Cytology 1 and 2
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What is the primary purpose of exfoliative vaginal cytology?

  • To diagnose otitis externa
  • To collect samples for fecal cytology
  • To detect bacterial and fungal infections
  • To determine the stage of the estrous cycle in bitches and queens (correct)

    • What is the first step in obtaining a vaginal cytology sample?

  • Introduce the lubricated speculum into the vagina
  • Rotate the swab
  • Clean and rinse the vulva (correct)
  • Stain the slides with diff quik

    • What is the characteristic of vaginal epithelial cells during estrus?

  • They become smaller and more irregular
  • They disappear
  • They become large and flattened with small or absent nuclei (correct)
  • They remain unchanged

    • What is the purpose of tracking changes in the morphology of desquamated vaginal epithelial cells?

    <p>To monitor changes in estrogen levels</p> Signup and view all the answers

    What is the name of the stage of estrus characterized by a swollen vulva, reddish discharge, bitch will attract but not accept male in attempts to breed

    <p>Proestrus</p> Signup and view all the answers

    What is the purpose of staining the slides with diff quik?

    <p>To highlight cellular morphology</p> Signup and view all the answers

    What stage of estrus do you suspect if you see noncornified squamous epithelium, intermediate or parabasal epithelial cells, neutrophils (may be) present in small numbers, and RBCs are seen

    <p>Anestrus</p> Signup and view all the answers

    What is the primary focus of fecal cytology?

    <p>Analyzing fecal samples for bacterial and fungal infections</p> Signup and view all the answers

    What stage of estrus do you suspect when the animal has a history of proestrus, discharge has changed from red to pinkish (and becoming whiter in the late stages), the bitch accepts males and microscopically squamous epithelium is cornified (there are also neutrophils and RBCs are present but decreased)

    <p>Estrus</p> Signup and view all the answers

    What stage of estrus do you suspect if vulvar swelling and discharge have decreased, the bitch no longer attracts males, cornified epithelium is replaced by non cornified squamous epithelium, neutrophils are present again and RBC are absent

    <p>Diestrus</p> Signup and view all the answers

    Which stage of estrus (in canine) is longest, lasting 60 days, without vulvar discharge, predominant cell is noncornified epithelium and progesterone secretion is at its peak

    <p>Diestrus</p> Signup and view all the answers

    What is the primary reason for collecting multiple swabs during the examination of the external ear canal?

    <p>To examine for parasites</p> Signup and view all the answers

    The onset of diestrus is marked by a precipitous decline in the number of superficial cells and reappearance of intermediate and parabasal cells. Most commonly, the cellular profile changes within a single day from essentially 100% superficial cells to less than 20%. However, It is best to confirm the onset of diestrus by examining a smear prepared of diestrus day __

    <p>2</p> Signup and view all the answers

    What is the purpose of using oil immersion lens during the cytological examination of the external ear canal?

    <p>To examine for bacteria and yeast</p> Signup and view all the answers

    [In the ear canal] Microscopically you should see none to occasional bacteria. Many of the bacteria are potentially pathogenic if and when the conditions are optimal to colonize in the ear canal. In bacterial infections the cytological examination can reveal large numbers of bacteria along with what cell?

    <p>Polymorphonuclear cells</p> Signup and view all the answers

    Normal flora of the ear canal can contain a small number of bacteria, which of the following is a normal inhabitant of the external ear canal and also the most common cause of mycotic otitis externa

    <p>Malassezia spp</p> Signup and view all the answers

    What is the characteristic shape of yeast cells observed during the cytological examination of the external ear canal?

    <p>Both A and C</p> Signup and view all the answers

    What is the recommended method for collecting a sample for otitis externa diagnosis?

    <p>Using a cotton swab to collect a sample from the ear canal</p> Signup and view all the answers

    What is the primary cause of mycotic otitis externa?

    <p>Malassezia spp</p> Signup and view all the answers

    What is the primary cause of otitis externa in dogs?

    <p>All of the above</p> Signup and view all the answers

    What is the characteristic of the squamous epithelial cells during Estrus?

