Assessment Information and Courses from MediaLab, Inc.

Specimen rejection criteria established by your laboratory should be followed at all times, as improperly collected or processed coagulation specimens could adversely affect patient results. Generally speaking, hemolyzed specimens should not be used in coagulation testing because ADP liberated from lysed red blood cells can interfere with a number of coagulation tests, especially those involving platelet assessment. Grossly lipemic specimens may cause erroneous results or a clot may not be detected if a photo-optical coagulation system is used. An alternative method that is not affected by lipemia, such as an electromechanical method, may be required One way to avoid a grossly lipemic specimen is to ask the patient to fast prior to specimen collection.

The activated partial thromboplastin time (APTT) is a screening test that helps to assess the functionality of both the intrinsic and common pathways. The effectiveness and presence of all the coagulation factors are assayed by this diagnostic test with the exception of factors VII and XIII. The results of the activated partial thromboplastin time are used in conjunction with other diagnostic tests, as well as the clinical picture of the patient, to determine hemostatic abnormalities which may be present. In addition to being an integral part of the coagulation disorder assessment process, the APTT is used to determine therapeutic effectiveness of heparin administration. Activated partial thromboplastin time results are presented to the clinician in seconds- the actual time elapsed until a clot was detected using the laboratory's instrument/reagent system.

A covered entity may use or disclose PHI, without getting an individual's authorization, in order to:Perform requested tests and treatments.Bill for the services performed.Perform essential operations, including quality assessment, accreditation, and compliance.Meet legal reporting requirements, including those mandated by public health departments, workers' compensation, law enforcement agencies, and the US Department of Health and Human Services. Other uses and disclosures require written authorization.

Recent events, including the terrorist attacks on September 11, 2001 and the subsequent bioterrorist releases of anthrax, have been a harsh awakening that the nation’s workplaces could be terrorist targets.Traditionally laboratory safety guidelines have emphasized use of optimal work practices, appropriate containment equipment, well-designed facilities, and administrative controls to minimize risks of unintentional infection or injury for laboratory workers. Today, in addition to the above, laboratories must make a risk and threat assessment, secure data and electronic technology systems, plus develop policies regarding specimen accountability, facility security, and emergency response.The next few pages will cover a number of things that you can do to assist in making your laboratory more risk free to a terrorist attack and some things you can do in case that security is breached. You too have a role in the security of your workplace!

Particle smears are also made from the unanticoagulated sample. The bone marrow particles are removed from the watchglass and placed on a coverslip. One of the following items: Pasteur pipet, capillary tube or broken end of a wooden applicator stick, may be used to transfer the particles. A second coverslip is placed over the first and the particles are crushed between the coverslips as they are pulled apart. Some practice is needed to perfect this technique. As mentioned previously, this type of preparation provides a more accurate assessment of marrow architecture and cellularity than the direct smear. Morphological detail is preserved on well made slides. The remaining sample may be added to a tube containing EDTA anticoagulant and additional smears may be made if needed.

In order to discuss TDM and PGx we need to also introduce the concept of pharmacokinetics. Pharmacokinetics is the study of drug disposition in the body: how and when drugs enter the circulation, how long they remain in the blood, and how they are eliminated. TDM is the clinical assessment of a drug's pharmacokinetic properties. Physicians and pharmacists need to establish that a drug is present at an effective concentration but not at a toxic concentration. The next few pages will describe some of the factors that determine a drug's disposition in the body. These factors ultimately decide the need for therapeutic drug monitoring.

Linear calibration curves are more desirable because they result in the best accuracy and precision. Some testing methods, however, do have nonlinear calibration curves. When that is the case, more calibration standards are needed to achieve desirable precision. Regardless, linearity is not to be used as a tool for calibration verification, accuracy assessment, or establishing a reportable range.

These cells are in an area which is too thick, and should not be used for red cell morphology assessment. Some of the cells appear to be stacked like coins because of the large number of cells present in this section of the slide. The morphology seen in the too thin and too thick areas of the smear is referred to as artificial morphology.

