Microscopic UA: Part 1, 2, 3 Flashcards
Describe the recommended methods for standardizing specimen preparation, volume, centrifugation, sediment preparation, volume and examination, and reporting of results.
Ask Caroline
State the purpose of Sternheimer-Malbin
delineates(portrays) nucleus and cytoplasm. Used for WBC, epithelial cells and casts (purple).
-It is a general purpose stain. The MOST COMMON stain for MICROSCOPIC sediment.
This stain consist of CRYSTAL VIOLET and SAFRANIN O.
True or False: Sternheimer-Malbin or Sedi-stain will not stain fat cells.
It will only stain specific elements and this is not one of them.
State the purpose of acetic acid
-Used to dissolve amorphous phosphates.
-Used to enhance nuclear detail of WBC
-WILL LYSE RBC’s, but not yeast or bacteria.
-Used to differentiate RBC from WBC, yeast, etc. (dissolves WBC granules) and to dissolve amorphous phosphates (to see bacteria).
State the purpose of toluidine blue
delineates nuclear detail. Used to differentiate WBC from RTE (blue).
Toluidine blue helps us see more clearly the inside of WBC from the inside of Renal tubular epithelial cells.
Urine specimens that contain a large amount of mononuclear cells (except for epithelial cells) may need to be referred to cytology. —-> Difficulty may arise when differentiating WBC’s from mononuclear cells (including RTE’s). RTE’s are larger than WBC, but not as round, and the nucleus is usually prominent.
State the purpose of Sudan III and oil red O
Lipid stain
-Used to stain triglycerides and neutral fat.
-Used to identify fat except for cholesterol (reddish color)
It differentiates lipids like Oval fat bodies. In an unstained urine oval fat body will like like clear small cells all clumped up together. When we apply the stain it will stain red and that will help us confirm lipids.
True or False triglycerides and neutral fats stain orange-red and cholesterol stains yellow.
False. Triglycerides and neutral fats will stain orange-red, but cholesterol will not stain.
We will need to use polarized light to help in identifying cholesterol. In polarized light, cholesterol will have those characteristics of Maltese-cross formation. It looks like for ovals and a diamond in the middle.
State the purpose of Gram in the examination of the urine sediment
In urinalysis it is used mainly to identify BACTERIAL casts (can be confused with granular casts).
Many granular casts’ appearance is very granular that it’s difficult to determine if its bacteria that is causing the appearance or if it is just things in the filtrate that got caught, that it is causing the granular appearance.
So if we are concern that there’s an infection in the tubular itself, we might want to do a Gram stain to double check.
State the purpose of Hansel stain
Used to identify eosinophils from other WBC.
-Used to identify eosinophils in urinary sediment (reddish)
–
WBC’s seen in urine are usually neutrophils associated with microbial infections (not eosinophils). —–> In allergic reactions (drug-induced, etc) producing inflammation of the renal interstitium (inflammation within the nephron), eosinophils will be present.
-It is the most common stain done in urinalysis outside that of steinheimer stain.
State the purpose of Prussian blue stains in the examination of the urine sediment
stains structures containing iron
-Used to identify hemosiderin (blue) granules
Unstained hemosiderin granules are a yellow-brown color (seen in renal tubular epithelial cells and casts after hemoglobinuria). Once they are stained the hemosiderin granules will be stained blue.
–> Side note: they actually look like tiny undefinable (not set shape or size), but differently we will see them with RTE cells.
Describe the basic principle of bright-field and their relationship to sediment examination
-routine for urinalysis
-basic principle is that objects/specimens appear dark against a light background.
–
Sediment constituents with a low refractive index will be overlooked when subjected to light of high intensity. Therefore, sediments must be examined using decreased light controlled by adjusteding the rheostat on the light source, not by lowering the condenser. Staining of the sediment also increased the visualization of these elements when using bright-field.
What is the most common type of microscopy?
bright field
Describe the basic principle of phase-contrast and their relationship to sediment examination
-enhances low refractive objects (such as hyaline casts, mucous, etc).
-the basic principle is that when light passes through cells, small phase shift occur, which are invisible to the human eye. In a phase contrast microscope, these phase shifts are converted into changes in amplitude, which can be observed as differences in image contrast.
–
A lot of hospitals use these!
Describe the basic principle of polarizing and their relationship to sediment examination
-used to identify cholesterol and crystals
Describe the basic principle of interference-contrast microscopy and their relationship to sediment examination.
-used to enhance detail (three dimension)
-basic principle is that it involves two coherent beams of light (from the same small light source) and image contest achieved with gradients in optical path. It produces clear optical sections of which transparent specimens and a 3D shadowed image.
–
not very common in hospital labs
Differentiate between normal and abnormal sediment constituents
Normal :
- 0-5 WBC/hpf
- 0-2 RBC/hpf
- less than 0-2/hpf of transitional epithelia cells
-0-2/hpf of Renal tubular epithelial cells
Abnormal:
-Eosinophils (more tha 1%)
-Bacteria
->2/hpf of RTE
Check with Caroline
Identify urine artifacts :
What is number 1?
Talcum Powder
Identify urine artifacts :
What is number 2?
Starch
Identify urine artifacts :
What is number 3?
Air
-Highly refractile
Identify urine artifacts :
What is number 4?
Oil
-Highly refractile
Identify urine artifacts :
What is number 5?
Pollen
Identify urine artifacts :
What is number 6?
clothing fiber
Identify urine artifacts:
What is number 8?
diaper fiber
Identify urine artifacts :
What is number 9?
cotton fiber