    <p>Cells with no nucleus and abundant basophilic cytoplasm</p> Signup and view all the answers

    What is the primary cause of Clostridium perfringens infections in the intestines?

    <p>All of the above</p> Signup and view all the answers

    What is the longest stage of the canine estrous cycle?

    <p>Diestrus</p> Signup and view all the answers

    Malassezia (formerly known as Pityrosporum spp) is a

    <p>Budding, gram positive yeast</p> Signup and view all the answers

    Method for diagnosing Clostridium perfringens infections in the intestines?

    <p>All of the above</p> Signup and view all the answers

    What is the term for the false pregnancy seen in dogs?

    <p>Pseudocyesis</p> Signup and view all the answers

    What is the recommended stain to use during the fecal cytology examination?

    <p>Diff-Quik</p> Signup and view all the answers

    What is the main advantage of the non-aspiration technique in highly vascular areas?

    <p>It reduces the risk of blood contamination.</p> Signup and view all the answers

    What is the blood smear technique used for?

    <p>To prepare slides from samples with large fluid components</p> Signup and view all the answers

    This method takes some experience Inexperience hands the cytologic smear is often unreadable because too many of the cells are ruptured or is insufficiently spread

    <p>Squash preparation</p> Signup and view all the answers

    What are the benefits of the combination technique

    <p>All of the above</p> Signup and view all the answers

    What kind of areas will you use a nonaspiration technique on

    <p>Highly vascular areas</p> Signup and view all the answers

    What is the purpose of making multiple smears from an aspirate?

    <p>To increase the chances of getting a good quality slide.</p> Signup and view all the answers

    What is the disadvantage of using large needles for aspiration?

    <p>It increases the risk of blood contamination.</p> Signup and view all the answers

    What is the purpose of blotting the tissue before making a slide?

    <p>To prevent blood contamination during slide preparation.</p> Signup and view all the answers

    Why is it important that no downward pressure be applied when using the squash preparation method?

    <p>To prevent the rupture of fragile cells.</p> Signup and view all the answers

    What is the primary advantage of using scrapings to collect cytologic samples?

    <p>It yields a large number of cells, especially on firm lesions</p> Signup and view all the answers

    Why is it important to clean blood and tissue before making an imprint smear?

    <p>All of the above</p> Signup and view all the answers

    What is the main limitation of using impression smears to collect cytologic samples?

    <p>They often yield a small number of cells and are contaminated with bacteria and blood</p> Signup and view all the answers

    What is the primary difference between impression smears and scrapings?

    <p>The technique used to collect cells</p> Signup and view all the answers

    When is it recommended to use a combination of techniques to collect cytologic samples?

    <p>When the lesion is difficult to access</p> Signup and view all the answers

    What is the primary reason for leaving some smears unstained when collecting cytologic samples?

    <p>To allow for further testing or analysis</p> Signup and view all the answers

    Cocci is

    <p>Gram positive</p> Signup and view all the answers

    Why is it important to prepare the site before collecting cytologic samples?

    <p>To ensure the site is clean and free of debris</p> Signup and view all the answers

    What is the primary difference between blood smear technique and squash preparation?

    <p>The technique used to collect cells</p> Signup and view all the answers

    What is a disadvantage of using large needles for aspiration?

    <p>It increases the risk of blood contamination</p> Signup and view all the answers

    When unidentifiable yeast or hyphae are observed in the ear what should be done?

    <p>Culture</p> Signup and view all the answers

    What is the primary purpose of the Combination Technique?

    <p>To combine the advantages of the Blood Smear and Squash Techniques</p> Signup and view all the answers

    What is the primary advantage of the Starfish Technique?

    <p>It is a modification of the Squash Technique</p> Signup and view all the answers

    Otodectes cynotis is a common problem in dogs and cats characterized by

    <p>Reddish brown or black granular discharge is seen</p> Signup and view all the answers

    What is a common problem encountered when using the Squash Technique?

