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Budding Information and Courses from MediaLab, Inc.

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

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Body Fluid Differential Tutorial

The white cell clump in this image contains budding yeast and matted branching hyphae contained within it.If you look away from the clump, the yeast and hyphae are not obvious. This is why it is important to always take a close look at any white cell clumps that are present on a cytospin. It is good practice to check fluid differential cell clumps for the presence of infectious organisms.

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Chemistry / Urinalysis Question Bank - Review Mode (no CE)
The elements indicated by the arrows are more likely to be seen in patients with which condition:View Page
Identify the urine sediment elements shown by the arrow:View Page
Identify the urine sediment elements shown by the arrow:View Page

HIV: Structure and Replication (retired 2/20/2013)
The stage of viral replication where the envelope is being acquired and the HIV is leaving the host cell is known as:View Page
Spread of the Infection (3)

As the envelope is being formed, the HIV leaves the cell. This stage is known as budding. The virus moves through the cell membrane, acquires an envelope, and exits into the extracellular environment. It is now ready to infect another cell.

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Destruction of the Host Cell

The mechanism of host cell destruction remains under investigation, but three possibilities have been proposed:Viral budding tears a hole in the host cell's membrane, causing cytoplasmic leakage.The supply of useful T-lymphocytes is depleted, resulting from the formation of huge, functionless syncytia.Some mechanisms cause the provirus to provoke the synthesis of a large number of new HIV particles, causing the rapid depletion of cellular components and with it the destruction of the cell.

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Mycology: Yeasts and Dimorphic Pathogens (retired 2/12/2013)
One of the characteristics common to the dimorphic molds is the ability to convert the mold forms to the yeast forms by incubating subcultures in enriched media at 35°-37°C. The upper image illustrates a subculture of a mold colony suspected of being a dimorphic fungus inoculated to the surface of blood agar and incubated for 3 days at 37°C. Note that the colonies have a prickly appearance, suggesting an intermediate stage of conversion. The lower image is a lactophenol blue mount of a portion of one of the prickly colonies. This fungus can be identified as:View Page
The colonies growing on the surface of this brain-heart infusion with blood agar plate were "converted" from a mold colony suspected of being Histoplasma capsulatum by incubating a subculture at 37°C for 5 days. The yeast forms that must be identified in mounts made from one of these colonies to confirm the identification are:View Page
Match each of the microscopic structures listed in the drop-down box with the name of its corresponding yeast species on the right.View Page
Match each of the fungal species listed below with the appropriate category, indicating whether or not it has the capability of producing pseudohyphae on cornmeal agar.View Page
The colony shown in the upper image was recovered from peritoneal fluid of a patient receiving continuous peritoneal dialysis. The lower image is a photomicrograph prepared from a small portion of the colony illustrating the microscopic morphology. Each of the following species of yeast can be eliminated except:View Page
This photomicrograph is a representative field of a Wright-Giemsa-stained bone marrow aspirate in which a pair of budding yeast cells is seen centrally (arrows). Based on the appearance of these yeast cells, what other test would you expect to be positive?View Page

Reading and Reporting Gram Stained Direct Smears
Size and Appearance of Nonbacterial Cellular Elements on Gram Stained Smears

Type of Cell Average Size Image Comments Epithelial cells 25 µm Appear pink/red on Gram stained smear. Larger than white blood cells. Have a single nucleus. They are an indication of a suboptimal or unacceptable specimen if present in large numbers in sputum specimens, tracheal or endotracheal aspirates, or in urine specimens. White blood cells 12 µm Appear pink/red on Gram stained smear. Most often, polymorphonuclear white blood cells (PMNs). White blood cells indicate inflammation and possible infection. The direct smear examination should focus within and around these cells. Hyphae/pseudohyphae Varies Appear blue on Gram stained smear. Hyphae are tubular filamentous fungal elements, which may show branching or intertwining. Pseudohyphae are multiple buds of yeast that do not detach, thereby forming chains. Yeast 7 µm Appear blue on Gram stained smear. Round to oval, often budding. About the same size as red blood cells. Generally much larger than bacteria. A few yeast may be present as normal flora in upper respiratory tract or genital tract. They may be significant if they predominate, or if budding yeast forms are seen. Red blood cells 7µm Appear red on Gram stained smear Not usually considered a significant finding.

