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Laboratory Sciences
Final Exam Review - From Preprinted Notes (Coagulation done)
| Question | Answer |
|---|---|
| Define Necropsy. | Exam of an animal after its death to determine any diseases that were present during life |
| Define Pathology. | The study of disease |
| Define Histopathology. | Microscopic evaluation of tissue disease |
| Define Lesion. | Any abnormality of tissue |
| Define Pathogenesis. | Sequence of events that leads to disease |
| What is the purpose of a necropsy? | Determine the disease process or processes that led to the animal’s death. Evaluate the accuracy of clinical diagnosis. Evaluate the effectiveness or consequences of therapeutic measures. Diagnostic for herd health; prevention of herd disease. |
| What six things do you need for an effective necropsy? | Knowledge of anatomy and gross pathology. Systematic evaluation of the body. Sample collection. Description of history, live animal exam, necropsy findings. Images of findings. Proper preparation and shipping of samples. |
| Necropsy What types of tests are we collecting samples for? | Histopathology Cytology Microbiology: Bacteria, Virus, Fungus Toxicology Parasitology |
| Necropsy What to remember when collecting samples? | Proper collection, preservation, & shipping of samples is essential accuracy! Culture samples should be collected rapidly after necropsy begins. Intestinal evaluation & samples: last. Contact lab for collection/preservation rules (Toxocology especially). |
| What are some biohazard risks when performing a necropsy? | Large amounts of body fluids and blood may be exposed during necropsy. Personnel should always consider biohazard concerns to themselves and lab personnel. (Rabies, Mycobacterium, Leptospirosis, Aspergillosis) Label all potential biohazards clearly! |
| Rabies | |
| Mycobacterium | |
| Leptospirosis | |
| Aspergillosis | |
| Necropsy Remember, in the event of a rabies suspect: | Head must be submitted to the state diagnostic lab within 24 hours of death with correct forms. It must be kept chilled/not frozen. Necropsy shouldn't be done until after rabies results. Techs shouldn't decapitate rabies suspects unless trained to. |
| Necropsy Toxicology | If poisoning is suspected: stomach contents, urine and blood serum should be collected and frozen. Kidney/liver samples should be collected (saved in formalin and by rapid freezing techniques) Any additional lesions samples of organ saved in formalin. |
| What to remember about body preservation for necropsy? | Should be performed soon after death. If delayed body should be refrigerated/rapidly cooled. After death tissue decay begins in delicate tissues (brain, spinal cord, lungs, eyes). If a body is frozen/thawed tissue damage results in poor histopathology. |
| Necropsy Equipment Protective clothing | Apron or disposable gown. Exam or rubber gloves. Eye wear. Surgical masks. Protective footwear (large animal). |
| Necropsy Equipment Surgical instruments | Dedicated to necropsy exams. Disposable: scalpels, scissors, forceps, bone cutters. |
| Necropsy Equipment Other equipment | Saws. Necropsy Knives. Sharpeners. |
| What to include in the necropsy report? | Signalment. History. Lesion Descriptions: location, number, color, size, shape, consistency, distribution, and odor. |
| Necropsy Procedure External Exam | Prior to dissection, the external body should be examined and findings recorded and images taken as appropriate. |
| Necropsy Procedure Internal Exam | Must be systematic so that no abnormalities are missed. (Thorax, Abdomen, Throat/mouth/nose, Neurologic System) |
| Necropsy Collection of Samples | Formalin 10% buffered: most common tissue fixative. Tissue specimens should be < 1cm thick for correct fixing. Ample formalin must be used (10x volum0.e of tissue). Formalin should be handled with gloves in well-ventilated area: irritant, carcinogen. |
| Necropsy Internal Exam | Midline incision of skin entire length. Limbs reflected by cutting attachments. Samples: joints, skin, lymph nodes, masses, testes, eyes, sciatic nerve, muscle. Abdominal wall muscles reflected. Ribs cut on costalchondral junction. Ribcage removed whole. |
| Necropsy Thorax | All organs are inspected, images taken. Samples of lung, lymph nodes, fluids taken. Heart and great vessels are dissected and samples taken. |
| Necropsy Abdomen | Organs inspected in situ. GI tract is cut at esophagus (in neck) and rectum. GI tract may be removed as one unit and evaluated last. Other organs are sampled: liver, kidneys, lymph nodes, reproductive structures, gall bladder, urinary bladder, spleen. |
| Necropsy Throat/mouth/nose | Midline incision in neck is made to inspect tissues of neck, mouth and nose. |
| Necropsy Neurologic Exam | Head removed. Skull cut to expose brain. After visual exam: brain is removed/sampled. Eval of spinal cord: all dorsal spinous processes removed. Cerebral spinal fluid collected (brain's ventricles largest sample). Eval of brain & spinal cord is skilled. |
| Necropsy Gastrointestinal Tract | Serosa is evaluated. Tract is opened and examined from cranial to caudal. All lesions samples. If no lesions are seen, samples of esophagus, stomach, duodenum, ileum, jejunum, and colon should be taken. |
| Cosmetic Necropsy | Limited procedure. Performed when owners wish body returned. Abdominal incision and diaphragm cut. Reach up into chest: trachea/esophagus cut to remove organs. Colon/urethra tied off. Abdomen filled, skin sewn, eyelids shut, animal brushed. |
| Necropsy Sample Submission: Things to remember. | Absolute diagnosis is rarely made at the time of necropsy. Results of histopathology, culture and toxicology evaluation are usually where diagnosis is made. Careful sample submission is vital! Remember, garbage in – garbage out! |
| Necropsy Sample Submission: packing. | Fill out appropriate forms. Place in ziploc. Package samples to protect against leakage. Label all biohazards. Protect cytology samples from formalin. Keep unfixed specimens chilled. Arrange for shipping/pick-up. Call laboratory to clarify procedure. |
| Evaluation of the Hematopoetic System What three things do you always need: | Patient history, Physical exam with lymph node palpation, Complete Blood Count (CBC). |
| Whole blood is made up two components. What are they. What will you find in each component? | Plasma: liquid component (water, protein, sugars, lipids, electrolytes). Cells: erythrocytes, leukocytes, thrombocytes |
| Cells of blood What is the main function of red cells? | carry oxygen |
| Cells of blood What is the main function of platelets? | initiate blood clotting |
| Cells of blood What is the main function of white cells? | Immune system: Granulocytes (Neutrophils, Basophils, Eosinophils) and Monocytes - direct attack. Lymphocytes - create antibodies |
| How large is a normal canine RBC? | 7 um |
| White Cells - Granulocytes Segmented Nucleus & Granules Neutrophil. What is their function? | Clear-grey granules. Phagocytize bacteria and other debris and then dissolve with enzymes. |
| White Cells - Granulocytes Segmented Nucleus & Granules Eosinophil. What is their function? | Pink/red granules. Ingest antigen-antibody complexes. Mediate allergic/parasitic responses. |
| White Cells - Granulocytes Segmented Nucleus & Granules Basophil. What is their function? | Blue to Purple granules. Mediate allergic/parasitic responses. |
| How large is a normal canine neutrophil (PMN)? | 14 um |
| White Cells - Agranulocytes - No granules Lymphocyte | Round nucleus. 7-9 um in diameter. Some originate in lymph nodes or thymus. T-cells: cell-mediated immunity–B-cells: humoral immunity (antibody production)–NK cells: cause apoptosis of cells infected with virus. |
| White Cells - Agranulocytes - No granules Monocyte | Variable nucleus. Large @ 12 - 14 um. |
| White Cells - Agranulocytes - No granules Lymphocyte: Function of T-cells | cell-mediated immunity (conTrols immune response) |
| White Cells - Agranulocytes - No granules Lymphocyte: Function of B-cells | humoral immunity (antiBody production) |
| White Cells - Agranulocytes - No granules Lymphocyte: Function of Natural Killer cells (NK) | Cause apoptosis (seppuku) of cells infected with virus. |