    <p>Inexperienced hands</p> Signup and view all the answers

    Clostridium perfringens is

    <p>Both A and C</p> Signup and view all the answers

    Certain factors may cause CP organisms to undergo enteric sporulation, during the sporulation process the enterotoxin is released, what does enterotoxin production cause

    <p>Causes intestinal damage and fluid accumulation in the intestines</p> Signup and view all the answers

    Preparation of collection site depends on site of collection. What kind of preparation does a superficial site require

    <p>Preparation similar to a venipuncture</p> Signup and view all the answers

    What is the primary characteristic of vaginal epithelial cells during Anestrus?

    <p>Large and cuboidal with large nuclei</p> Signup and view all the answers

    Preparation of collection site depends on the site of collection. What does microbiologic test require

    <p>Generally do not require a surgical prep (depends on sample needed)</p> Signup and view all the answers

    Smear preparation technique used depends on?

    <p>All of the above</p> Signup and view all the answers

    When should you leave some smears unstained

    <p>When sending to an outside lab</p> Signup and view all the answers

    Why should you imprint ulcers before cleaning

    <p>Highest bacterial count</p> Signup and view all the answers

    When imprinting cutaneous lesions or tissues collected during surgery/necropsy what must you do

    <p>Clean blood and tissue before imprinting</p> Signup and view all the answers

    What is the primary reason for leaving some smears unstained when collecting cytologic samples?

    <p>To send samples to an outside lab</p> Signup and view all the answers

    Always cytologic findings must be interpreted along with history and clinical signs The Four stages of estrus are:

    <p>Anestrus, proestrus, estrus, diestrus</p> Signup and view all the answers

    4 types of otitis externa

    <p>Acute, chronic, bilateral, unilateral</p> Signup and view all the answers

    Signs of otitis externa (7)

    <p>shaking head, mal odor, pain, pruritus, alopecia around the ear, otic discharge, excoriation</p> Signup and view all the answers

    Conformation of the ear canal • Long , narrow ear canal • Traps moisture, foreign debris, and glandular secretions • _______ ___ in the external ear canal will also inhibit ventilation and increase the retention of cerumen

    <p>excessive hair</p> Signup and view all the answers

    • Animal’s habits can lead to a predisposition for otitis For example

    <p>Hunting dogs, swimmers, allergies, parasites, bacteria, yeast, tumors, trauma</p> Signup and view all the answers

    • Obtain ear swag sample ____ otoscope examination has been performed

    <p>After</p> Signup and view all the answers

    [ear] Cytological examination should be evaluated for (7)

    <p>Bacteria, yeast, fungal hyphae, mites, neutrophils, cerumen, neoplastic cells</p> Signup and view all the answers

    Findings that must be recorded for bacteria (5)

    <p>type and morphology, grouping, quantity, FOV</p> Signup and view all the answers

    Findings that must be recorded for yeast (2)

    <p>Quantity and FOV</p> Signup and view all the answers

    Things that CP can cause (5)

    <p>Diarrhea, gas, abdominal discomfort, occassional hemorrhagic diarrhea, occasional vomiting</p> Signup and view all the answers

    Slide preparation for mites includes (4)

    <p>Mineral oil, swab with or without oil, slide, 10x observation</p> Signup and view all the answers

    Fecal smear steps (3) and 4 to identify?

    <p>make thin smear, heat fix or air dry, stain with diff quick, under oil to identify spores</p> Signup and view all the answers

    Smear preparation tips Cytologic samples can be collected in 4 ways, what are they

    <p>Imprints, scraping, swabbing, aspirating</p> Signup and view all the answers

    • Prepared from tissues collected during necropsy, surgery, or external lesions • Advantage o Collection yields large number of cells especially on firm lesions where it is difficult to obtain cells

    <p>imprints</p> Signup and view all the answers

    • Prepared from tissues collected during necropsy, surgery, or external lesions Disadvantage o Difficulty in collecting them o Superficial nature of the collection procedure often does not give a true representation of cells. This hinders their use as a diagnostic tool

    <p>Scrapings</p> Signup and view all the answers

    Method of collection generally used only when imprints, scrapings and aspirates are not possible because of location Such as fistulous tracts, vaginal cytology, ears

    <p>Swabs</p> Signup and view all the answers

    Things/areas that can be FNA (not specific organs) (4)

