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Applying a primary stain (crystal violet) to a heat-fixed smear of a bacterial culture. Heat fixation kills some bacteria but is mostly used to affix the bacteria to the slide so that they do not rinse out during the staining procedure. The addition of iodine, which binds to crystal violet and traps it in the cell
Once stained, they do not decolourize. The addition of heat during the staining process is a huge contributing factor. [15] Heat helps open the spore's membrane so the dye can enter. The main purpose of this stain is to show germination of bacterial spores.
Using aseptic technique, prepare and air dried heat fixed slide with the desired organism. Prepare a boiling water bath. Cover the slide with a piece of paper towel and place on staining rack over the water bath. Flood the paper towel on the slide with Malachite Green ( primary stain). Steam the slide for 5 to 7 minutes (mordant).
Major applications include blood smears, bone marrow aspirates, semen analysis and cytology of various body fluids including urine and cerebrospinal fluid. [7] [8] Microbiologic agents, such as bacteria and fungi, also appear more easily in Diff-Quik. [3] This is useful for the detection of for example Helicobacter pylori from gastric and ...
Crystal violet or gentian violet, also known as methyl violet 10B or hexamethyl pararosaniline chloride, is a triarylmethane dye used as a histological stain and in Gram's method of classifying bacteria.
Smears are air dried between 10 and 60 minutes, fixed in 80 vol% ethanol for 5 min at 20-22 °C, rinsed with tap water, and air dried. Films are then immersed in the citrate-phosphate buffer for 5 minutes at 37 °C and gently agitated for about 3 minutes.
Giemsa stain is a classic blood film stain for peripheral blood smears and bone marrow specimens. Erythrocytes stain pink, platelets show a light pale pink, lymphocyte cytoplasm stains sky blue, monocyte cytoplasm stains pale blue, and leukocyte nuclear chromatin stains magenta. It is also used to visualize the classic "safety pin" shape in ...
The cytocentrifugation process can cause cells to appear distorted. Cells located at the centre of the smear may look compressed compared to cells at the periphery. Cell nuclei may develop artifactual clefts, lobes, or holes, [4] and the cytoplasm may appear vacuolated or develop irregular projections. Cytoplasmic granules may be pushed to the ...