| White Cells - Agranulocytes - No granules What is the function of the Monocyte? | Remove cellular debris; chronic inflammatory process. Process antigen and present to lymphocytes. |
| Where is the thymus? | At the thoracic inlet. It shrinks as the animal ages. |
| Describe red cells: | No nucleus in mammals. Disc shaped (biconcave). |
| Describe platelets: | No nucleus. Clear to light blue. Cell fragments of cytoplasm. |
| What is the blue-purple area in a platelet? | DNA |
| The CBC (complete blood count) includes what two basic evaluations? | Counts: exact numbers of red cells, white cells and platelets. Morphology: evaluation of these cells. |
| The CBC includes five additional evaluations of blood. Name them: | Plasma protein (TP) in g/dl. Hemoglobin concentration in g/dl. PCV or HCT in %. Erythrocyte indices (MCV, MCH, MCHC). Reticulocyte count (if patient is anemic) |
| Blood collection. Name two important things to remember: | Fast flowing blood is best. Sample needs to be collected without clotting. |
| What three factors can cause clotting when performing a blood draw? | Tissue trauma. Prolonged draw time. Not inverting tube appropriately/wrong ratio of blood to coagulant. |
| Why is fast flowing blood best when drawing blood? | It prevents platelets "clumps." Clumped platelets can not be counted. Jugular blood samples are best but rapid Cephalic may be acceptable. |
| Vacutainers | Collection tubes that contain various substances to prepare blood for specific tests. Tubes contain a vacuum so that they self-fill the correct amount of blood if tops not removed. Sterile inside tube. |
| What is the preferred collection tube for CBC samples? | EDTA tube (lavender top) |
| What two factors should you keep in mind for blood smears for CBCs? | Feathered edge required. Fresh from the patient is best. |
| Ideal angle to hold second slide for doing a blood smear? | 30 degrees (also the ideal angle for IV blood collection) |
| Hematopoetic System Packed Cell Volume (PCV) | % of spun blood that is red cells |
| Hematopoetic System Buffy Coat | Layer of white cells in spun blood |
| Hematopoetic System Total Protein | Measurement of solids dissolved in plasma. Determined by Refractometer. |
| Hematopoetic System - Plasma Color Red | Hemolysis: due to rupture of red cells |
| Hematopoetic System - Plasma Color Cloudy -> White | Lipemia: due to lipids |
| Hematopoetic System - Plasma Color Yellow | Icteric: due to excess bilirubin |
| How do you perform a PCV and read it properly? | Blood is placed in hematocrit tube & sealed with clay. The sample is spun in hematocrit centrifuge for 3-5 minutes. Remember counterbalance. The spun sample is aligned on reader card and the top of the red cell layer is read. |
| How do you use a Refractometer? | After the PCV has been determined, using at least 2 tubes, and the result has been recorded, gently sample one tube over the serum region. Dot a small amount onto the refractometer careful to leave no glass behind. |
| What four things are important to remember when using a Refractometer? | Place cover over the sample. Hold instrument up to the light. Read result and record. Immediately clean refractometer with stream of distilled water and dry. |
| Determining CBC Red cell count is helpful in: | Evaluation for anemia or polycythemia (excess RBCs) |
| Determining CBC Red cell Morphology is indicates: | Size of red cells indicates low iron or increased production. Appearance of red cells may indicate specific disease processes. |
| Determining CBC - Total White blood count (WBC) High (Leukocytosis) | Certain infections, Inflammation, Certain neoplasia (cancer), Hyperadrenocorticism or corticosteroid use, Stress (esp. cats) |
| Determining CBC - Total White blood count (WBC) Low (Leukopenia) | Certain infections, Bone marrow damage, Hypoadrenocorticism |
| Determining CBC - Platelet count High (Thrombocytosis) | Hyperadrenocorticism / steroid use, Rare bone marrow neoplasia |
| Determining CBC - Platelet count Low (Thrombocytopenia) | Massive hemorrhage, Certain infections (usually tick-borne), Immune system destruction, Risk for clotting disorder |
| Determining CBC Automated Analyzers Facts: | Reliability varies greatly. Professional labs analyzer results need direct evaluation verification (by hand). Machine counts cells by size. Abnormal sized cells are counted wrong. No machine is capable of: morphology interpretation/ID of blood parasites. |
| What is one way to perform a manual count. What equipment can you use? | Hemocytometer: counting chamber to determine number of cells per ul. Set up as a grid pattern to count number of either WBC/RBC per ul. The cells are counted systematically from left to right & averaged. |
| WBC Differential Count | Knowing the exact count of each white cell type provides important body state information. A blood smear is stained with a Wright’s stain (Trichrome). 100 cells in the monolayer are counted and ID'd. The % of each is multiplied by total WBC. |
| Knowing the exact count of each white cell type provides much information on body state. What is this called: | The Differential Count. |
| Hematology When first evaluating a blood smear what is an important first step? | Beginning with low power: scanning to locate the best area of the smear for examination at higher magnification. This is a good time to evaluate the smear for validity as well: are their platelet clumps? Scanning for parasites is also a good idea. |
| WBC Differential & Absolute Count Example: | .78 neutrophils x 9500 total WBC = 7410 PMN .12 lymphocytes x 9500 = 1140 lymph .08 eosinophils x 9500 = 760 eos .02 monocytes x 9500 = 190 mono 0 basophils= 0 baso |
| Why is Absolute Count better? | Two animals have the same differential–70% neutrophils (PMNs). But the absolute neutrophil counts are 10,000 and 1,000. Which one is obviously at risk for dying? |
| Erythrocyte Indices List one pro and one con: | Assist in classification of anemias. Provide little pertinent information if the animal is not anemic. |
| Erythrocyte Indices Name the three: | Mean Corpuscular Volume (MCV). Mean Corpuscular Hemoglobin (MCH). Mean Corpuscular Hemoglobin Concentration (MCHC). |
| Where on the slide do we perform the differential? | Monolayer |
| What three parameters do we examine to ID WBC? | Size of cell (compared to RBC), shape of nucleus, and cytoplasm contents. |
| How do you perform Platelet Estimates? | Count # of platelets per oil field. We average the count from several fields. Place this in a formula to generate a platelet estimate. Normal is 200,000-250,000 platelets per ul. |
| What six parameters do we use to evaluate erythrocyte morphology? | Arrangement. Color. Size. Shape. Number. Unique features – (nuclei, inclusions, parasites). |
| Erythrocyte Morphology Number: | Anemia: decreased RBC number. Polycythemia: increased RBC number. |
| Erythrocyte Morphology Color: | Hypochromasia: decreased blue staining due to decreased iron content. Polychromasia: Multiple colors in population of RBCs due to the presence of young bluer cells. |
| Erythrocyte Morphology Size: | Macrocytosis – enlarged due to young red cells–Microcytosis – small due to decreased Hgb content–Anisocytosis – variable size in population |
| Erythrocyte Morphology Arrangement: | Rouleaux: “stacking of red cells” dispersed with saline. Agglutination: clumping of red cells due to antibody attachment. |
| Erythrocyte Morphology Shape (Poikilocytes) | Spherocytes, Schistocytes, Echinocytes. Others: Acanthocytes, Dacryocytes, Codocytes, Keratocytes, Eccentrocytes |
| Erythrocyte Morphology | |
| Hypochromasia | As a descriptor of erythroid cells as seen on a blood film, it refers to the appearance of increased central pallor with a thin rim of cytoplasm. Usually due to low iron anemia. |
| Polychromasia | Usually associated with regenerative anemia, young RBCs are bluer due to RNA. |
| Poikilocytes | Erythrocytes with abnormal shape for the species at hand. Some have fairly specific diagnostic significance, while other forms are very non-specific. |
| Spherocyte | Red cells which have assumed the form of a sphere rather than the normal discoid shape. Show loss of central pallor. Often due to spleen removing damaged areas. |