    <p>Lymph nodes, skin, internal organs, masses in thorax or abdomen</p> Signup and view all the answers

    Internal organs that can be FNA

    <p>liver, spleen, kidneys, lungs, prostate, thyroid</p> Signup and view all the answers

    Aspiration of the surrounding tissue should be avoided

    <p>Large mass = Redirect needle without releasing negative pressure, release pressure before withdrawing from mass Small mass = Relieve negative pressure during redirection, negative pressure is applied only when the needle is static Tissue biopsy = Sampling of piece of tissue for cytologic and/or histopathologic examination</p> <ul> <li>= -</li> </ul> Signup and view all the answers

    When performing an FNA the mass, tissue or organ is identified how

    <p>All of the above</p> Signup and view all the answers

    Why is FNA a preferred method for obtaining samples from masses

    <p>Both A and B</p> Signup and view all the answers

    The FNA method (aspiration or nonaspiration) depends on what

    <p>All of the above</p> Signup and view all the answers

    FNA technique, ___ gauge needle, __mL syringe, the softer the tissue the __ the needle and syringe (separate with commas)

    <p>25, 6, softer</p> Signup and view all the answers

    Larger needles often leads to poor quality slides, Tissue cores tend to be aspirated, which results in poor yield of free cells suitable for diagnosis, Tend to cause greater blood contamination

    <p>True</p> Signup and view all the answers

    Smaller needles often leads to poor quality slides, Tissue cores tend to be aspirated, which results in poor yield of free cells suitable for diagnosis, Tend to cause greater blood contamination

    <p>False</p> Signup and view all the answers

    Size of syringe is influenced by the consistency of the tissue being aspirated (12ml over all good average), 3ml good for lymph nodes, Fibrosarcoma requires a larger syringe because of firmness of the tumor

    <p>True</p> Signup and view all the answers

    Size of syringe is NOT influenced by the consistency of the tissue being aspirated (12ml over all good average), Any size is good for lymph nodes, Fibrosarcoma requires a smaller syringe because of firmness of the tumor

    <p>False</p> Signup and view all the answers

    At this point your needle is out of the mass o Remove needle from ___ of syringe o Draw ___ into the syringe o ___ needle onto the syringe o Depress the plunger ____ o Expelling aspirate onto the ___ of a slide o The needle should be ____ to the slide to allow for a single droplet of material vs a spray Separate with commas

    <p>hub, air, replace, rapidly, close</p> Signup and view all the answers

    • Modification technique of the original nonaspiration method o Using a ____ gauge needle on a 3 cc syringe o Draw a few cc of air into the syringe barrel ___ collection this allows for rapid expulsion of the material onto the slide o Grasp needle and syringe at the ___ for maximum control o Mass ___ with free hand o Needle is ____ o Move the needle back and forth in a _____ motion trying to stay on the same tract o This allows cells to be collected by ___ and tissue pressure o careful not to contaminate sample with ____ tissues o Withdraw the needle ____ releasing pressure o Quickly expel material onto slide o Smear promptly o Optimal to perform multiple attempt at various sites Separate with a comma

    <p>small, before, hub, stabilized, inserted, stabbing, cutting, surrounding, without</p> Signup and view all the answers

    A tissue biopsy is the sampling of a piece of tissue for cytologic and/or histopathologic examination. It consists of : Gentle ____ with a ____, Then excision, a ____ biopsy can be used or ______-guided biopsy . Fix tissue samples for histopathologic examination in 10% _____ _____-buffered formalin. To ensure adequate fixation, place slabs of tissue no more than __cm wide in fluid (10% formalin). Fill tight jars containing formalin at approximately 10 times the specimen's _______. With large tissues, remove the sample to a ____ jar with ____ formalin once fixed for 34 hours (separate with a comma)

    <p>abrasian, blade, punch, endoscope, neutral, phosphate, 1, volume, smaller, less</p> Signup and view all the answers

