S p e c i m e n C o l l e c t i o n, H a n d l i n g & T r a n s p o r t a t i o n
Quality laboratory results begin with proper collection and handling of the specimen submitted for analysis. Correct patient preparation, specimen collection, specimen packaging and transportation are of vital importance.
Specific specimen requirements for each test are provided in the Test Directory. Submit the quantity specified for each test requested. Tests will be canceled as "QNS" (Quantity Not Sufficient) when the sample volume is inadequate and will be indicated as such on the report. As a general rule, the volume of blood drawn should equal 2-1/2 times the amount of serum plasma required. For example, to obtain 4mL serum or plasma, draw at least 10mL blood. When inappropriate or insufficient specimens are submitted, the laboratory will store them and contact the requesting facility to request that the specimen be recollected. Documentation of this action will appear on the report.
- Complete a Test
Request Form (PDF)
for each patient using a black ballpoint pen. Please print. Record all required patient information.
- Patient name, sex, birth date, social security number and medical record or lab reference number.
- Collection date and time.
- Type of specimen submitted
- Condition of specimen (frozen, refrigerated, room temperature).
- Patient fasting conditions.
- Total volume if 24-hour urine.
- Source of specimen if for microbiology or virology test(s).
- Presumptive clinical diagnosis and any pertinent medical history (onset of illness, recent vaccinations, etc.).
- Complete billing information (patient address and all insurance information) if patient or insurance is to be billed.
- ICD-9 Code for each test.
- Mark box(es) [x] indicating the test(s) requested. For tests not pre-printed on the test request form, clearly print the name and order code for each additional test (see Test Directory) on the bottom of the form.
- Keep bottom copy of form for your records.
- Write patient name legibly on specimen containers with a ballpoint pen. Be sure that name or identifier written on specimen exactly matches name on test request form. Provide patient's social security number on tube for Blood Bank tests. Unlabeled specimens will be rejected.
- Fold completed form in half with patient information facing-out.
- Place labeled specimen(s) in the sealed portion of specimen bag. Place the folded test request form into the outer portion of specimen bag.
- On outside of specimen bag mark the box corresponding to correct temperature at which specimen is to be maintained during transport.
Note: If submitting more than one specimen per patient, and specimens need to be stored and transported at different temperatures, use separate bags and separate test request forms for each temperature type.
Frozen Specimens: Serum or plasma specimens need to be frozen only if specifically stated in the specimen requirement. In these cases, it is essential to freeze the specimen as soon as it is
separated from the cells. Always freeze specimens in plastic tubes unless specifically instructed otherwise. Glass tubes are not acceptable. Lay the tube in the freezer at a
45º angle to avoid tube breakage caused by expansion during freezing. Apply labels to the tube with ends overlapping to prevent detachment of label from tube during freezing.
Submit a separate specimen for each test that must be frozen. Thawed samples are not acceptable for analysis. Indicate if the specimen is plasma or serum on both the
specimen and the test request form.
Specimen Storage: Instructions for storage and shipment of specimens for individual tests are listed under specimen requirements in the
Test Directory. Maintain the specimen at the
temperature indicated for each test until specimens are delivered to the laboratory . Most specimens require refrigeration unless otherwise indicated.
Blood
Specimen Collection
Serum Separator Tubes: For most tests requiring serum,
the Cleveland Clinic Reference Laboratory recommends the use of serum separator collection tubes. However,
refer to the individual
test for specific requirements (Test Directory) since certain tests preclude their use.
When using a serum separator tube, follow these instructions:
- Perform venipuncture as with any other blood collection vehicle.
- Invert the tube gently no more than five times (further inversion may cause alterations in sample integrity).
- Do not remove the stopper at any time. Allow the blood to clot for at least 30 minutes but not longer than 1 hour. Do not centerfuge immediately after drawing blood.
- Centrifuge at 2200-2500 RPM for at least 15 minutes.
- Transfer the clear serum to a labeled plastic vial for transport to the laboratory.
Red Top Tubes: Certain tests require the use of red top tubes rather than serum separators. If serum is to be sent for the test it must be removed from the cells to prevent hemolysis.
- Perform venipuncture as with any other blood collection vehicle.
- Do not remove the stopper at any time. Allow the blood to clot for at least 30 minutes but not longer than 1 hour. Do not centrifuge immediately after drawing blood.
- Centrifuge at 2200-2500 RPM for at least 15 minutes.
- Transfer the clear serum to a labeled plastic vial, using a pipette. Do not transfer red blood cells to the vial.
Certain tests may require submission of the entire, uncentrifuged red top tube. Please refer to the
Test Directory
for specific test information.
Plasma: Plasma contains fibrinogen and other clotting factors when separated from the red cells. Evacuated tubes used to collect plasma specimens contain anticoagulant and
frequently a preservative. An additive in the tube is specified and correlates to the color of the tube stopper. Consult the individual test specimen requirement to determine
the correct additive/tube to use. Indicate that the specimen is plasma on the specimen container and test request form.
Follow the instructions below for specimen preparation.