The following aspects of semen analysis will be described in further detail during this course: Check the identity of the patient Record information that has been obtained from the patient including: time of collection, collection method, problems during collection, medications the patient is taking Note time to liquefaction Measure the volume by pouring into a graduated test tube or by drawing the specimen into an appropriately sized graduated serological pipet Assess viscosity Note color Measure pH by putting a drop on a strip of pH paper Count the sperm in the specimen Assess motility Count round cells, if present Assess the proportion of round cells that are white cells Fix and prepare specimen for morphology assessment; assess morphology

Describing the morphology of the sperm in a semen specimen is an essential part of the microscopic examination. The presence of abnormal forms along with low counts and/or poor motility contributes to a poor prognosis in infertility cases. There are several different methods for determining morphology. The most common are the WHO III (WHO III manual, 1992)assessment and the Strict Morphology method found in the WHO IV manual (1999). A specimen is considered normal if 30% or more of the sperm are normal morphology according to WHO III criteria. If strict morphology criteria are used then the specimen is considered normal if it has 14% or more normal forms.

Viability is a measure of the percentage of cells that are alive. Since motile cells are inherently viable a viability assessment may not be necessary when motility is high. Most laboratories set a minimum motility after which a viability will also be performed. To assess viability, place a drop of semen on a slide. Add an equal volume of a vital stain such as trypan blue. Cover with a coverslip. Allow color to develop for several minutes, but not more than 5 minutes. Count 100 cells (both motile and non-motile cells) on each of 2 slides. During the count differentiate between the white cells (living) and blue cells (non-living).

Sperm can be counted either manually or by automated methods. Although automated counting has some advantages for assessment of motility parameters, manual counting is still performed by most laboratories. There are several manual counting methods available for semen. These include:Neubauer hemacytometerMakler chamberCellVu (Millennium Sciences, Inc)MicroCell (Conception Technologies) The Makler, CellVu, and MicroCell methods have the advantage of requiring no dilution of the semen. Since semen is viscous, accurate dilution can be problematic. These methods also allow counting of motile and non-motile sperm at the same time and thus avoid the need for separate assessment via wet mount. Each laboratory should determine the best most reproducible method for their own situation, equipment, and expertise.

To verify the absence of sperm after vasectomy, two successive negative specimens are ideally required. If no sperm are seen on the initial undiluted count, the specimen should be centrifuged to concentrate any cellular material, and re-examined.

For assessing count, motility, viability and other cellular components your laboratory will require a microscope with 10x oculars and phase contrast objectives up to 20x. You will also need hand counters.For assessing morphology you will need bright field objectives of 40x and/or 100x (oil immersion).You will also need counting chambers, glass slides and coverslips and a method for staining sperm for morphology assessment.

Respirators are used in situations that pose a high risk for exposure.Respirator usage for TB is now regulated under the general industry standard for respiratory protection.Risk assessment determines HCWs who should wear respiratory protection.HCWS are screened for medical conditions by a physician prior to using respiratory protection.Respirators should be selected from those approved by CDC and NIOSH.Fit testing provides a method to determine which respirator model and size fits the wearer best and to confirm that the wearer can properly fit the respirator. Each time the respirator is worn, the wearer performs a user-seal check to ensure adequate respiratory protection.

Illustrated in the photograph is a normal bone marrow smear stained with Wright/Giemsa stain. Note the evenly distributed cells with normal maturation in both the myeloid and erythroid maturation sequences.An estimation of the percentage composition of cells can be made by experienced observers from scanning of multiple fields. In some instances a detailed differential count of 300 or more cells must be made.In normal bone marrows, the myeloid to erythroid ratio (M:E ratio)ranges from 1.2:1 to 5:1.A ratio of less than 1.2:1 indicates depressed leukopoiesis or erythroid hyperplasia. Ratios of 6:1 or greater usually indicates infection, erythroid hypoplasia, or chronic myelogenous leukemia.An assessment of the overall cellularity is also useful. In general, cellularity of less than 25% indicates hypoplasia; greater than 75% indicates hyperplasia.