    With endoscope specimens, gently ____ specimens collected by endoscopy from the tip of the endoscope with ____ ____ . ____ the specimen gently on a paper towel to remove ____ . Then place on a small piece of _____ ______ ______ and allow the tissue to dry on the "". Once done, ______ or ____ specimens with the attached "___" specimen side down in fixative.(separate with commas)

    <p>flush, sterile, saline, blot, excess, blood, wooden, tongue, depressor, splint, immerse, float, splint</p> Signup and view all the answers

    Most common problems of tissue biopsies are ___ cells seen, lesions do not _____ well (mesenchymal neoplasms). Needle may ____ the site of lesion during collection. ____ sample collection. inadequate _______ ______ (aspiration technique). slow or ______ needle passes (nonaspiration technique) ... (separate with commas)

    <p>few, exfoliate, miss, timid, negative, pressure, shallow</p> Signup and view all the answers

    • Anytime you can visualize material in the hub of the needle you should _____ and ______ ______ ______ (separate with a comma)

    <p>stop, make, smears, immediately</p> Signup and view all the answers

    • Blood seen on impression smears often results from a ______ to _____ tissue before making slides (separate with a comma)

    <p>failure, blot</p> Signup and view all the answers

    In poorly prepared slides high cell yield is _______ when slides are prepared poorly. Smears should be made immediately to prevent ___ or _____. If a lot of material is recovered be prepared to make ___ slides or ___. Too thick of smears are useless as there are not enough _____. Smears should be air dried _____. Make ____ ____ good quality slides as possible to submit to ______. Cells are often fragile and break apart easily so having more than 1 or 2 slides can be beneficial

    <p>useless, drying, clotting, 2, more, cells, immediately, as, many, lab,</p> Signup and view all the answers

    For smears for solid masses, several methods can be used. The method used depends on 2 things. List them . (combination of methods is acceptable)

    <p>Experience of the person, characteristics of the sample</p> Signup and view all the answers

    Squash preparation method consists of: _____ aspirate onto the ______ of the slide. Place a ____ spreader slide over the aspirate ______. Quickly and smoothly ____ across the first slide. It is important that no ______ pressure be applied because this causes ______ of cells. (separate with commas)

    <p>Expel, middle, second, horizontally, slide, downward, rupture</p> Signup and view all the answers

    Steps of the combination technique: Place aspirate on ____ of the slide, draw back spreader slide until it contacts ____ of the sample and push the slide _____(blood smear). Next, place a spreader slide _______ over the back 1/3rd of the sample. Allow sample to ______. Quickly and smoothly _____ the slide (squash method). The ____ third of the saple is left untouched . (Separate with commas)

    <p>middle, 1/3rd, forward, horizontally, spread, slide, middle.</p> Signup and view all the answers

    One modification of the squash technique is the _____, where you drag the aspirate with the tip of a needle to make a ______

    <p>Starfish, starfish</p> Signup and view all the answers

    Line concentration techniquer is ________________________ (one sentence

    <p>Blood smear without the feathered edge</p> Signup and view all the answers

    Preparation of smears from fluids should be made immediately after collection, be sure to collect in ____. Smears can be prepared from ___ or _____ samples. _____ if low cellularity is expected. You may use the blood smear, squash prep, and line smear technique, _____, _____ and _____ influence selection. (separate with commas)

    <p>EDTA, fresh, centrifuged, centrifuge, cellularity, viscosity, homogeneity</p> Signup and view all the answers

    In centrifuged samples, excessive fluid keeps cells from spreading well. Be sure to centrifuge at a rate similar to ____ samples. Once done the supernatant is separated from the sediment, resuspend the sediment in a few drops of _____. ____ ______ or _______ ___ technique is used (separate with commas)

    <p>urine, supernatant, blood, smear, squash, prep</p> Signup and view all the answers

    _____ _____ is used for samples of low cellularity

    <p>Line smear</p> Signup and view all the answers

    What are the 3 stain types that offer excellent nuclear and cytoplasmic detail and also allow viewers to see through the layers of the cell to evaluate nuclear and nucleolar changes (separate each stain with a comma)

    <p>romanowsky type stain, new methylene blue, papanicolau stain</p> Signup and view all the answers

    Cocci shaped bacteria are usually

    <p>Both A and B</p> Signup and view all the answers

    Rod shaped bacteria are usually

    <p>Both A and B</p> Signup and view all the answers

    What is the characteristic feature of Clostridium perfringens spores observed during the fecal cytology examination?