- Perform venipuncture as with any other blood collection vehicle.
- Invert the tube gently no more than five times to mix blood and additive.
- Centrifuge within 1 hour of collection at 2200-2500 RPM for at least 15 minutes.
- Pipet the plasma into a clean labeled vial. Do not transfer red blood cells to the vial.
Whole Blood: Collect whole blood according to the instructions provided for the individual test. Thoroughly mix the blood with the additives by gently inverting the tube. Maintain the specimen at room temperature before shipping to the laboratory unless instructed otherwise. Never freeze whole blood unless specifically instructed by the specimen requirements. When frozen whole blood is required, transfer the specimen to a labeled plastic vial. Never freeze glass tubes.
Coagulation Testing Requirements
Coagulation Collection To Obtain Platelet-Poor Plasma: For most coagulation tests and factor assays, use the following instructions. For individual requirements, refer to the specific tests in the
Test Directory. In order to produce valid results for coagulation tests and factor assays, specimen integrity is crucial and must be maintained. All specimens sent for testing must be collected and shipped in the
following manner:
- Obtain venous blood by drawing a clearing tube prior to obtaining the specimen. Draw the specimen in a light blue top sodium citrate tube. Avoid stasis and contamination of the specimen by tissue thromboplastin.
- Mix blood with anticoagulant (3.2% or 3.8% buffered sodium citrate) by gentle inversion. Use 0.5mL sodium citrate for every 4.5mL blood. An exact ration of 9 parts blood to 1 part coagulant should be maintained.
- Centrifuge the specimen at 2500 x g for 15 minutes. Hemolyzed specimens may be rejected.
- Immediately remove 2mL (min: 1mL) platelet-poor plasma from the red cells using a plastic pipet. Place the plasma into a properly labeled plastic vial. Glass vials will not be accepted. Freeze immediately. Each assay requested must be submitted in a separate plastic vial.
- Transport frozen on dry ice.
- Some assays may be performed on a priority basis if a medical emergency exists. Contact Client Services to make arrangements.
- All requests for coagulation panels (Hypercoagulation, Von Willebrand's etc.) must include a brief patient history and other pertinent clinical information (i.e., medications, blood products, etc.) Complete the Coagulation Patient History Form and submit with specimen. The interpretation of complex coagulation tests is facilitated by receiving pertinent patient history.
Criteria for Rejection Of Citrated Samples: Citrated samples will be rejected and the tests canceled under the following conditions:
- The specimen is not labeled properly.
- A laboratory accident occurs and the sample is spilled or the tube is broken.
- Abnormal results are found in a hemolyzed specimen.
- The specimen is more than moderately hemolzed.
- A clot is found in the sample.
- The tube is inadequately filled (less than 1/2 full).
- The hematocrit (Hct) of the sample is 50% or greater. In this case, the sample must be redrawn,
using an adjusted whole blood to sodium citrate ratio. Use the
following equation to calculate the amount (in mL) of whole blood to
be drawn in a syringe using 0.5 ml citrate:
60 / (100-Hct) x 4.5 = mL
of whole blood to be drawn. Specimens with Hct > 50% have a correspondingly decreased plasma volume, which results in spuriously elevated PT and APTT results.
HyperCoagulation Testing Algorithm (TIFF, PDF)
Urine Specimen Collection
Random: The normal composition of urine varies considerably during a 24-hour period. Most reference values are based on analysis of the first urine voided in the morning. This
specimen is preferred because it has a more uniform volume and concentration, and its lower pH helps preserve the formed elements.
Submit a first morning specimen whenever possible. Urine for pregnancy testing should be a first morning voiding, or a random specimen with a specific gravity of at least
1.010. Note the time of collection of the specimen on the test request form and on the label of the container. Submit urine for pregnancy testing in a plastic vial with no
preservative. Refer to the Test Directory for specific analyte requirements.
To reduce contamination, the specimen submitted for urinalysis should be a clean catch "midstream sample." Urinalysis specimens must be submitted in a yellow capped urine
transport tube containing preservative. The tubes are available through
Client Services.
24-Hour Urine Collection:
Proper collection and preservation of 24-hour urine specimens is essential for accurate test results. Patients must be carefully instructed in the correct procedure. Refer to the
Test Directory
for specific analyte information. If a frozen specimen is required, freeze the urine aliquot immediately after collection. Pack in dry ice for shipment to the
laboratory. (Refer to section on Frozen Specimens).
Give a collection container and detailed instructions to the patient. If a urine preservative is required, it is important that the designated preservative be in the urine collection
container at the start of the collection. Caution the patient that the preservative may be toxic and caustic and that it should not be spilled or discarded. Record any medication
that the patient is receiving on the test request form. Mix the urine well, measure the volume of the 24-hour collection and record volume on both the test request form and the
transport vial. Transfer the required volume into a plain urine transport vial. Add any additional required preservative and mix well.
Do not send the entire urine collection.
Note: For those analyses requiring the addition of 6N HCI or other preservatives, add the preservative at the start of collection. Have the patient collect each specimen in a
smaller container and carefully pour the urine into the 24-hour container to avoid any possible acid burns to the patient.