| Schistocyte | Red cell fragments. Generally taken to reflect mechanical injury to erythrocytes. Tumor of spleen or liver are common causes. |
| Nucleated RBCs | Bone marrow injury. Lead poisoning. Acute myeloid leukemia and myelodysplasia. Abnormal splenic function. Physiologic: Miniature Schnauzers, Poodles and Dachshunds can have small numbers of nRBCs in health. Regenerative response to anemia. |
| Reticulocytes | Slightly immature, anucleate red cells that contain RNA. The RNA, which binds basic dyes, may be sufficient to impart a blue-gray tint, called polychromatophilia. A new Methylene Blue is special stain to help ID them. |
| Erythrocyte Morphology - Artifacts Crenated red cells: also known as echinocytes or burr cells | Red cells have changed from a disc shape to spheres covered with short, sharply pointed projections. Slow Drying of the smear and aging of blood in the tube are the most common causes of artifactual crenation |
| Erythrocyte Morphology - Artifacts “Water” artifact | Water contamination of stain or slide. Causes “bubble” appearance. |
| Platelets | First defense against hemorrhage. Produced by megakaryocytes in bone marrow. Thrombocytopenia= too few platelets. Thrombocytosis=increased numbers of platelets |
| Platelet Estimate | Determine the average number of platelets in 5 to 10 oil immersion fields. At this magnification each platelet is approximately equal to 15,000 platelets/μl. Not admissable if clumps are present on slide or clots in tube. |
| What is a normal Platelet Estimate in dogs? Cats? | Normal platelet counts in dogs/cats 10-30/oil field. 200,000 – 500,000/ul |
| Name and describe three abnormal finding involving Leukocytes. | Leukopenia: too few white blood cells. Neutropenia: too few neutrophils. Leukocytosis: an increase in the number of circulating white blood cells, often due to infection or inflammation. |
| What are Band Cells? | Immature Granulocyte. Nucleus is curved to “U” shaped. Once the “waist” of the nucleus pinches down 1/3 or more compared to nucleus ends, then classified as segmented. |
| What is a Toxic Neutrophil? | These changes are morphological abnormalities acquired during maturation under conditions that intensely stimulate their production and shorten maturation time in marrow. Less condensed chromatin and bluer cytoplasm due to retention of ribosomal RNA. |
| What is a “Reactive" lymphocytes? | Larger cells with coarse (mature) chromatin, and deep blue cytoplasm. |
| Lymphoblast | Immature lymphocyte: large, dark nucleus with rim of cytoplasm. Cell is large. Due to Inflammation or Leukemia. |
| Cystocentesis Collection Method | Palpate bladder. Must be sufficiently distended to isolate it. Should be performed on calm easily restrained patients. Use 20 or 22 gauge needle, 1 to 1&1/2 in long & sterile 10-2 mL syringe. RBC’s can be evident due to iatrogenic bladder trauma. |
| Cystocentesis Collection Method Needle placement | Female dogs and all cats: insert needle ventral midline, caudal to umbilicus. (alcohol puddle trick). Male dogs: insert caudal to umbilicus and to the side of the sheath. |
| Describe three important points to remember when performing a cystocentesis. | Never redirect needle once in abdomen-trauma to bladder/other organs. Never put same needle into abdomen twice-hitting colon /contaminating abdomen. Always release negative pressure on syringe plunger before withdrawing needle-possible urine in abdomen. |
| Describe catheterization method. | External genitalia should be cleansed. Always use sterile urinary catheters and gloves. A small amount of sterile lubricant is placed on the tip of the catheter. The distal end of the catheter is designed to attach a sterile syringe. |
| Key points to remember when performing catheterization method. | Females: a speculum may be used to help visualize the urethral orifice. Cats are almost always anesthetized for this procedure. Care must be taken to avoid trauma to the sensitive urethral mucosa. |
| Voided or “free catch” samples | Collect in a clean not necessarily sterile container. Wash the vulva/prepuce area. A midstream sample is less likely to be contaminated. For cats, a litter box is used containing non-absorbent granules. |
| When collecting free catch urine, what situation necessitates collecting initial stream instead of midstream? | When dogs cease urinating when collection attempts are made. |
| Manual Bladder Expression | With the animal standing or in lateral recumbency, the bladder is palpated in caudal abdomen and gentle, steady pressure is applied. Do not exert such pressure as to injure or rupture the bladder. (In general not a recommended sample due to trauma risk). |
| In what instances should manual bladder expression be avoided? | On animals whose urethra may be obstructed or whose bladder wall may be fragile. |
| Urine - Storage & Handling Ideally, samples should be analyzed within __ _____ to __ ____ to avoid post-collection artifacts and degenerative changes. | 30 minutes, 1 hour |
| If immediate analysis is not possible, what preserves most urine constituents for an additional 6 to 12 hours? | refrigeration |
| If a urine sample is refrigerated, what must be done with it before it is analyzed? | brought to room temperature |
| Which urinary test is most important to complete before refrigerating urine? | SG - Specific Gravity, Artifact crystals may form as urine cools |
| What six things initially evaluated and recorded during urinalysis? | Color, volume, how was it obtained, odor, turbidity/clarity/transparency, specific gravity |
| Urine is normally? (color) | Light yellow to amber |
| Colorless urine will have? | low SG |
| Dark to brown urine, generally has? | high SG |
| Yellow-brown to green urine may indicate? | That it is bile containing |
| Red to red-brown urine indicates? What do you call this? | RBC's in urine. Hematuria. |
| What may alter the color of urine, other than disease process? | Drug. May make urine: red, green, or blue |
| Why is volume important to evaluate? | May be important to assess kidney function or endocrine disease. Certain parameters are evaluated with volume in mind. To estimate urine output, a 24-hour volume should be determined. |
| Odor of urine is not always diagnostically important, but it may indicate? | Normal varies among species/gender. Samples left standing at room temp may develop an odor of ammonia due to bacterial growth. A sweet or fruity odor indicates ketones, most commonly found with diabetes mellitus. |
| Transparency of urine is noted as: | clear, slightly cloudy, cloudy, or turbid (flocculent) |
| Urinalysis: clear samples can | go directly to chemistry analysis. Generally do not yield much sediment upon centrifugation. |
| Urinalysis: cloudy samples are | often centrifuged before the next step in urinalysis: chemical properties. |
| True or false: Reagent strips will not be as accurate a read as the refractometer. | True |
| Specific gravity is? | The weight (density)compared to an equal amount of distilled water. |
| Three things to remember about SpGr: | May be determined before/after centrifugation. The particles that settle will have little or no effect on reading. It can be affected by time of collection, eating & drinking habits. Yields info on hydration & ability of kidneys to concentrate or dilute. |
| What are two common tests performed on a refractometer? | Specific gravity and Blood Serum Protein |
| Three points to remember about refractometer care: | Careful to not scratch plate when applying sample. Rinse with distilled water & wipe dry with lens paper after each use. (do not allow samples to dry on refractometer) |
| What is the specific gravity (concentration) of plasma (osmolality)? | 1.010-1.012 |
| If isosthenuric (urine sp gr is 1.010-1.012) and patient dehydrated, this indicates? | Organ failure: kidney, liver, endocrine |
| What is a normal range of SG for canine urine? | 1.018-1.030 |
| What is a normal range of SG for feline urine? | 1.020-1.035 |
| Reagent strip chemistries are performed on _____ urine, unless the urine is turbid (cloudy). | unspun |
| True or false: each square on a reagent strip has a specific wait time. | True |