    <p>Both A and C</p> Signup and view all the answers

    Imprints are usually performed on

    <p>Both A and B</p> Signup and view all the answers

    Exfoliative vaginal cytology is useful adjunct to the history and clinical examination in determining the stage of the estrous cycle in bitches and queens, but it also assists with?

    <p>Both A and C</p> Signup and view all the answers

    Clostridium perfringens is??????

    <p>Both A and C</p> Signup and view all the answers

    Disadvantages of the imprint method

    <p>Both A and C</p> Signup and view all the answers

    Study Notes

    Cytological Examination of the External Ear Canal

    • Evaluate for bacteria, yeast, fungal hyphae, mites, neutrophils, cerumen, and neoplastic cells
    • Record findings, including bacteria type and morphology, quantity, and field of view (FOV)

    Normal Ear Canal Flora

    • Small number of bacteria, typically cocci-shaped (Staphylococcus and Streptococcus spp.) or rod-shaped (Pseudomonas aeruginosa or Proteus mirabilis)
    • Presence of Malassezia spp. (formerly Pityrosporum spp.) yeast, a normal inhabitant of the external ear canal

    Mites

    • Common problem in dogs and cats, caused by Otodectes cynotis
    • Characterized by reddish-brown or black granular discharge, with secondary bacterial and/or yeast infections possible

    Fecal Cytology

    • Examine for Clostridium perfringens (CP) and other enteric bacteria
    • Identify spores, which are typically present in high numbers in cases of CP enterotoxin production

    Diagnosis of CP Enterotoxin Production

    • Must be examined during a clinical episode
    • Use fecal cytology or Enterotoxin ELISA test to diagnose
    • High numbers of CP spores on fecal cytology serve as a quick and simple screening test

    Vaginal Cytology

    • Evaluate for stages of estrus, including anestrus, proestrus, estrus, and diestrus
    • Identify cornified epithelial cells, neutrophils, and RBCs to determine stage of estrus cycle

    Stages of Estrus

    • Anestrus: no vulvar swelling, noncornified squamous epithelium, and few neutrophils
    • Proestrus: swollen vulva, reddish discharge, and noncornified cells becoming cornified
    • Estrus: discharge changes to pinkish-white, all squamous epithelial cells are cornified, and neutrophils present
    • Diestrus: vulvar swelling and discharge decrease, noncornified epithelial cells predominate, and neutrophils present

    Cytology Sample Collection and Preparation

    Preparation of Collection Site

    • Depends on the site of collection, e.g. superficial site requires preparation similar to a venipuncture, while surgical preparation is required for aspirates of internal organs, joints, body cavities, bone marrow, and spinal tap
    • Microbiologic tests generally do not require a surgical prep, depending on the sample needed

    Imprints

    • Used for external lesions, especially on living animals, but also on tissue removed during surgery or necropsy
    • Easy to collect, but results may not always be satisfactory
    • Collect fewer cells than other methods, and may be contaminated with bacteria, blood, or other substances

    Scrapings

    • Prepared from tissues collected during necropsy, surgery, or external lesions
    • Advantage: yields a large number of cells, especially on firm lesions where it is difficult to obtain cells
    • Disadvantage: difficulty in collecting them, and may not give a true representation of cells due to the superficial nature of the collection procedure

    Fine Needle Aspirates (FNA)

    • Used for lymph nodes, skin, subcutis, internal organs, and masses in the thorax or abdomen
    • Preferred method for obtaining samples from masses, as it avoids superficial contamination and allows for cells to be collected from various areas within the lesion
    • Collected either by aspiration or nonaspiration techniques, depending on the type of lesion and the preference and experience of the collector

    Aspiration Technique

    • Mass is held firmly to aid penetration and control of the needle
    • Needle with syringe attached is introduced into the center of the mass, and strong negative pressure is applied
    • Multiple areas of the mass should be sampled, and aspiration of the surrounding tissue should be avoided

