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Pigment Information and Courses from MediaLab, Inc.

These are the MediaLab courses that cover Pigment and links to relevant pages within the course.

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Laboratories Individuals

Cerebrospinal Fluid
Other Causes of Xanthochromia

Examples of sources of pigment other than oxyhemoglobin and bilirubin that can cause xanthochromia include: methemoglobinincreased CSF protein (> 150 mg/dL)contamination by skin antiseptic (iodine or merthiolate)

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Chemical Screening of Urine by Reagent Strip
Bilirubin Characterization

Bilirubin, a product of hemoglobin breakdown, is characterized by its yellow pigment. The presence of bilirubin in urine is always abnormal. It is important to note that unconjugated bilirubin cannot be excreted by the kidneys because it is bound to albumin and is not soluble in water. In the liver, bilirubin combines with glucuronic acid through the action of a glucuronyl transferase to form water soluble bilirubin diglucuronide. Under normal circumstances, conjugated bilirubin passes from the bile duct and then to the intestinal tract. Intestinal bacteria reduce conjugated bilirubin to urobilinogen. Approximately half of the urobilinogen is excreted in the feces; most of the other half is recirculated through the liver. A small amount of urobilinogen bypasses the liver and is excreted in the urine.

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Test for Bilirubin

The test for bilirubin on the urine reagent strip is based on the formation of an azobilirubin compound resulting from a reaction of bilirubin in an acid medium with diazotized 2, 4 dichloroaniline. The color of this compound ranges through various shades of tan. Some sources describe the colors produced as shades of tan-to-pink-to-violet. Since other pigments in the urine may influence the test results, this test strip is more difficult to interpret than the others. Colors which are unlike either the positive or negative color blocks on the color chart may be due to the presence of bilirubin -derived bile pigments. Any urine which demonstrates an atypical color on the bilirubin test strip should be tested further. Even a slight change in color should be considered significant since bilirubin is never present in normal urine.

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Confirmatory Testing

Confirmatory testing using an alternative method, such as Ictotest reagent tablets, can be performed when positive results are seen on the dipstick strip, when a red color forms on the strip, or when atypical color changes occur that are the result of bilirubin-derived bile pigments in the urine masking the bilirubin reaction.

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Current Topics in Clinical Microbiology
Enterococcus ID

Illustrated in this composite photograph is the gram stain of Enterococcus species (upper), illustrating gram positive cocci arranged in pairs and short chains.The viridans streptococci generally form gram positive cocci in long chains; the gram-positive cells of S. pneumoniae, on the other hand, may appear similar in gram stain.In the lower right is shown a 6.5% NaCl tube. The turbidity indicates growth, a unique characteristic for Enterococcus species.In the lower left frame is a picture of a bile esculin hydrolysis slant, showing a positive reaction (black pigment) characteristic of Enterococcus species. The streptococci are uniformly negative.Some strains of Leuconostoc species and Pediococcus species can grow in 6.5% NaCl and are bile-esculin positive; however, they should be ruled out because of the negative PYR reaction.

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Clostridium Quad Plate

Key reactions for the identification of Clostridium septicum are shown in the two quadrant plates shown in the photograph.Included in the upper photograph are reactions for milk (casein) proteolysis (12 o'clock quadrant), glucose fermentation, DNAse hydrolysis, and starch hydrolysis respectively reading clockwise.The media in the quadrant plate shown in the lower photograph include gelatin hydrolysis (2 o'clock quadrant) and fermentation of each of mannitol, lactose, and rhamnose respectively, reading clockwise.Milk (casein) hydrolysis Glucose fermentation Key reactions for the identification in the upper plate include no proteolysis of milk, fermentation of glucose (yellow red color along the inoculation streak), positive DNAse (reddish clearing around the streak) and negative reaction for starch. Key reactions in the lower plate include hydrolysis of gelatin, fermentation of lactose (yellow pigment), and negative reactions for mannitol and rhamnose (no pigment).Most strains of C. perfringens hydrolyze starch and produce proteolysins of milk, the key reactions that distinguish C. septicum (negative). Reactions to the other tests do not distinguish between the two.

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Staph aureus Mannitol

The ability to grow in 6.5% NaCl and to produce acid from mannitol was once considered sufficient to identify S. aureus.Although other Staphylococcus species have these characteristics, mannitol salt agar, as shown here, can still be used for epidemiologic studies in suspected cases of S. aureus outbreaks.As shown in the photograph, the presence of colonies indicates the ability to grow on 6.5% NaCl; and, the yellow pigment of the colonies indicates acid production from mannitol.

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S. milleri Biochemicals

The combination of decarboxylation of arginine (red color in the 2nd tube from left compared to the yellow color of the control to its left), the hydrolysis of esculin (black pigment in the esculin agar tube) and the reduction of nitrates to nitrites (red color in last tube on the right) are biochemical characteristics confirmatory for S. milleri (anginosus).

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Electrophoresis
Specimens

Serum and plasma are the most common clinical specimens used for electrophoresis applications. Urine and cerebrospinal fluids (CSF) are also suitable. Other body fluids such as pleural fluid and pericardial fluid are analyzed less frequently. Some specimens require pretreatment before electrophoresis. Low concentrations of proteins normally in urine and CSF are concentrated in order to have enough proteins for detectable separations. Some body fluids require removal of pigments, salts, and other compounds that interfere with electrophoresis or the detection of separated solutes. In molecular diagnostic testing of DNA and RNA, the nucleic acids must first be isolated from the specimen and then purified before separation with electrophoresis.

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Mycology: Hyaline and Dematiaceous Fungi
Illustrated in this photograph is a "green lawn" colony of Gliocladium species. The other hyaline mold that produces this type of colony is:View Page

Mycology: Yeasts and Dimorphic Pathogens
Arrange in sequence the steps that should be taken to make a definitive identification of Cryptococcus neoformans.View Page

Red Cell Disorders: Peripheral Blood Clues to Nonneoplastic Conditions
The peripheral blood smear represented by this field was submitted for hematologic review. The RBC inclusions most likely are:View Page

Variations in White Cell Morphology - Granulocytes
Albinism

Albinism, one of the striking physical characteristics of Chediak-Higashi syndrome, is also thought to be related to the general dysfunction of cells. Albinism has been specifically related to the aggregation of melanosomes, the pigment producing cells in the body.

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White Cell and Platelet Disorders: Peripheral Blood Clues to Nonneoplastic Conditions
The cell bulging with inclusions in the image on the right is most consistent with Chediak-Higashi anomaly.View Page


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