1. SETTING UP A PRACTICE LABORATORY. General Clinical Pathology LECTURE 3. EQUIPMENT FOR VETERINARY PRACTICE LABORATORIES OVERVIEW

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1 Undergraduate Course in Veterinary Clinical Pathology Socrates - Erasmus programme General Clinical Pathology LECTURE 3. EQUIPMENT FOR VETERINARY PRACTICE LABORATORIES 3-1 OVERVIEW 1. Setting up a practice laboratory 2. Basic equipment 3. Mid-range equipment 4. Advanced equipment SETTING UP A PRACTICE LABORATORY 3-3 1

2 DIFFERENT TYPES OF EQUIPMENT BASIC MID-RANGE ADVANCED Expense Inexpensive Moderately expensive Very expensive Level of technical Little/none Moderate High expertise Level of Low Moderate High throughput 3-4 QUESTIONS TO ASK BEFORE SETTING UP A LABORATORY 1. What test results will I need urgently? 2. Which of these urgent tests can I perform in-house? 3. Is there a good veterinary laboratory nearby? 4. What non-urgent analyses may I want to run myself (or with the help of technical staff)? 5. Am I proficient enough in laboratory work to perform analyses myself (or supervise others in doing so)? 3-5 QUESTIONS TO ASK BEFORE SETTING UP A LABORATORY: ANSWERS 1,2. Answers almost identical whatever the practice EVERY PRACTICE SHOULD HAVE BASIC ITEMS OF EQUIPMENT 3,4,5. Great differences according to the kind of practice, practitioner, location, etc. MOST PRACTICES SHOULD HAVE MID-RANGE ITEMS OF EQUIPMENT SOME PRACTICES MAY CONSIDER ADVANCED ITEMS OF EQUIPMENT 3-6 2

3 OVERALL Equipment suitable for each practice will be different based on many factors such as: - throughput /size of practice - species seen - technical and clinical pathology expertise available - income of patients owners - vicinity to a referral laboratory BASIC EQUIPMENT 3-8 BASIC EQUIPMENT All veterinary clinics should have at least equipment suitable for specimen preparation specimen storage specimen shipment to specialised laboratories performance of urgent analyses safe working conditions safe disposal of biological material, glass, needles etc 3-9 3

4 BASIC EQUIPMENT Such equipment: - Provides quick results - Does not need specialised technical skills - Is inexpensive to purchase and operate 3-10 BASIC EQUIPMENT All veterinary clinics should have at least equipment suitable for specimen preparation specimen storage specimen shipment to specialised laboratories performance of urgent analyses safe working conditions safe disposal of biological material, glass, needles etc 3-11 BASIC EQUIPMENT : SPECIMEN PREPARATION - Centrifuges (bench & microhaematocrit) - Blood tubes with and without anticoagulants - Sterile tubes for culture and urinalysis - Pipettes - Slides, coverslips, stains

5 CENTRIFUGES Used with: - Blood to obtain plasma or serum - Blood to measure PCV - Urine to obtain sediment - Body fluids to concentrate cells Centrifuge with blood/urine and PCV rotors Bench centrifuge 3-13 BLOOD TUBES : colour codes are not standardised red dry (often silicone-coated) yellow green light blue purple grey lemon dry + gel Li or Na heparin Citrate EDTA fluoride ACD (citric acid, glucose) 3-14 STERILE TUBES FOR CULTURE AND URINALYSIS: EXAMPLE

6 PIPETTES - Volumetric pipettes to measure a precise volume of liquid - Dispensing pipettes semi-quantitative, to prepare & transfer specimens 3-16 SLIDES, COVER SLIPS, STAINS Stain system Slide staining rack 3-17 BASIC EQUIPMENT All veterinary clinics should have at least equipment suitable for specimen preparation specimen storage specimen shipment to specialised laboratories performance of urgent analyses safe working conditions safe disposal of biological material, glass, needles etc

7 BASIC EQUIPMENT: STORAGE & SHIPPING - Tubes, boxes, racks to store specimens - Identification system - Refrigerator, freezer and materials for keeping specimens cool in transit - Appropriate packaging and knowledge of postal regulations 3-19 STORAGE TUBES Plain non-anticoagulated tubes Eppendorf tubes Racks 3-20 BASIC EQUIPMENT All veterinary clinics should have at least equipment suitable for specimen preparation specimen storage specimen shipment to specialised laboratories performance of urgent analyses safe working conditions safe disposal of biological material, glass, needles etc

8 BASIC EQUIPMENT: URGENT/BASIC ANALYSES Equipment for quick, simple tests at minimun cost; providing basic and important clinical information. Includes: - Microscope - Refractometer - Tests or test kits for clinical biochemistry analysis (biochemical strips and urine dipsticks) - Simple tests for coagulation defects - Simple serological tests (snap tests) 3-22 MICROSCOPE Used in combination with: - slides / stains - haemocytometer and pipettes It allows the clinician to analyze: - RBC, WBC and platelet count, blood smears - urine sediment - cytological samples - rumen fluid - parasitology tests 3-23 REFRACTOMETER (CLINICAL) Measures the refractive index of a solution : n D (= for water) - in plasma/serum: total protein concentration - in urine: specific gravity - in effusion fluids: protein concentration (semi-quantitative) - not suitable for measuring: protein in urine and CSF, osmolality of plasma/serum - STANDARDISE PERIODICALLY BEFORE USE, IE CHECK SG = WITH DISTILLED WATER - FOR ACCURATE RESULTS, SERUM/PLASMA OR URINE MUST NOT BE HAEMOLYSED OR TURBID