    Nonaspiration Technique

    • Works well for highly vascular areas, and involves using a small gauge needle on a 3 cc syringe
    • Modification of the original nonaspiration method, where a few cc of air are drawn into the syringe barrel prior to collection
    • Allows for rapid expulsion of the material onto the slide, and cells are collected by cutting and tissue pressure

    Tissue Biopsy

    • Sampling of a piece of tissue for cytologic and/or histopathologic examination
    • Methods include gentle abrasion with a blade, excision, punch biopsy, and endoscope-guided biopsy
    • Fix tissue samples in 10% neutral phosphate-buffered formalin for histopathologic examination

    Endoscope Specimens

    • Gently flush specimens collected by endoscopy with sterile saline, and blot gently on a paper towel to remove excess blood
    • Immerse or float specimens with attached "splint" in fixative

    Common Problems

    • Too few cells seen, due to lesion not exfoliating well, needle missing the site of the lesion, timid sample collection, inadequate negative pressure, or slow or shallow needle passes
    • Blood contamination, which can be avoided by using nonaspiration technique or smaller needles
    • Poorly prepared slides, which can be improved by making smears immediately, using the correct technique, and submitting multiple slides to the lab

    Smear Preparation

    • There are several methods, including squash, blood smear, combination, and line smear techniques
    • Method used depends on the experience of the person, characteristics of the sample, and combination of methods
    • Smears should be made immediately after collection, and cells should be spread evenly to prevent drying or clotting

    Stains

    • Romanowsky type stain, New Methylene Blue, and Papanicolaou stain are commonly used
    • Papanicolaou stain provides excellent nuclear and cytoplasmic detail, but requires multiple steps and considerable time

    Cytology Sample Collection and Preparation

    Preparation of Collection Site

    • Depends on the site of collection, e.g. superficial site requires preparation similar to a venipuncture, while surgical preparation is required for aspirates of internal organs, joints, body cavities, bone marrow, and spinal tap
    • Microbiologic tests generally do not require a surgical prep, depending on the sample needed

    Imprints

    • Used for external lesions, especially on living animals, but also on tissue removed during surgery or necropsy
    • Easy to collect, but results may not always be satisfactory
    • Collect fewer cells than other methods, and may be contaminated with bacteria, blood, or other substances

    Scrapings

    • Prepared from tissues collected during necropsy, surgery, or external lesions
    • Advantage: yields a large number of cells, especially on firm lesions where it is difficult to obtain cells
    • Disadvantage: difficulty in collecting them, and may not give a true representation of cells due to the superficial nature of the collection procedure

    Fine Needle Aspirates (FNA)

    • Used for lymph nodes, skin, subcutis, internal organs, and masses in the thorax or abdomen
    • Preferred method for obtaining samples from masses, as it avoids superficial contamination and allows for cells to be collected from various areas within the lesion
    • Collected either by aspiration or nonaspiration techniques, depending on the type of lesion and the preference and experience of the collector

    Aspiration Technique

    • Mass is held firmly to aid penetration and control of the needle
    • Needle with syringe attached is introduced into the center of the mass, and strong negative pressure is applied
    • Multiple areas of the mass should be sampled, and aspiration of the surrounding tissue should be avoided

    Nonaspiration Technique

    • Works well for highly vascular areas, and involves using a small gauge needle on a 3 cc syringe
    • Modification of the original nonaspiration method, where a few cc of air are drawn into the syringe barrel prior to collection
    • Allows for rapid expulsion of the material onto the slide, and cells are collected by cutting and tissue pressure

    Tissue Biopsy

    • Sampling of a piece of tissue for cytologic and/or histopathologic examination
    • Methods include gentle abrasion with a blade, excision, punch biopsy, and endoscope-guided biopsy
    • Fix tissue samples in 10% neutral phosphate-buffered formalin for histopathologic examination

    Endoscope Specimens

    • Gently flush specimens collected by endoscopy with sterile saline, and blot gently on a paper towel to remove excess blood
    • Immerse or float specimens with attached "splint" in fixative

