lighter stained and smudged
sepsis
infection
non - degenerative neutrophil
these are neutrophils with tightly clumped, basophilic nuclear chromatin
Neoplasia
Contain a homogeneous population of a single cell type
Macrophage
tissue resident immune cells and can be generated from monocytes in the blood after their activation and tissue infiltration
Inflammatory
characterized by WBCs
Malignant
Cancerous cells likely to spread and cause more serious disease
Benign
cells that are not likely to spread to other areas
nucleolus/nucleoli
Malignant cells display abnormal configurations
procedure for initial examination a cytology slide
10X objective: estimate 2 dominant cell types by scanning multiple fields, switch to 40X identify multiple fields one at a time
As you first examine a slide, what 3 things should you look for that would prompt you to make a new slide?
Smear is too thick, has few cells, or mostly broken cells
What do you determine first when looking at the cells?
Identify dominant cells, and decide whether the sample is non-inflammatory or inflammatory
What cells are dominant in an inflammatory response?
Neutrophils,macrophages, or eosinophils
3 types of cells seen if there is neoplasia
Round cells
Mesenchymal (spindle cells)
Epithelial cells
round cells
individualized, round nuclei, distinct cell borders, high cellularity
mesenchymal spindle cells
wispy, frayed, indistinct cytoplasmic borders, low to moderate cellularity, disorganized clusters
epithelial cells
Polygonal, caudate, or round. (like epithelial types in urine) Distinct cell borders. “Organized” clusters. Moderated to high cellularity
characteristics of a neoplastic cell in comparison to benign cells
- large variations in size, appearance
nuclei tend to be large, and they may have course chromatin - there may be more than one nucleus in a cell or other signs of abnormal mitosis nucleoli are present in irregular numbers and sizes
- large variations in the ratio of nucleus size to cytoplasm, cytoplasm may be very basophilic, cell margins may be frayed, Purple granules or black and green granules in the cytoplasm of cells with round nuclei
Neutrophils and macrophages are the most common cells seen in inflammation caused by fungus or
parasites
What other cell types indicate inflammation related to parasites or allergy?
eosinophils
How can you tell mature lymphocytes from neoplastic round cells on a cytology smear?
neoplastic cells will display at least three abnormal configurations within the nucleus such as nucleoli with variable shapes, a diffuse chromatin pattern, etc.
5 characteristics that a degenerative neutrophil might exhibit
karyolysis, karyorrhexis, pyknosis, swollen cell, toxic granules, frothy cytoplasm, and/ or increased basophilia of cytoplasm
Sepsis is likely when we see large numbers of _____. If we see cells of these types with bacteria in the cytoplasm, that will confirm it.
degenerate neutrophils
What else can cause inflammation other than sepsis?
Non-septic inflammation Indicates the body is trying to get rid of dead tissue, a foreign body, or abnormal tissue growth (neoplasia)
Describe a mast cell. Which of the 3 types of neoplastic cell types is it? How can it be differentiated from a basophil? How can it be differentiated from melanoma?
Mast cell tumors—prominent purple/black granules (round cells - no indented nucleus like baso)
Melanoma—cells with prominent dark black granules
You see a bunch of strings of blue on a cytologic smear and they all go the same direction. What are these? What is the cause?
Smudged cells from incorrect technique
What is the name of the yeast common in ear infections? What does it look like?
Malassezia, footprint
Besides yeast, what else would you look for on an ear smear that would indicate infection?
bacteria
You are looking at a slide made from swabbing out an ear. You see cellular debris and 3 yeast cells and 10 rod bacteria on the whole slide. Should you tell the vet that the slide indicates the animal has an ear infection?
This would be few yeast and few bacteria so no
You rolled a swab of material taken from the ear canal on a slide. What do you do to the slide before staining it?
Heating the slide can help get rid of excess cerumen
If you stain a slide made from an FNA of a lipoma with Sudan stain, what will the cells look like? What is a lipoma? If you stained it with Dif-Quick, what would you see on the slide?
Sample looks like oil on the slide; Sample disappears after fixed in Dif-Quick. Staining slide with NMB or Sudan IV stain shows outline of large, fat filled cells with dot-like nuclei near edge.
microscope settings for vaginal cytology
40x, medium light
why is vaginal cytology not done on cats
potential ovulate because they think they’ve been mated
4 phases of canine estrous cycle
anestrus
proestrus
estrus
diestrus
anestrus
- no vulvar swelling
- doesn’t attract male
- primarily parabasal and small intermediate
proestrus
- attracted to male
- first heat cycle
- leaning off paranasal to more intermediate and superficial
estrus
- accepts male
- mainly anclear
diestrus
- decrease in swelling
- no longer attracts male
- anuclear disappears and parabasal reappears
two reasons besides breeding status for doing a vaginal cytology
vaginitis and metritis
vaginitis
inflammation of vulva
metritis
inflammation of uterus
