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What is pathology?

The study of disease

Disease

Any variation from the normal morphology or physiology of a living organism.

This can result from various causes (infection, genetic defect, environmental stress, etc.


Characterized by an identifiable group of lesions, clinical signs, or symptoms.

Lesion

Anatomically distinguishable changes in the body tissues, both gross and microscopic.

Clinical signs

Objectively observable changes in behavior or function.

Symptoms

Subjective changes in behavior or function.

Diagnostic pathology

Discipline aimed at diagnosing a disease.

Anatomic

Strives to diagnose disease by concentrating those anatomical (morphological) changes in living tissues (lesions) at the gross and microscopic levels.

Clinical

Strives to diagnose disease by the use of tests on various body fluids and body waste products (i.e. blood, plasma, urine, CSF, sputum, saliva, peritoneal fluid, thoracic fluid, feces, etc.)

The Veterinary Practice Lab

-Dedicated area for lab equipment and running tests


-May be a seperate room or counter top


-Should have adequate storage space


-Convenient electrical outlets


-Nearby sink


-Nearby safety equipment

Biohazard label

-Denotes the presences of potentially hazardous biological material---> use caution


- May be contaminated equipment or biological tissues


-NOT radioactive


- May be present on bags or containers, or designate an entire area as biohazard

Lab Safety Equipment

- Chemical drug vent


- Eye wash station


- Sharps container


- Fire extinguisher


- First aid kit

Chemical drug vent

Used to mix and draw up chemotherapy

Eye wash station

- Used to flush eyes if exposed to harmful materials


- Passive water pressure design


- Water/saline solution

Sharps container

- Used for disposal of sharl materials; primarily syringe needles and occasionally glass slides


- Made of stong punture-proof plastic


- Once full, it is disposed off by special bio-waste handling companies

Fire extinguiser

- Uses either pressurised dry chemicals, or carbon dioxide


- Helps to extinguish a fire

First aid kit

- Contains basic first aid materials in case of an emergency

A clinical pathology/parasitology lab generally requires what type of equipment and supplies?

- Microscope (stereoscope/dissecting microscope, light microscope, electron microscope)


- Centrifuge


- Refractometer


- Hemocytometer/Unopette Pipette


- Differential counters


- Chemistry analyzer/Automated Blood Cell Counter


- Fecalyzer cups


- Agglutination cards


- Refrigerator


- Staining solutions and Supplies

Stereoscope/dissecting microscope

- Useful in examining objects thats can be seen with the naked eye but also requires magnificationfor detail


•Often used to view external parasites like fleas, ticks, and lice


• Uses overhead ("reflective") lighting


• Objects viewed in 3-D

Light microscope

- Primary lab equipment for viewing microscopic objects.


- Most are binocular design


- Most commom objectives: 4x, 10x, 40x, 100x



• 4x = scanning


• 10x = low power


• 40x = high dry


• 100x = oil immersion



- Most use transmitted ("underneath" lighting)


- Eyepiece lenses (oculars) are always 10x


- Oculars are always 10x so the image total magnification is 10 times the objective power

Electron microscope

- Not used in veterinary office setting


- Uses electrons to view image


- Images are always black and white (grayscale)


- Images may be artificially colorized

Centrifuge

- Seperates cells and particulate matter from the fluid in which they are suspended


-Supernatant


-Sedement

Supernatant

Centerfuged fluid

Sediment

Particular matter

What are three types of centrifuges?

1) Horizontal head


2) Angled centrifuged head


3) Microhematocrit centrifuge

Refractometer

- Measures a fluid's specific gravity or protein concentration by measuring how the fluid refracts or bends light


• Blood plasma--> determine protein concentration


• Urine--> determine specific gravity


- Calibrate using distilled water

Hemocytometer/Unopette Pipette

- Convenient and inexpensive way to count blood cells


- Special microscope slide with precisely etched grids within a counting chamber


- Holds an exact volume of diluted blood sample


- Involves physically counting the number of cells in several grid squares and taking an average


- Final result expresses the number of red blood cells per cubic millimeter of the original blood sample

Differential counters

- Special “differential counters” are used when evaluating a blood smear


- Counter helps keep track of the different types of white blood cells observed

Chemistry analyzer/Automated Blood Cell Counter

- Several types are available for veterinary offices


- Perform basic clinical chemistry analysis and blood cell counts


- Machines require numerous specialized reagents and supplies to monitor quality control to ensure accurate results