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Reporting Microscopic Findings

Gram-stained direct smears are examined using the oil immersion objective of the microscope (total magnification =1000x). The quantity and type of bacteria and nonbacterial cellular elements present are recorded. A minimum of ten microscopic fields should be examined before reporting the Gram stain result. Organisms and other cells that are observed on a Gram-stained smear should be reported with as much description as possible.In addition to staining reaction, the shape of the organisms should be reported (e.g., cocci or bacilli). It may also be useful to report the cellular arrangement of microorganisms on the smear, although this is more often reported from a smear that is made from culture rather than a direct smear. Cellular arrangements may be described as: Single cells Pairs Tetrads Chains Clusters Budding (in the case of yeast)

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Reading Gram Stained Direct Smears
Yeast cells

Yeast cells are approximately the same size as red cells but stain Gram positive. They appear oval and budding.

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Size and Appearance of Cellular Elements

Type of Cell Size Shape Stain Epithelial cells 25 microns irregular pink/red White blood cells 12 microns round pink/red Red blood cells 7 microns round pink/red Yeast 7 microns ovoid blue Bacteria 0.5 - 1 micron variable blue or red Epithelial cells are larger than white blood cells and red blood cells, and contain a single nucleus. White blood cells (neutrophils contained in pus) usually show a segmented nucleus. Red blood cells are 1/2 to 2/3 as large as white blood cells, contain no nucleus, and are Gram negative. Hyphae are gram positive tubular filamentous fungal elements which may show branching or intertwining. Yeast cells are round to oval, often budding, Gram positive fungal elements, about the same size as RBCs. They are generally much larger than bacteria.

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Significance of Specific Findings:

Epithelial cells in large numbers within sputum smears means that the specimen is predominantly oral saliva, rather than true sputum from the lung. Epithelial cells in urine smears indicate that the sample has been contaminated by organisms found on the vulva or distal urethra. Bacteria found near or on epithelial cells are usually normal contaminating bacterial flora.White blood cells indicate inflammation and possible infection. The direct smear examination should focus within and around these cells.Red blood cells in a direct smear are not usually significant.Yeast may be present as normal flora in upper respiratory tract or genital tract. They may be significant if they predominate, or if budding yeast forms are seen.Hyphae are more likely to indicate the presence of fungal infection, but this determination requires correlation with clinical findings.Bacteria found in spinal fluid, blood, tissue and specimens from other sterile sites are always significant.Body fluids which are normally sterile must be examined carefully. If only one organism per oil immersion field is identified, then there are about 105 organisms per mL present in the sample! Bacteria observed in specimens from the throat, genital tract and other areas containing normal flora suggest infection only if their composition and type varies significantly from the norm.

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Reading Gram Stained Smears From Cultures (retired 2/12/2013)
Which of the following appear as budding organisms?View Page
The ends of rod-shaped bacteria may appear:View Page
Select all the choices below than correctly describe this Gram stained smear.View Page
Select all the choices below than correctly describe this Gram stained smear.View Page
Which of the following terms can be used to describe cocci?View Page

The Urine Microscopic: Microscopic Analysis of Urine Sediment

Yeast can appear as single cells or in the budding form. As single cells they can be confused with RBCs because they are about the same size. In the budding form, yeast is easily identified as demonstrated on this slide. Yeast can be found in patients with cystitis due to yeast, usually candida, or as a vaginal contaminant from patient's with vaginal candidiasis.

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Which of the following are characteristic of normal RBCs under high power (400X magnification) brightfield microscopy? (Choose all that apply.)View Page

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