| What two methods can be used to apply urine to reagent strips? | Apply drop by drop to each square with a syringe/syringe & needle. Dip it in the urine sample and turn on side to allow excess to drip off. |
| Reagent strips should be stored at _____ temperature, and container is to remain ______. Do not use _____ strips. They provide inaccurate results. | room, closed, expired |
| Microscopic Exam of Urine Step 1 | CENTRIFUGE sample (10 mL) for 5 minutes. |
| Microscopic Exam of Urine Step 2 | Pour off the ‘supernate’ (liquid above sediment). |
| Microscopic Exam of Urine Step 3 | Sediment remains, although normal urine of domestics does not a large amount. |
| Microscopic Exam of Urine Step 4 | Pipette one drop of sediment on microscope slide, cover with coverslip. |
| Microscopic Exam of Urine Step 5 | A second drop can be placed on the same slide and stained with ‘sedi-stain’, cover with coverslip. |
| Microscopic Findings Epithelial Cells - What three types are usually found? | Squamous, Transitional, Renal |
| Microscopic Findings Epithelial Cells - SQUAMOUS | Large angular borders small nuclei. They originate from lining of distal urethra and vagina or prepuce. Not generally indicative of disease. |
| Microscopic Findings Epithelial Cells - TRANSITIONAL | Medium-sized and oval spindled. Found lining the proximal urethra, bladder, ureters, renal pelvis. May occur in groups. |
| Microscopic Findings Epithelial Cells - RENAL | Small round cells that may indicate tubular degeneration. |
| Urine stain will enhance identification of: | Cells in sample: RBC, WBC, Epithelial cells/casts) |
| Stains may add artifacts of: | crystals, bacteria, stain debris |
| It is recommended that you evaluate unstained sample for crystals and bacteria; then add drop of stain to: | evaluate for cellular components |
| RBC appearance can be affected by three things: | urine concentration, pH, time |
| In unstained fresh sediment they appear: | Colorless or yellowish, small, round, slightly refractile. |
| True or false: RBCs may be confused with fat droplets, but do not float in and out of focus like fat droplets. | True |
| In concentrated urine, RBC’s _____, _____ (ruffled edges), slightly darker and granular. | shrink, crenate |
| What are ghost cells? | RBC that have imbibed water in dilute urine |
| In urine, leukocytes appear: | Round, granular, larger than erythrocytes, smaller than epithelial cells. Leukocytes are spherical, dull gray or greenish-yellow. |
| Presence of more that ___-___ per high power field indicates inflammation of urinary or urogenital tract, dependent upon collection method. | 5-8 |
| What is Pyuria? What does it indicate? | Excessive WBC’s in urine. Inflammatory or infectious process. |
| What are casts in urine? | Prominent in urine. They are elongated structures composed of a matrix of protein from plasma and mucoprotein secreted by the renal tubules. |
| Casts _____ in _____ urine, so it is important to access examining fresh urine, since urine becomes alkaline from standing. | dissolve, alkaline |
| A large number of casts indicates: | A lesion in the renal tubules, but the number of casts is not an indicator of severity of disease. |
| True or False: Casts and cellular debris originate high up in the urinary tract. Usually suggestive of injury to kidneys from inflammation or lack of blood flow. | True |
| What are five main types of urine casts? | hyaline, cellular, granular, waxy, fatty |
| TYPES OF URINARY CASTS Hyaline | Colorless and semitransparent. May be difficult to see w/o reducing microscope light. They indicate mild renal glomerular (bundle of capillaries) leakage. |
| TYPES OF URINARY CASTS Cellular | Recognizable cells they may be epithelial, erythrocyte (indicating renal hemorrhage), or leukocyte (renal inflammation). |
| TYPES OF URINARY CASTS Granular | Derived from degenerating cells or cellular casts. Most common. Seen in large numbers they indicate more severe kidney damage. |
| TYPES OF URINARY CASTS Waxy | Wide and homogenous (uniform), usually with distinct blunt or squared ends. Indicate a more chronic tubular lesion. |
| TYPES OF URINARY CASTS Fatty | Contain fat droplets that appear as refractile bodies. Often seen in cats with renal disease, and occasionally in dogs with diabetes mellitus. Large numbers indicate degeneration of renal tubules. |
| True or False: Conditions that lead to crystal formation may form urinary calculi (stones) but crystals are not good predictors of stones. | True |
| The type of crystal formation depends upon four things. Name them. | pH, concentration, temperature, and solubility of elements. |
| If a technician does these two things, it is likely to increase the numbers of crystals present in urine: | refrigerate sample, allow the urine sample to stand and then cool |
| Crystals are generally recorded as: | occasional, moderate, many or +1 - +4. |
| Calcium Oxalate Dihydrate crystals are _____ in urine. What do they look like? | Common. Envelope-like, with an X across the crystal |
| Calcium Oxalate – Monohydrate (coffin shaped) are found in the urine under what condition? | Ethylene Glycol Poisoning (antifreeze) |
| Struvite: (triple phosphate crystals or ammonium, magnesium phosphate) are found in: | Slightly acidic urine. |
| Triple Phosphate Crystals - Struvite are prism shaped. What two treatments can be successful at getting rid of them? | Diet, Treatment of infections present |
| Ammonium Biurate crystals are seen in slightly acidic, neurtal, or alkaline urine. They are brown in color, round, with long irregular spicules "thorn apple" shape. They are most common in: | Animals with liver disease |
| Calcium Carbonate crystals | Commonly seen in horse and rabbit urine. They are of no clinical significance. They often have a “dumbell” shape. (slit cell or half an apple) |
| Amorphous Phosphate crystals | MEANS HAVING NO DEFINITE SHAPE OR FORM. COMMON IN ALKALINE URINE AND APPEAR AS A GRANULAR PRECIPITATE. |
| What three microorganisms can be found in urine? | bacteria, fungi, protozoa |
| Normal aseptically captured urine is ___ ___ of bacteria. | free of |
| BACTERIA CAN ONLY BE IDENTIFIED MICROSCOPICALLY. They may be | ROUND (COCCI) OR ROD SHAPED (BACILLI), USUALLY REFRACT LIGHT AND APPEAR TO BE QUIVERING DUE TO BROWNIAN MOVEMENT (oscillating movement of microscopic particles). |
| True or false: Some parasites of the urinary tract include Capillaria plica, a bladder worm of dogs and cats, and Dioctophyma renale, a kidney worm of dogs. | True |
| Uroliths are: | Calculi (stones) composed of various minerals found anywhere in the urinary tract. |
| Why are uroliths diagnostically important? | They may cause blockage of urine outflow from bladder into urethra; lodge in urethra causing severe, acute inability to urinate; or remain in bladder causing inflammation and bleeding. |
| Visual exam ___ ___ accurate for urolith identity. | is not |
| The CBC - Complete Blood Count Counts.... | exact numbers of red cells, white cells, and platelets. |
| The CBC - Complete Blood Count Morphology.... | evaluation of these cells: Normal vs Abnormal appearance |
| The CBC - Complete Blood Count Other than Counts & Morphology it also includes: | Plasma protein (TP) in g/dl, Hemoglobin concentration in g/dl, PCV or HCT %, Erythrocyte indices: MCV, MCH, MCHC, Reticulocyte count (if patient is anemic) |
| The CBC - Complete Blood Count Plasma protein (TP) is reported as: | g/dl |
| The CBC - Complete Blood Count Hemoglobin concentration is reported as: | g/dl |
| The CBC - Complete Blood Count Plasma protein (TP) | Total protein is the sum concentration of all individual serum proteins (g/dL). Usually evaluated using a Refractometer. |
| The CBC - Complete Blood Count Hemoglobin concentration | Used to check for anemia |
| The CBC - Complete Blood Count PCV or HCT is reported as: | a percent % |
| The CBC - Complete Blood Count Erythrocyte indices. Name the three: | MCV, MCH, MCHC |
| The CBC - Complete Blood Count Erythrocyte indices - MCV | Mean Corpuscular Volume. Blood test measures the average size of red blood cells. |
| The CBC - Complete Blood Count Erythrocyte indices - MCH | Mean Corpuscular Hemoglobin. The average amount of Hemoglobin protein in each red blood cell. It carries oxygen around in the body. |