9 VETERINARY REFRACTOMETER AND ITS SCALE 3-25 STRIPS FOR BIOCHEMISTRY ANALYSIS: glucose, urea, ketones They are used with a drop of whole blood, but in some cases may be used also with serum/plasma or urine GLUCOMETERS - Very useful to help control of blood glucose levels in animals with diabetes mellitus - Need only 1 drop of whole blood - Fast : result within 1 min different examples of glucometers example of use in the dog

10 URINE DIPSTICKS Identification and semiquantitative measurement of several analytes - glucose - ketones - bilirubin - urobilinogen - (specific gravity) - ph -protein -nitrites - (leukocytes) - blood/hb 3-28 SIMPLE TESTS FOR COAGULATION DEFFECTS Whole blood coagulation can be evaluated if «chronometric techniques» are used -Includes: Water bath/heating device + reagents + chronometer (stopwatch) SPECIAL TUBES AND HEATING DEVICE FOR ACTIVATED CLOTTING TIME (ACT) TEST (a whole blood coagulation test) 3-29 SIMPLE SEROLOGICAL TESTS (SNAP TESTS): AN EXAMPLE FeLV / FIV kits Available kits detect Antibodies against FIV Antigen (p27) for FeLV Use serum/plasma. Do not use whole blood Read results ON TIME Snap

11 BASIC EQUIPMENT All veterinary clinics should have at least equipment suitable for specimen preparation specimen storage specimen shipment to specialised laboratories performance of urgent analyses safe working conditions safe disposal of biological material, glass, needles etc 3-31 BASIC EQUIPMENT: SAFE WORKING CONDITIONS - Use a separate room or part of a room - Control of ventilation and temperature - Adequate storage and handling of reagents and specimens 3-32 BASIC EQUIPMENT All veterinary clinics should have at least equipment suitable for specimen preparation specimen storage specimen shipment to specialised laboratories performance of urgent analyses safe working conditions safe disposal of biological material, glass, needles etc

12 BASIC EQUIPMENT: WASTE DISPOSAL - Special separate bins for biological, contaminated, or infectious material, chemicals, etc. - Special bin for needles, blades, glass, etc. - Knowledge of legal issues regarding safe disposal of biological material RECOMMENDED (ADVISED) EQUIPMENT 3-35 RECOMMENDED EQUIPMENT automated equipment for analyses which may be used in practice every day + - ± complete panel of analytical variables Improvement in the quality of analysis compared with basic equipment costs are moderate to high, but affordable if enough samples are processed

13 RECOMMENDED EQUIPMENT Equipment - Automated blood cell counter - Coagulometer Constituents analyzed - Detailed and complete haematology panel - Coagulation parameters - Clinical biochemistry analyser; Ionometer - Multiple biochemical tests 3-37 AUTOMATED HAEMATOLOGY ANALYZERS - Three types: Impedance cell counters Quantitative buffy coat analysers Laser cell counters (more for advanced laboratories) 3-38 IMPEDANCE CELL COUNTERS - Provide: RBC + Hct + Hb + WBC + platelets ± differential cell count in some cases. To validate results, examination of a stained blood smear is advised

14 QUANTITATIVE BUFFY COAT ANALYSIS - Provide: PCV + Hb + WBC and platelets estimation of differential WBC To validate results, examination of a stained blood smear is advised 3-40 COAGULOMETER Provide: -PT (Prothrombin time) -aptt (activated partial thromboplastin time) 3-41 CHEMISTRY ANALYSERS: TYPES - Depending on the reagent used: dry chemistry (ideal for emergency, more expensive per test) liquid / wet chemistry (need technical expertise, cheaper per test) - Depending of their suitability to change techniques: open (good technical skills needed more difficult for routine use in practice) closed (minimal technical skills needed more suited to routine use in practice)

15 CHEMISTRY ANALYZERS: differences between open and closed systems 2 different possibilities «open» systems «closed» Photometers Photometers + Any liquid reagent Stabilized, specially Water-bath designed reagents required ± technical skills 3-43 CLOSED CHEMISTRY ANALYZERS Usually: precalibrated or autocalibrated only basic technical skills needed high throughput of samples possible use dry reagents (use only reagents prepared by the manufacturer) 3-44 OPEN SYSTEM: PHOTOMETER + LIQUID REAGENTS - usually: requires calibration good technical skills required lower throughput of samples can use reagents from any commercial supplier

16 IONOMETER - serum/plasma/blood can be used - autocalibrated/some can be portable - provides ionogram: sodium, potassium, chloride ADVANCED EQUIPMENT FOR REFERRAL LABORATORIES 3-47 EQUIPMENT FOR REFERRAL LABS Highly specialized equipment, designed to have a high throughput and analytical quality. Only affordable by laboratories with a large throughput of samples. Unlimited Price analytical high Time possibilities Skills

17 EQUIPMENT FOR REFERRAL LABS Advanced cell counters 3-49 EQUIPMENT FOR REFERRAL LABS A clinical biochemistry multiple autoanalyzer 3-50 EQUIPMENT FOR REFERRAL LABS Blood gas analyzer

18 EQUIPMENT FOR REFERRAL LABS Cytocentrifuge