Be sure to mix urine thoroughly before removing the aliquot.
- Avoid alcoholic beverages and vitamins for at least 24 hours before starting to collect urine, and during the collection period. Do not discontinue medications unless instructed to do so by your physician. Inform the laboratory which medications you are taking.
- Do not exceed your normal intake of liquids or change your dietary habits during the day before and the day of your collection unless your physician gives you specific instructions to do so.
- Empty bladder (void) into the toilet on the morning of the collection day. Do not include the first urine specimen of the day.
- Collect all subsequent urine voided for the next 24 hours and add to the container provided by the laboratory. The last sample collected should be the first specimen of the following morning at the same time as the previous morning's first voiding.
- Keep the urine in a cool place. Refrigerate if possible.
- Deliver to the referring healthcare provider promptly.
- Preservative may have been added to this container. This may be caustic. Please be careful not to spill any of the preservative.
Stool Specimen Collection
(24, 48
or 72-Hour)
Special gallon-size containers are available upon request for the collection
of stool specimens. Refer to specimen requirements for each analyte.
Flow Cytometry
A full range of services are available for the assessment of immunodeficiency states and the classification of leukemia and lymphoma.
For specific test information, please contact Client Services.
Available Tests and Applications:
-
Immunodeficiency States
-
Immunodeficiency (CDC) Panel
-
Immunodeficiency Basic Panel
-
CD4 Absolute Count
-
Extended HIV FCM Monitoring
-
-
Autoimmune Disease
-
Immune Activation Markers
-
-
Transplantation
-
Stem Cell Absolute Count
-
OKT3/ATG FCM Monitoring
-
-
Oncology
-
Lymphoma Markers
-
CCG Acute Leukemia Markers
-
Low Grade Lymphoma/Leukemia Markers
-
T-Cell Lymphoma Markers
-
nTdT by FCM
-
clgM by FCM
-
DNA Ploidy and Cell Cycle Analysis (Hedley, Vindelov, and Multiparametric Methods)
-
Acute Leukemia Markers
-
Collect whole blood specimens in EDTA, Heparin or ACD vacutainer tubes. Do not dilute bone marrow aspirate specimens with peripheral blood.
Maintain at room temperature. Do not refrigerate.
Deliver specimens to the laboratory within 24 hours. Obtain and transport Monday through Friday only.
Provide the following information on the Test Request Form (PDF)
Patient name, sex and age
Patient Social Security or Identification number
Collection time
Collection date
Sample origin or location
Ordering physician
Send 1 or 2 unstained, air dried blood or bone marrow smears.
Lymph Nodes: Lymph nodes for same-day delivery should be kept at room temperature and submitted wrapped in sterile gauze moistened with sterile saline. Lymph nodes for next-day delivery or longer should be submitted intact suspended in 25mL or RPMI 1640 or SMEM supplemented with 100 mcg/mL gentamicin. These specimens should be refrigerated. Media is available upon request from the Cleveland Clinic Reference Laboratory. A 1 cm3 piece of tissue is optimal. Less tissue may result in a limited analysis.
If a full consultation is requested, submit one of the following:
A large piece of tissue (2 cm3) to allow a portion to be processed for routine histology.
Histological sections and paraffin tissue block (B5, formalin fixative preferred). Indicate that slides/blocks will accompany the flow specimen on the requisition.
Tumors: Tumors for DNA ploidy analysis should be submitted intact suspended in 25mL of RPMI 1640 or SMEM supplemented with 100 mcg/mL gentamicin. These specimens should be refrigerated. Media is available upon request from the Reference Laboratory.
Paraffin blocks of formalin-fixed tissue may be submitted for Hedley DNA ploidy analysis. Hollande's or B5-fixed tissue cannot be analyzed and will be rejected.
Specimen Rejection Criteria: Client Services will notify the ordering facility or healthcare provider of any test cancellations. Listed below are the common causes for test cancellation:
-
Missing patient identification (name/ID number)
-
Missing collection time or date
-
No sample origin
-
Illegible test request form
-
Frozen whole blood specimens
-
Clotted specimens
-
Whole blood or bone marrow specimens not stored and transported at room temperature
-
Insufficient volume (QNS)
-
Age of specimen
Please Note: If specimens older than 24 hours must, by necessity, be processed by the laboratory, the viability must be assessed prior to testing. Any sample indicating <50% viability will be rejected. Specimens indicating >50% viability will be evaluated and the final report will include a viability report. Reports for specimens with a viability range of 50%-85% will contain appropriate disclaimers. An additional fee will be charged for such viability studies.
Questions regarding appropriate handling of specimens are welcome (particularly prior to anticipated biopsy) and may be directed to the Flow Cytometry Laboratory 216.444.0042.
- Serum Separator Tubes
- Red Top Tubes
- Plasma
- Whole Blood
- Coagulation Testing Requirements
- Hypercoagulation Algorithm
Stool
Specimen
Flow Cytometry:
Coagulation Testing Info. & Algorithms