| The CBC - Complete Blood Count Erythrocyte indices - MCHC | Mean Corpuscular Hemoglobin Concentration. A measure of the average concentration of hemoglobin inside a single red blood cell, but takes into account how big or small the red blood cells are. |
| The CBC - Complete Blood Count When is a Reticulocyte count performed? | If patient anemic |
| Functions of Granulocytes Neutrophils | Phagocytize bacteria and other debris and then dissolve with enzymes. |
| Functions of Granulocytes Eosinophil | Ingest antigen-antibody complexes. Mediate allergic/parasitic responses. |
| Ingest antigen-antibody complexes Functions of Granulocytes Basophils | Mediate allergic/parasitic responses. |
| Function of Agranulocytes Lymphocytes | T-cells: cell-mediated immunity. B-cells: humoral immunity (antibody production). NK cells: cause apoptosis of cells infected with virus. |
| Function of Agranulocytes Monocytes | Remove cellular debris; chronic inflammatory process Process antigen and present to lymphocytes. |
| Platelet Estimates How do you perform this count? | First evaluate slide on low power for clumps. If clumps are present, estimate can’t be performed! If not, count in 10 fields on oil objective lens. Choose area of the monolayer where RBCs are “crowded." Average this number and then multiply by 20,000. |
| Platelet Estimates How are the results recorded? | µL (microliter) Example: >200,000/µL |
| Feline Red Cell Characteristics: | Smaller and more variable in shape than dog RBC. Slight to no central pallor. PCV is lower in cat 30-45 vs. 37-55 dog. Rouleaux “stacking” is normal for feline blood (not dog). |
| Name a reason that Heinz Bodies would be found in cat blood. | Common in cat blood damaged by oxidative change, such as onion/garlic toxicity. Confirmed with New Methylene Blue Stain. |
| Mycoplasma haemofelis (Hemobartonellosis) | Most cats are probably carriers of this blood parasite. Acquired from eating fleas. Asymptomatic unless concurrent disease, such as FeLV. Not stain precipitate! Small dark dots on and around RBC. |
| Feline Platelets Characteristics: | Large platelets are normal for cat. Platelet clumping is common in cats, makes platelet counts challenging. |
| Eosinophils in the ___ have round granules and a light blue cytoplasm. Eosinophils of the ___ have small rod-shaped orange granules that fill the cytoplasm. | dog, cat |
| _____ basophils have pale lavender granules and a long and folded ribbon-like nucleus. _____ basophils are packed with similar small, slightly oval pale lavender granules. | Canine, Feline |
| Evaluation of the Hematopoeitic System always needs these three things: | History, Physical exam with lymph node palpation, Complete Blood Count (CBC) |
| Hematopoeitic System Whole Blood - Plasma | Liquid component Water, proteins, sugars, lipids, electrolytes |
| Hematopoeitic System Whole Blood - Cells | Erythrocytes, Leukocytes, Thrombocytes |
| Hematology What is the first step when evaluating the slide? | Begin with low power scanning to locate the best area of the smear for examination at higher magnification. |
| Erythrocyte | Hemoglobin – O2 bearing molecule. Comprised of 4 sub units: Globin (binds to 1 O2 molecule), Heme (iron). Variation of morphology occurs between species. Nucleated RBCs in birds & reptiles. |
| Erythrocyte Morphology What five physical aspects of RBC to we need to evaluate? | Arrangement, Color, Size, Shape, Number, Unique features: nuclei, inclusions, parasites. |
| Erythrocyte Morphology Number - Anemia | Decreased RBC number |
| Erythrocyte Morphology Color - Hypochromasia | Decreased staining due to decreased iron content. As a descriptor of erythroid cells as seen on a blood film, it refers to the appearance of increased central pallor with a thin rim of cytoplasm. Usually due to low iron anemia. |
| Erythrocyte Morphology | |
| Erythrocyte Morphology Number - Polycythemia | Increased RBC number |
| Erythrocyte Morphology Color - Polychromasia | Multiple colors in population of RBCs due to the presence of young bluer cells. Usually associated with regenerative anemia, young RBCs are bluer due to RNA. |
| Erythrocyte Morphology Size - Macrocytosis | Enlarged due to young red cells |
| Erythrocyte Morphology Size - Microcytosis | Small due to decreased Hgb content |
| Erythrocyte Morphology Size - Anisocytosis | Variable size in population |
| Erythrocyte Morphology Arrangement - Rouleaux | “Stacking of red cells” dispersed with saline. |
| Erythrocyte Morphology Arrangement - Agglutination | Clumping of red cells due to antibody attachment. |
| Erythrocyte Morphology Shape - Spherocytes | Are red cells which have assumed the form of a sphere rather than the normal discoid shape. Show loss of central pallor. Often due to spleen removing damaged areas. |
| Erythrocyte Morphology Shape - Schistocytes | Red cell fragments. Generally taken to reflect mechanical injury to erythrocytes. Tumor of spleen or liver are common causes. |
| Erythrocyte Morphology Shape - Echinocytes - Artifacts | Crenated red cells also known burr cells. Have changed from a disc shape to spheres covered with short, sharply pointed projections. Slow drying of the smear and aging of blood in the tube are the most common causes of artifactual crenation. |
| Erythrocyte Morphology Poikilocytosis | Erythrocytes with abnormal shape for the species at hand. Some have fairly specific diagnostic significance, while other forms are very non-specific. |
| Erythrocyte Morphology Nucleated RBCs - Etiology | Bone marrow injury, lead poisoning, acute myeloid leukemia & myelodysplasia, abnormal splenic function. Physiologic: Miniature Schnauzers, Poodles, & Daschunds can have small numbers of nRBCs in health. Can also be a regenerative response to anemia. |
| Erythrocyte Morphology Reticulocytes | Slightly immature anucleate red cells that contain RNA which binds basic dyes may be sufficient to impart a blue-gray tint called polychromatophilia. New Methylene Blue is special stain used to ID them. |
| Erythrocyte Morphology Artifacts - Name and describe two. | “Water” artifact. Water contamination of stain or slide Causes “bubble” appearance & Stain Precipitate |
| Thrombocytes Platelets are the first defense against _____, and they are produced by _____ in the bone marrow. | hemorrhage, megakaryocytes |
| Platelets Thrombocytopenia | Too few platelets. |
| Platelets Thrombocytosis | Increased numbers of platelets. |
| Platelet Estimate | To estimate platelet number, determine the average number of platelets in 5 to 10 oil immersion fields. At this magnification, each platelet is approximately equal to 20,000 platelets/µl. |
| True or false: Platelets can not be counted or estimated if clumps are present on slide or clots in tube (report as “clumped”). | True |
| Platelet Estimate Normal platelet counts in dogs/cats 10-30/oil field | 200,000 – 500,000/ul |
| Abnormal Leukocytes Leukopenia | Too few white blood cells |
| Abnormal Leukocytes Neutropenia | Too few neutrophils. |
| Abnormal Leukocytes Leukocytosis | An increase in the number of circulating white blood cells, often due to infection or inflammation. |
| Band Cell | Immature Granulocyte. Nucleus is curved to “U” shaped. Once the “waist” of the nucleus pinches down 1/3 or more compared to nucleus ends, then it is classified as segmented. |
| Toxic Neutrophil Why & What does it look like? | Are morphologic abnormalities acquired during maturation under conditions that intensely stimulate production and shorten the maturation time in marrow. Less condensed chromatin and bluer cytoplasm due to retention of ribosomal RNA. |
| Toxic Neutrophil Most common cause? | Severe bacterial infection |
| Reactive Lymphocytes | Are larger cells with coarse (mature) chromatin, and deep blue cytoplasm. |
| Lymphoblast or “blast” | Immature lymphocyte – large, dark nucleus with rim of cytoplasm. Cell is large. Commonly caused by inflammation or Leukemia. |
| What does Leukemia mean? | A cancer of the blood or bone marrow. Bone marrow produces blood cells. It can develop due to a problem with blood cell production. It usually affects the leukocytes, or white blood cells. |
| Evaluated by hematology? | Cells |
| Contains blood proteins. Evaluated by clotting times and clinical chemistries? | Plasma |
