Blood Collection and Handling of Blood Samples

Collecting your Sample Determine which ________________ are needed. Determine the __________________ you will need and the _______________ you will use. Preferred blood source is almost ALWAYS _______________ blood, not ______________ blood. ___________ vein is usually most appropriate vessel for collection. Use the _________________needle that the patient can comfortably accommodate. Choose the size syringe that best matches the ___________________________________ you will need.

Needle and Syringe

The Vacutainer Is composed of a ______________, _________________________and _________________tubes. Use the correct size tube to minimize _______________of the sample and to prevent ____________________ of the vein. Fill tube to correct volume based on strength of ______________________________ to ensure appropriate ratio of blood to anticoagulant. ADVANTAGE: multiple samples can be collected directly into tubes without __________________________________ from patient.

Vacutainer collection sets

Sample Volume The amount of blood collected from an animal depends on the amount of _______________or _______________needed as well as the ________________________ of the animal. Enough blood should be taken to run the required tests _______ times. This should be enough to compensate for technician error, instrument error or the need for diluted sampl es.

Serum or Plasma? Serum or plasma are the _____________ portion of whole blood. Fluid portion of blood is ____% water, ____% dissolved constituents like proteins, vitamins, carbs, hormones, etc… Plasma ______contain clotting factors. The clotting factors are known as __________. ________is plasma that has had the clotting factors removed.

Whole Blood Is placed into a container with an _____________________added to prevent clotting. As soon as you obtain your sample, mix the blood with the anticoagulant by using a ____________________________ motion. Vigorously shaking your sample can cause ______________, otherwise known as cell destruction.

_____________ Defined: Are chemicals that prevent or delay the clotting process. Choice of anticoagulant depends on _____________needed. Sample must be _______________before use.

Anticoagulants cont’d Samples not tested within _________of collection should be refrigerated. (Bring sample back to room temperature and re-mix before analysis. ___________ blood should NEVER be frozen as the freezing/thawing process can _______ the blood cells.

What’s in the tubes?

Red Topped Tube Red Topped Tube: Contains no ____________________. Routinely used for ________________________. Used for ________samples.

Red-Topped Tubes

Serum Separator/Tiger Topped Tube Tiger Topped (Striped) Tube/Serum Separator: Contains no anti-coagulant. Has a yellowish “plug” of __________________________________that separates serum from plasma when spun. Used for ___________ samples. ***(Not used for therapeutic drug level monitoring.)

Lavender/Purple Topped Tubes Lavender Topped Tube: Contains the anticoagulant __________ or Ethlenediamine tetraacetic acid. Used for ____________________samples or __________samples. Used for complete blood counts because it does not ____________________________. HOWEVER, an excess of anticoagulant in a sample may cause cells to __________and invalidate cell counts done on automated analyzers.

Lavender/Purple Topped Tubes

Grey Topped Tube: Contains the anticoagulant _________________________. Best for ___________________ preservation. Interferes with many other tests performed on serum. Interferes with many other tests performed on serum.

Grey Topped Tubes

Blue Topped Tube Contains the anticoagulant _______________________. Commonly used in ___________________. Na Citrate interferes with Na assays and many common serum tests.

Blue Topped Tubes

Green Topped Tube Contains the anticoagulant ________. Can be used for most tests that require ____________samples. Should never be used for differential blood film analysis because the anticoagulant interferes with the staining of the _______’s.

Green Topped Tubes

Hematology Defined: ____________________________

Why is hematology important? Evaluation of _________________ Screening for _______ animals as a _______________. ___________________ screening _______-___________ monitoring

Hematopoiesis Refers to the production of __________________________ and __________________. All blood cells arise from the same _______________________________________.

Blood Composition Blood is composed of __________and __________ Fluid portion is ~____% water

Packed Cell Volume The PCV is measuring the percent of CELLS in a patient’s blood. If the animal is dehydrated, the fluid portion of the blood will ____________. Example: a PCV of 50% will give a sample that is 50% cells and 50% fluid. This means that a 10mL sample will yield 5mL of fluid. A PCV of 70% will yield 70% cells and only 30% fluid so a 10mL sample will only give 3mL of fluid.

PCV (Packed Cell Volume) In a CBC, we determine the number of RBC’s in several different ways. The quickest and easiest is called the __________________, also referred to as the packed cell volume (PCV) The PCV will tell you if the animal is ________or ________.

Normal PCV Values Canine: _______% Feline: _______% Equine: _______% Bovine: _______%

Whole blood is collected in an ______________ (usually EDTA) and placed in a capillary tube. Microhematocrit tubes should be filled to the designated line, with one end plugged with clay sealant.

Blood sample should be spun in a microhematocrit centrifuge for ______ minutes Lay the tube in the centrifuge with the plugged end facing the ___________of the centrifuge. Make sure that a _______________is placed opposite or have another sample across from yours. Cells are ________than the plasma and are compacted at the end of the tube that has the clay plug.

Plasma Evaluation Plasma color and transparency may be helpful in determining a diagnosis and should be recorded in your findings. Normal plasma is _____or a _____________color Cloudy Plasma = ________ Reddish tinge = _________ Yellow = ________(indicates possible liver disease)

Reading your PCV

Concentration of total protein / total solids Plasma protein concentrations estimated with a _____________and is an important component of the CBC in all species. Plasma used to determine the TP/TS is collected by breaking the hematocrit tube just above the _________/_________interface. ↵

A perfect PCV specimen here at VTI

The plasma is allowed to flow onto the ______________________________________. (Blow gently through the open end of the hematocrit tube with the broken end of the tube over the prism of your refractometer.) Hold the refractometer up to the light and record your findings. Make sure to ____________your refractometer after each use!

Blood Films The blood film is used to perform the ____________, estimate platelet numbers; and evaluate the _________________________________features of WBCs, RBCs and platelets. _________________ smears are prepared by placing a small drop of blood on a clean glass microscope slide

Blood films

Staining a slide Always stain using the ________to _______stain. Remember which side of your slide is up (clothes pins are marked “top”) Rinse off from _____________side of slide May _______________ to speed up process. (We will NOT be doing this!)

Performing the Differential Cell Count This is where the different _____________________________________ are tallied separately. This can be done by a blood counting machine, or by hand. To ________count the different cells, first you must make a ____________. Stain the slide once it is dry. Using a cell counter you will tally a total of _____cells (this will make it easy to turn the numbers into a %)