    Common Problems

    • Too few cells seen, due to lesion not exfoliating well, needle missing the site of the lesion, timid sample collection, inadequate negative pressure, or slow or shallow needle passes
    • Blood contamination, which can be avoided by using nonaspiration technique or smaller needles
    • Poorly prepared slides, which can be improved by making smears immediately, using the correct technique, and submitting multiple slides to the lab

    Smear Preparation

    • There are several methods, including squash, blood smear, combination, and line smear techniques
    • Method used depends on the experience of the person, characteristics of the sample, and combination of methods
    • Smears should be made immediately after collection, and cells should be spread evenly to prevent drying or clotting

    Stains

    • Romanowsky type stain, New Methylene Blue, and Papanicolaou stain are commonly used
    • Papanicolaou stain provides excellent nuclear and cytoplasmic detail, but requires multiple steps and considerable time

    Vaginal Cytology

    • Exfoliative vaginal cytology is useful in determining the stage of the estrous cycle in bitches and queens.
    • Assists with optimal timing of mating or artificial insemination in small animals.
    • Helps in diagnosing vaginitis.
    • Technique:
      • Animal is restrained in a standing position.
      • Vulva is cleaned and rinsed.
      • Lubricated speculum is introduced into the vulva.
      • Sterile premoistened (saline) swab is introduced into the vagina.
      • Swab is rotated and removed.
      • Swab is gently rolled on 2 or 3 clean microscope slides.
      • Slides are dried and stained with Diff-Quik.
    • Evaluation of smear:
      • Cytologic findings must be interpreted with history and clinical signs.
      • Four stages of estrus: anestrus, proestrus, estrus, and diestrus.
      • Rising levels of estrogen cause vaginal epithelium to become "cornified".
      • Tracking changes in morphology of desquamated vaginal epithelial cells provides a convenient means of assaying changes in estrogen levels.

    Cytological Examination of the Vaginal Smear

    • Evaluate for:
      • Bacteria (rods, cocci).
      • Yeast (Malassezia spp).
      • Fungal hyphae.
      • Mites.
      • Neutrophils.
      • Cerumen.
      • Neoplastic cells.
    • Recording findings:
      • Bacteria: type/morphology, grouping, quantity, and FOV.
      • Yeast: quantity and FOV.
      • Record anything else seen and quantify.

    Ear Cytology

    • Cytological examination of the external ear canal is useful in diagnosing otitis externa.
    • Common causes of otitis externa:
      • Bacteria.
      • Yeast (Malassezia spp).
      • Mites.
      • Fungal hyphae.
    • Collection of sample:
      • Cotton swab.
      • Obtain sample after otoscope examination has been performed.
      • Roll swab on a clean slide.
    • Evaluation of smear:
      • Bacteria: rods, cocci, and quantity.
      • Yeast: "peanut-shaped" organisms, dark blue staining, budding.
      • Mites: reddish-brown or black granular discharge, secondary bacteria and/or yeast.
      • Fungal hyphae: rare.

    Fecal Cytology

    • Cytological examination of feces is useful in diagnosing Clostridium perfringens (CP) infection.
    • CP: enteric anaerobic gram-positive rod, normal enteric bacteria, sporulation.
    • Enterotoxin production causes intestinal damage and fluid accumulation in the intestines.
    • Diagnosis:
      • Must be examined during a clinical episode.
      • Enterotoxin ELISA test.
      • Fecal cytology: high numbers of CP spores serves as a quick and simple screening test.
    • In-house test:
      • Fecal smear: make a thin smear of the feces, heat fix or air dry, stain with Diff-Quik.
      • Identify spores under oil immersion.

    Review

    • Technique and evaluation of exfoliative vaginal cytology.
    • Stages of estrus: clinical signs and cytological findings.
    • Common causes of otitis externa.
    • Cytological examination of the external ear canal.
    • Fecal cytology.

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    Description

    This quiz assesses your knowledge of the steps involved in examining an ear swab for various microorganisms, including bacteria, yeast, and parasites. It covers the procedures for collecting and staining the swab, as well as the evaluation of findings.

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