| Plasma minus the clotting proteins. Evaluated by clinical chemistries? | Serum |
| Chemical evaluation can be performed on what normally occurring body fluid (name five) | Blood (serum and plasma), urine, cerebrospinal fluid (CSF), Synovial Fluid, Cavity Fluids (abdomen/chest: peritoneal or pleural) |
| Chemistries may include identifying and measuring amounts of what six components of blood? | Electrolytes, Metabolites, Gases, Protein/Enzymes, Antibodies, Hormones |
| Clinical chemistry is a piece of the puzzle. What four other diagnostics aid in the most accurate disease diagnosis? | History, Physical Exam/Observation, Radiographs/Ultrasounds, Cytology |
| A Piece of the Puzzle Clinical Chemistry: ALT, CK, K+ | Cellular Damage |
| A Piece of the Puzzle Clinical Chemistry: BUN, Na+, Cr | Hydration Status |
| A Piece of the Puzzle Clinical Chemistry: Cr, Bile Acids, pO2 | Organ Function |
| A Piece of the Puzzle True or False: Clinical Chemistry can be used to evaluate: exposure to specific infectious diseases and hormone production. | True |
| What can we do to help generate accurate results in clinical chemistry? | Fill out submission forms completely. Collect, store, ship sample appropriately. |
| What four things can interfere with accurate clinical chemistry results? | Drugs, Hemolysis, Lipemia, Icterus |
| Give two examples of two ways that drugs can interfere with accurate clinical chemistry results? | Corticosteriods (i.e. prednisone) and Phenobarbital will cause drug induction of liver enzymes. Sulfa antibiotics may cause crystals in urine. |
| Clinical Chemistry Test Tubes Red top: Contents and what is evaluated? | Crystal Activator. Serum. |
| Clinical Chemistry Test Tubes Tiger Top: Contents and what is evaluated? | Clot Activator Gel. Serum. |
| Clinical Chemistry Test Tubes Green Top: Contents and what is evaluated? | Lithium Heparin. Plasma. |
| Clinical Chemistry Test Tubes Gray Top: Contents and what is evaluated? | Sodium Fluoride. Glucose. |
| Clinical Chemistry Test Tubes Lavender Top: Contents and what is evaluated? | EDTA. Whole Blood Hematology. |
| Clinical Chemistry Test Tubes Light Blue Top: Contents and what is evaluated? | Sodium Citrate. Coagulation Times. |
| Clinical Chemistry Test Tubes Clear Top: Contents and what is evaluated? | Nothing within. Transfer tube; specimens to culture. |
| Ischemia (poor perfusion/blood circulation), Toxin, Trauma, Infection will result in cell ATP/Membrane damage. What are some possible chemistry indications? | Enzymatic release (CPK, ALT, Amylase, Lipase) into the blood stream or blood pH changes due to the release of K+, Lactate, or H+ into the blood. |
| Clinical Chemistry - How it Works Metabolic Trigger results in: | Increased production which is measured in the serum. |
| Clinical Chemistry - How it Works Metabolic Damage results in: | Decreased removal which is measured in the serum. |
| Clinical Chemistry - Exocrine Pancreas What is its function? | To produce digestive enzymes such as amylase and lipase. |
| Clinical Chemistry - Exocrine Pancreas Elevations of amylase and lipase suggest pancreas disease because: | A damaged pancreas leaks enzymes. |
| Clinical Chemistry - Exocrine Pancreas Low TLI (trypsin-like immunoreactivity) consistent with exocrine pancreatic insufficiency (EPI) is due to: | Decreased production of pancreatic enzymes. |
| Clinical Chemistry - Lungs pCO2 and pO2 can be measured from: | Blood. Arterial samples are the most accurate. |
| Clinical Chemistry - Lungs What is measured? | Gas exchange in the lungs. |
| Clinical Chemistry - Lungs What does it evaluate? | Lung compensation for metabolic diseases. |
| Clinical Chemistry - Liver What is its function? | To remove toxins from the body and the bile system assists in fat metabolism. |
| Clinical Chemistry - Liver What enzyme leaks from damaged liver cells the most in carnivores? | ALT - Alanine Aminotransferase |
| Clinical Chemistry - Liver Name two liver enzymes that can increase due to problems with other organs or medications. | AST - Aspartate Aminotransferase, AP - Alkaline phosphatase |
| Clinical Chemistry - Liver What is elevated with liver disease and red cell destruction? | Bilirubin |
| Clinical Chemistry - Liver What is one excellent test for liver function? | Bile acid study |
| Clinical Chemistry - Liver What protein made in the liver will show decrease in levels with severe intestinal or kidney disease? | Albumin |
| Clinical Chemistry - Kidneys What are their function? How do they accomplish this goal? | To maintain homeostasis. By removing toxins, maintaining water and electrolyte balance, and maintaining blood pressure and red count. |
| Clinical Chemistry - Urine How is concentration measured? What is the SG of water and normal urine? | Measured in specific gravity. SG of water is 1.000. Normal urine SG is 1.030. |
| Clinical Chemistry - Blood Chemistry - Kidney What test reflects the glomerular filtration rate of kidneys? | Blood Urea Nitrogen (BUN) |
| What is the elevation of BUN called? | azotemia |
| What is a more accurate test of kidney function, but is late to rise? | Creatinine |
| Kidney Disease - Secondary Problems Anemia (PCV) due to? | Decreased erythropoietin. |
| Kidney Disease - Secondary Problems Low Albumin due to? | Leaking nephron. |
| Kidney Disease - Secondary Problems Low blood pH due to? | Increased H+. |
| Endocrine Organs What are Parathyroids responsible for? | Regulation of calcium metabolism. (Parathormone, PTH) |
| Endocrine Organs What is the Thyroid responsible for? | Regulation of metabolic rate. (Thyroxine, T4) |
| Endocrine Organs Pancreas (Endocrine) is responsible for? | Glucose regulation. Insulin – lowers blood glucose. If no insulin = Diabetes Mellitus. Glucagon – raises blood glucose. |
| Endocrine Organs Adrenal glands are responsible for? | Hormones for stress response: Corticosteroids (prednisone-like). Excess corticosteroid = Cushing’s disease. Electrolyte balance: Mineralcorticoids. |
| Endocrine Organs Where is the Pituitary Gland located? What does it do? | Located in brain. It secretes hormones to communicate with other endocrine organs: Antidiuretic hormone, Gonadotropins, Growth Hormone. |
| Total Body Water is made up of what two divisions? | Intracellular Fluid (ICF) and Extracellular Fluid (ECF) which is found in the blood, intestinal tract, and cavity spaces. |
| True or false: The kidneys attempt to maintain balance between cations (positively charged electrolytes) and anions (negatively charged electrolytes). | True. Electrolytes: Anion (Cl- and HCO3-) and Cations (K+, Na+, H+) |
| pH is hydrogen ion concentration. Decrease in pH of blood is called ___. Increase in pH of blood is called ___. | acidosis, alkalosis |
| How do the kidneys and lungs attempt to maintain blood pH? | Constant adjustment of CO2, HCO3-, and H+. |
| Clinical Chemistry The liver is primarily evaluated by: | ALT (Alanine Aminotransferace), AP (Alkaline phosphatase), and Bilirubin. |
| Clinical Chemistry The exocrine pancreas is evaluated by: | Amylase & Lipase |
| Clinical Chemistry The endocrine pancreas is evaluated by: | Glucose and Insulin |
| Clinical Chemistry The adrenal cortex excretes: | Corticosteroids & Aldosterone |
| Clinical Chemistry The kidneys are evaluated by: | BUN & Creatinine. |
| What does centesis mean? | The act of puncturing a body cavity or organ with a hollow needle in order to draw out fluid. puncture - the act of puncturing or perforating. Abdominocentesis, paracentesis - centesis of the belly to remove fluid for diagnosis. |
| Cytology Terms Cytology | study of cells as individuals |
| Cytology Terms Histology | study of tissue |
| Cytology Terms aspirate | collecting cells via a suction technique |
| Cytology Terms biopsy | sample of tissue |
| Cytology Terms centesis | collecting fluid via a needle technique |
| Cytology Terms exudate | discharge of abnormal cause |
| Cytology Terms purulent | containing many neutrophils "pus" |
| Cytology Terms neoplasia | tumor (benign or malignant) |
| Cytology Terms malignant | cancer |
| Cytology Terms benign | not cancer |
| Cytology Terms Hyperplasia | excessive growth of normal tissue (benign) |
| Cytology Terms | |