Fecalyzer cups

- Specialized solution and cup used to examine animal fecal material


•Examined under the microscope for evidence of parasitic eggs (ova) via fecal floatation method

Agglutination cards

- Brand name: Rapid Vet-H Blood Typing card


- Used to determine the presence of certain antigens on the surface of an animal’s red blood cells (blood typing)


- Specialized cards contain antibodies used to test blood types: DEA 1.1 positive and negative dogs


- A positive reaction = agglutination

Refrigerator

- Most laboratories need at least a small refrigerator for reagents and samples that require refrigeration


- Some are specialized, allowing control of not only temperature but other parameters such as humidity


- Because of the biohazard nature of the contents of the refrigerator, these should be used EXCLUSIVELY for lab materials, biological samples, reagents, etc.

Stains

- Wright stain (Romanowsky, Diff-Quik)


- Urine Sedi-stain


- Gram stain

Supplies

- Plastic bulb pipettes


- Kimwipes


- Lens paper


- Distilled water


- Isopropyl alcohol


-Triple 2 cleaner

Microscope use

- Use both hands when lifting or moving the microscope



- Scopes are heavy an expensive



- Grasp the microscope by using one hand under the base and the other on the arm/pillar



- DO NOT grasp the microscope by the stage or the oculars



- Once you place the scope on the table, refrain from moving it



- If you have to move it, pick it up. Avoid dragging the scope on the table as the vibration may cause it to bounce and damage the optics.



- Make sure the microscope light switch is turned off and the light dimmer knob is at the lowest setting- Plug the microscope into the electrical outlet nearest you.- Grasp and rotate the nosepiece to swing the 4x objective lens in place- Gently lower the stage by using the coarse adjustment knob


- Get a slide and place it in on the stage and make sure it is securely positioned by the ‘swing lever’- Center your slide over the stage condenser lens by moving the stage control knobs- Turn the power switch on- Adjust the distance between the binocular lenses to fit your eyes- Look into the binocular lens •If you do not see anything (darkness), check to see if the light is on and the light intensity is bright enough



- While looking through the binocular lens, use the coarse adjustment knob to slowly raise the stage until the objects on the slide start to become focused.



- Use the fine adjustment knob to bring the objects on the slide into complete focus



- Get a slide and place it in on the stage and make sure it is securely positioned by the ‘swing lever’



- Center your slide over the stage condenser lens by moving the stage control knobs



- Turn the power switch on



- Adjust the distance between the binocular lenses to fit your eyes



- Look into the binocular lens



•If you do not see anything (darkness), check to see if the light is on and the light intensity is bright enough



- While looking through the binocular lens, use the coarse adjustment knob to slowly raise the stage until the objects on the slide start to become focused.



- Use the fine adjustment knob to bring the objects on the slide into complete focus



- Move the slide using the stage control knobs until you have viewed all of the material on the slide



•Oil immersion- Make sure you identify which objective is labeled “oil”



- Move the slide using the stage control knobs until you have viewed all of the material on the slide



-NEVER use oil on the non-oil objectives- Lower the stage to give adequate room for applying oil- Also rotate the objective nose piece so the oil objective is next



- Place one drop of oil directly on the specimen- Rotate the oil objective lens into place- Raise the stage until the objective contacts the oil drop (directly visualizing the oil-objective contact)



- Look through oculars and focus manually and focus normally- When finished, clean oil off objective lens with lens paper and alcohol


Glass slides

Most glass slides have a frosted area on one end. This frosted area is for labeling the slight usually in pencil. When placing the specimen on the a slide, the frosted side should be up

Laboratory Quality Control

-Quality control (QC) or quality assurance: process for assessing the accuracy and reliability of a test system


•Any clinical laboratory should adhere to a comprehensive program to ensure lab data reliability •Usually accomplished by running specimens with known values or outcomes along with patient unknowns


•QC programs are generally designed for that specific laboratory, but at minimum should address:


▪Requirements for staff proficiency


▪Equipment maintenance and monitoring


▪Other standards of operations


•A written QC policy should include:


▪The control materials to be use with each assay


▪Frequency of use


▪Required documentation as well as criteria for the acceptability of patient results