Describe how to make a wet mount slide?

Answer 1

Proper specimen collection and handling of vaginal samples is crucial for accurate results. The vaginal vault and walls should be swabbed using one or two swabs. If any fluid has pooled in other areas, these areas should be swabbed as well. The swabs should then be placed in a tube containing 0.5 ml saline and examined within two hours of collection. The sample should remain at room temperature.

After obtaining the sample, it is equally important to properly prepare the slide for examination. The direct, unstained slide should be prepared as follows:

1. Vigorously mix the swab(s) in and out of the saline making sure to collect all the material adhering to the side of the tube.
2. Remove the swab from the saline and depress onto a clean, dry microscope slide expressing a small amount of fluid.
3. Coverslip the sample and examine under a microscope.

The KOH slide may be prepared by adding a drop of KOH to the sample after following the directions as noted above. The saline slide should be examined first to allow the KOH to properly digest other cellular elements in the sample such as epithelial and blood cells.

The pH and the amine ("whiff") test often are performed before the microscopic examination. The clinician tests the pH at the time of sample collection. A pH greater than 4.5 may indicate bacterial vaginosis (BV) or trichomoniasis. The amine test is performed prior to cover slipping the KOH preparation. A "fishy" or amine odor is characteristic in the presence of Trichomonas vaginalis and Gardnerella vaginalis.

Begin the microscopic review of the slide by examining the saline preparation. The examination should begin using the 10x objective noting cellular distribution and obvious cellular and fungal elements. The 40x objective is used to identify the presence of white and red blood cells, quantity and type of bacteria present, clue cells, motile Trichomonas, yeasts, and fungal hyphae. The KOH slide should be examined with the 10x objective for any yeast and hyphae present and the 40x objective is used to distinguish smaller budding yeasts and hyphae.

Common causes of bacterial vaginitis include Trichomonas vaginalis, Gardnerella vaginalis, and Candida albicans. All of these infections can be diagnosed by direct examination. The absence or presence of white blood cells and normal vaginal flora are clues that can be directly related to the causative agent of infection. The amine test and vaginal pH also can help to determine the cause of infection.

Trichomoniasis is caused by Trichomonas vaginalis. This single-celled parasite is transmitted sexually. Commonly, the parasite is motile with its flagella whipping back and forth. However, it may not be motile and can easily be missed or confused with white blood cells. The infection is associated with large numbers of white cells with a positive amine test and the absence of normal vaginal flora.

Gardnerella vaginalis is often characterized as non-specific vaginitis. Microscopically it is characterized by a lack of normal vaginal flora and a predominance of many small coccobacilli. The small bacteria adhere to the surface of the epithelial cells creating a speckled appearance. These speckled cells are called "clue cells" and their presence is considered diagnostic for Gardnerella vaginalis induced vaginitis. Gardnerella infections do not provoke a large white blood cell response and have a positive amine test.

Yeast infections, commonly caused by Candida albicans, are easily identified upon direct examination. As expected, the direct examination will yield budding yeasts and hyphae. The use of KOH enhances the recovery of these fungal elements. Yeasts provoke a large white blood cell response with a negative amine test. Normal vaginal flora will be present.

Laboratories should establish their own criteria for diagnosing vaginitis. In most cases, the direct examination of vaginal secretions in a symptomatic patient will result in the proper diagnosis of the infection and allow for prompt treatment. Use of vaginal lubricants, douches, tampons, contraceptive jelly, and medications may interfere with the examination. Patients should avoid using these items before sample collection. In some cases, the Gram stain may be used to aid in the diagnosis of these patients.

Answer 2

Living specimens do not survive long in the heat from an intense microscope illuminator bulb, usually because the specimen dries up. This problem is easily solved by preparing a vaseline chamber. Simply take a single cover slip and hold it between thumb and forefinger by the edges. Pick up some vaseline on the other forefinger and rub it over your thumb to make a film. Scrape your thumb carefully on each edge of the coverslip to make a continuous vaseline ledge. Place a drop or two of suspension on a clean slide, and turn the coverslip over on top of the drop. Press down the edges to seal the chamber against evaporation. When preparing a vaseline mount, keep in mind that the image becomes degraded with thicker mounts, especially at high powers in dark field or phase contrast. Unless the specimen is large and fragile enough to be damaged by pressing down too hard on the coverslip, keep the chamber depth very shallow.

Answer 3

obtain a clean microscope slide and cover slip. in a wet mount, the coverslip serves three functions: it flattens the sample, it prevents from drying out, and it protects the objective lens from contaminations. place your sample in the centre of the slide. the specimen must be thin for light to pass through. with an eyedropper, place drop of water on the sample. place the coverslip at an angle at one end of the drop of water. carefully lower the coverslp to cover the sample, being careful not to trap any air. it may be helpful to use a probe ot toothpick to lower the coverslip. if you do get air bubbles, gently tap the slide with a probe to release them. well this is more like how to prepare..hope it make sense guys!(p.s. i actually hate Biology but i like working on microscopes:))

Answer 4

Preparing a microscope slide that contains an insect wing can be done in a couple of steps. Make sure the two slides that you will use are clean. Place the insect wing in between the two slides. Place the slide under the microscope.

Answer 5

1. Get a Clean slide,Put 1 drop of water on the slide 2. put your speciem on the clean slide 3. put the Cover slip over your specim 4. look under microcope!

Answer 6

you put vaseline and suspension on it and plop the vermin on it!