| Cytology What is the primary purpose of this diagnostic technique? | Differentiate benign vs malignant. Other purposes include determining nature of inflammatory reactions: infectious, allergic, toxic. Identify causative agents when possible. |
| Cytology What are the advantages of this diagnostic technique? | Rapid sample collection & evaluation. Special equipment usually not needed. Sedation/anesthesia rarely needed. Several sample preparation options. Several sample staining options. Inexpensive. |
| Cytology What are the disadvantages of this diagnostic technique? | Doesn't provide the same information as histology which is more accurate. Histology provides tissue architecture info. Only a very limited number of cells being evaluated compared to entire mass/organ. Some organs/tumors/diseases yield little/no info. |
| Cytology Name six sample collection techniques. | Swabs. Scrapings. Imprints. Fine needle aspirate. Capillary technique. Centesis. |
| Cytology Why are swabs commonly used? For what type of samples: name four. | When aspirates or scrapings are not possible. Utilize a moist (saline is best), sterile cotton or rayon swab. Usages: Otic cytology, Nasal exudates, vaginal cytology, conjunctival cytology |
| Cytology Scrapings: Why and how? | Smears of scrapings from biopsy (tissue) samples or skin lesions. Scapel blade is held perpendicular to tissue and gently dragged across surface in one direction. Material that collects on blade is then dragged across glass slide. |
| Cytology Scrapings: What should we add to the blade if mites are suspected? | mineral oil |
| Cytology Imprints: how and what is one important thing that need to be done before the impression is taken? | Slide (or tape) is pressed against open wound or lesion. Slide is pressed against biopsy sample after excess blood/fluid removed with gauze. |
| Cytology Fine Needle Aspirates: cells are collected from? Give three examples. | Masses, lymph nodes, internal organs |
| Cytology Fine Needle Aspirates: If sample will be cultured or needle is entering a normally sterile site (joint, thorax, organ" then ___ ___ is used. List the three steps involved. | aseptic technique. Shave hair -> Surgical scrub then alcohol (3x) -> use only sterile items after prep. |
| Cytology Fine Needle Aspirates: Name three pieces of equipment necessary. | Syringe 6 or 12cc. Needle - 22g. Slides. 6 mL is generally used for more liquid samples. 12cc is for more thick or solid aspirates. (more negative pressure) |
| Cytology Fine Needle Aspirates: Name one very important action to remember when performing this procedure. | Negative pressure released prior to removing needle from tissue. |
| Cytology Capillary Technique: why is it generally chosen? | For delicate cells or samples |
| Cytology Capillary Technique: Describe in three steps. | Needle placed in tissue or lymph node without syringe. Needle is rapidly thrust back and forth with slight changes in direction. Needle attached to air filled syringe to blow sample onto slide. |
| Cytology Centesis: What is it? Why do we do it? | Needle insertion into any body organ or cavity to collect fluid. Diagnostic & Therapeutic. |
| Cytology Centesis: What are four. (named after site of insertion) | Cystocentesis – urinary bladder collection. Abdominocentesis – peritoneal cavity. Thoracocentesis – pleural cavity. Arthrocentesis - joint. |
| Cytology Centesis: True or false? Usually requires aseptic technique. | True |
| Cytology Centesis: Therapeutic procedures may require removal of ___ quantities of fluids. | large (100s of mL) |
| Cytology Centesis: What three items of information need to be recorded after procedure? | Amount of fluid from each site. Time of collection. Appearance of fluid- it may change from beginning to end. |
| Cytology Centesis: Samples are usually submitted for what three types of testing? | Culture: in culture media or red top. Cytology: slides or purple top tube. Possible chemical analysis (red top or grey top). |
| Cytology - Sample preparation techniques Fluid samples greater than ___ mL can be placed in ___ tube and refrigerated. | .25 mL, EDTA (purple top) |
| Cytology - Sample preparation techniques Most samples are placed directly ___ ___ ___ for evaluation. | onto glass slides |
| Cytology - Sample preparation techniques Similar to blood samples: the specimens will need to be ___ ___ the slide to maximize evaluation of individual cells. | spread across |
| Cytology - Sample preparation techniques Name five different techniques. | Smear prep. Squash prep. Starfish prep. Line smear. Concentration techniques. |
| Cytology - Sample preparation techniques Smear Prep - How to: | Spray sample in middle of clean slide. Lay spreader slide at 45 degree angle and back into 1/3 of sample. Pull spreader slide forward rapidly and smoothly. |
| Cytology - Sample preparation techniques Squash Prep - How to: | Place sample in middle of clean slide. Place spreader slide horizontal to sample slide. Spreader slide is pulled rapidly and smoothly across sample. At 45, 90, or 180 degree angle to sample slide. |
| Cytology - Sample preparation techniques Starfish Prep - Why & How? | Least damage to fragile cells but may lead to overly thick sample. Place sample in middle of clean slide. Drag sample with tip of needle in multiple directions. |
| Cytology - Sample preparation techniques Concentration Techniques: Why & How? | Some samples have very few cells in a large amount of fluid and concentrating the cells together in a small amount of liquid improves evaluation. Low speed centrifugation, Line smear, Cytocentrifugation. |
| Cytology - Sample preparation techniques Concentration Techniques: Low Speed Centrifugation | Fluid spun 5 minutes at 1000-1500 rpm. Supernatant (liquid) is poured off of sample button. Supernatant may need Total Protein eval. Remaining sample sediment is resuspended in drop of supernatant or plasma. Sample is placed on slide for smear prep. |
| Cytology - Sample preparation techniques Concentration Techniques: Line Prep | Place a drop of sample on clean slide. Begin to spread as for smear technique. Stop and lift spreader slide up when ¾ across slide, giving a line of sample across slide. Remember: Do not want feathered edge. |
| Cytology Fixing and Staining | Rapidly air dry or very gently heat dry slide. Fix slide with 95% methanol. Should be fresh and not contaminated. Fix for 2-5 minutes. |
| Cytology Fixing and Staining: Name two types. | Romanowsky Stains: Diff-Quick, Wright’s, Tri Chrome. New Methylene Blue. |
| Cytology Fixing and Staining: Romanowsky: Name three and why they are used. | Diff-Quick, Wright’s, Tri Chrome: Good cytoplasm and organism staining. Good general evaluation. Quick, easy, inexpensive staining method. |
| Cytology Fixing and Staining: New Methylene Blue: Why is it used? | Superior nuclear detail (Reticulocytes). Does not stain cytoplasm or RBCs. |
| Cytology Shipping Samples - Three things to remember. | Samples slides should be labeled and in a slide container after completely dried. Do not place unfixed slides in bag with formalin specimen. All forms should be filled out completely, including patient history. |
| Cytology Name one major indication for cytology. What are we differentiating? | To evaluate masses. Inflammation vs Neoplasia. |
| Cytology If inflammation, what are three causes? | Infection. Allergies. Irritants or Toxins. |
| Cytology If Neoplasia, what are two outcomes? | Malignant (cancer). Benign. |
| Cytology Evaluating Slides: List two steps. | Begin at low power to find area where individual cells can be evaluated. Evaluate clusters of tissue cells at higher power. |
| Cytology Evaluating Slides: What does inflammation look like? | Neutrophils present. Macrophages present. Chronic inflammation. Organisms present. Organisms inside of WBC. Eosinophils with allergic responses. |
| Cytology Evaluating Slides: Inflammation - neutrophils in non-toxic environment | hypersegmented |
| Cytology Evaluating Slides: Inflammation - Neutrophil Necrosis | Cell Death: Loss of cytoplasm membrane, Vacuoles, Blue-grey cytoplasm. Toxic environment. Bacteria present? |
| Cytology Evaluating Slides: Malignant Tumors - Four characteristics. | Alien cells for tissue (lung in liver). Fragile cells. High Nucleus:Cytoplasm. Large, abnormal Nuclei. Mitotic figures. Extreme variation in size. Variation in shape. Multiple nucleoli. |
| Cytology Evaluating Slides: Tissue Types. Name three basic. | Round cells. Epithelial cells. Mesenchymal cells. |
| Cytology Evaluating Slides: Tissue Types: Round cells. | Discrete, individual round cells. (i.e. blood cells, lymphocytes.) |
| Cytology Evaluating Slides: Tissue Types: Epithelial cells. | Occur in clusters, round to oval in shape. (i.e. glandular tissue, skin). |
| Cytology Evaluating Slides: Tissue Types: Mesenchymal cells. | Spindle cells. (i.e. connective tissue: muscle/bone) |
| Cytology Evaluating Slides: Tumor Types - Epithelial cells (Malignant) | Carcinoma |
| Cytology Evaluating Slides: Tumor Types - Mesenchymal cells (Malignant) | Sarcoma |
| Cytology Evaluating Slides: Normal Lymph Node | 90% small lymphocytes. Small lymphocytes are about the size or RBC. |
| Cytology Evaluating Slides: Tumor Types - Reactive Lymph Node | Neutrophils. Plasma cells. Lymphocytes making antibodies. Eccentric nucleus. |
| Cytology Evaluating Slides: Tumor Types - Lymphoma (neoplasia) | 75% or more large lymphocytes or blasts. |
| Cytology Evaluating Slides: Tumor Types - Mast Cell Tumor | Round cell tumor. Contains characteristic histamine granules. |
| Otic Cytology How is it done? | Sample collected on cotton swab prior to use of cleaning solutions or medications. Mineral oil on swab will help collection of mites. |
| Otic Cytology Why is a sample evaluation not a substitution for otic exam: Name three otic complications in need of eval. | Foreign bodies. Tumors. Ruptured tympanic membrane. |
| Otic Cytology What five things can we evaluate for? | Mites, Bacteria, Yeast, White cells, Tumor cells. |
| Otic Cytology Always remember! Tell your clients, too! | Sample for culture should be collected prior to any cleaning or medication. |
| Otic Cytology Most commonly diagnosis yeast? | Malassezzia Otitis |
| Achieving Hemostasis Name two: | Blood must be fluid. Blood must also coagulate (clot) at appropriate times: Rapid, Localized, Reversible. |
| Thrombosis is? | Inappropriate coagulation |
| Coagulation What are three major systems involved? | Vessel wall: Endothelium, Basement membrane. Platelets. Coagulation cascade: coagulation factors (proteins). |
| Antithrombogenic | favors fluid blood |
| Thrombogenic | favors clotting |
| Antithrombogenic Properties of Endothelium Describe three anti-platelet properties | Covers highly thrombogenic basement membrane. Uninjured endothelium does not bind platelets. PGI2 (prostacyclin) and NO from uninjured endothelium inhibit platelet binding. |
| Antithrombogenic Properties of Endothelium Nitric oxide (NO) | Also called nitrogen monoxide, is a colorless toxic gas that is formed by the oxidation of nitrogen. It performs important chemical signaling functions in the body and has various applications in medicine. |
| Antithrombogenic Properties of Endothelium Anticoagulant Properties | Heparin-like molecules: activate anti-thrombin III (inactivates active proteases.) |
| Antithrombogenic Properties of Endothelium protease | an enzyme which breaks down proteins and peptides. |
| Prothrombotic Properties of the Endothelium (when injured) Name three: | Synthesis of von Willebrand factor. Release of tissue factor. Activation of clotting factors via Ca bridges. |
| Coagulation - Mechanical Phase Describe two steps. | Vasoconstriction of vessel. Platelets are attracted to damaged endothelium of blood vessel: platelets attach to the wall and to each other and release thromboxane. |
| Coagulation - Chemical Phase What is it? | It is called the Coagulation Cascade: Intrinsic Pathway, Extrinsic Pathway, Common Pathway, Fibrin. Triggers for coagulation also initiate fibrinolysis. |
| Role of Vitamin K True or False: Some clotting factors require a modification before they are active in clotting. | True. These factors are II, VII, IX, X, and proteins C and S. This modification to active form requires Vitamin K. |
| Inhibitors of Coagulation Name three | Vitamin K antagonists (in vivo only). Calcium chelators (in vitro only): EDTA, Citrate, Oxalate. Heparin (in vivo and in vitro). |
| In vivo | (of a process) performed or taking place in a living organism. |
| In vitro | (of a process) performed or taking place in a test tube, culture dish, or elsewhere outside a living organism. |
| Fibrinolysis | Fibrin -> Plasmin -> Fibrin Degradation Products (FDP) |
| Coagulation 3 Major Systems Involved | Vessel wall: endothelium, basement membrane. Platelets. Coagulation cascade: coagulation factors (proteins). |
| Common Coagulopathies What are two forms? | Congenital & Acquired |
| Common Coagulopathies Congenital | Hemophilia & Von Willebrands |
| Common Coagulopathies Acquired | Rodenticide, Thrombocytopenia, Liver failure, DIC, Snake Bite |
| Common Coagulopathies What is Disseminated Intravascular Coagulation (DIC)? | A condition in which blood clots form throughout the body, blocking small blood vessels. Symptoms may include chest pain, shortness of breath, leg pain, problems speaking, or problems moving parts of the body. |
| Common Coagulopathies Petechiae | Small red or purple spot caused by bleeding into the skin. |
| Common Coagulopathies Bleeding Disorders | Platelets: Numbers too low for adequate protection of the body <50,000/ul (microliter). Platelets present, but not functioning properly. |
| Platelet Testing Platelet count | Accurate counts are not possible if platelets are clumped or clot is present in purple top tube. |
| Platelet Testing Platelet Function | Buccal Mucosal Bleeding Time (BMBT), Von Willebrand's factor (vWb factor) |
| Platelet Estimate - From Smear What is it? | Examine the area of the smear close to the feathered edge where the RBCs are not touching one another to determine the approximate number of platelets per field. |
| Platelet Estimate - From Smear Step 1 | Use the oil immersion lens to estimate the number of platelets per field. Look at 5-6 fields and take an average. 10 fields is ideal. A normal blood smear should have between 8 and 20 platelets per field in this area of the smear. |
| Platelet Estimate - From Smear Step 2 | Multiply the average by 15,000. A normal platelet estimate is between 200,000-399,999/ul (microliter). |
| Buccal Mucosal Bleeding Time Supplies: Name six possibilities | Stopwatch/timer, muzzle gauze, filter paper/gauze sponges. Bleeding time template device: Surgicutt or Simplate. Anesthetic or tranquilizer. Don't use narcotic drugs. Acceptable drugs: xylazine, propofol, barbiturates, inhalant gas anesthesia. |
| Buccal Mucosal Bleeding Time Procedure | Not painful but dogs must remain quiet and in position (up to 10-12 minutes). Sedation may be required for adequate restraint. Simplate device is triggered parallel to the lip margin. Blood flowing from wounds is gently blotted below incisions. |
| Buccal Mucosal Bleeding Time Afterward & What is the normal range? | Topical tissue adhesive (cyanoacrylate) can be applied to prevent re-bleeding. Two to four minutes. |
| Tests for Coagulation Factors Activated Clotting Time (ACT) | Evaluates all clotting factors except VII |
| Tests for Coagulation Factors Prothrombin Time (PT) | Evaluates Extrinsic & Common Pathways. Very sensitive to Vitamin K antagonists. |
| Tests for Coagulation Factors Partial Thromboplastin (PTT) | Evaluates Intrinsic pathway. Sensitive to Vitamin K antagonists and Hemophilia |
| Tests for Coagulation Factors - ACT Technique | Whole blood sample in diatomaceous earth. Invert to mix and place in warming block (98.6F) for one minute. Then check by inversion every 5-10 seconds until solid clot forms. Normal: 60-120 sec (dog), Normal 60-70 sec (cat). |
| Tests for Coagulation Factors PT and PTT - Four important notes | Test evaluated by analyzer. Collected in citrate tube (light blue). Very important that: tube is 2/3 full and that blood is from atraumatic venipuncture of jugular. |
| Tests for Coagulation Factors Name five other possible tests: | Von Willebrand (vWb), Fibrinogen, D-Dimer, Fibrin Degradation Products (FDP), PIVKA (proteins induced by vitamin k absence). |
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