A wet mount stain is when a drop of water is placed onto the microscope slide. The water on the slide helps to support the organism and sample. The water fills the space between the cover slip and the slide. This action allows the light from the microscope to pass through the slide and the sample for better visualization of the organisms. A direct stain occurs when a charged color portion of a basic dye like methylene blue combines with the negatively charged portion of and organism allowing the bacterium to become directly stained. In direct staining, the organisms must be fixed by a process such as heat. Fixing the slide prevents the organism form washing off the slide before visualization. This is accomplished by passing a smear of the bacteria through flame. The heat sets the proteins of the organism thus causing the bacteria to attach to the slide. The organism can become damaged from the setting process and the use of heat prior to staining. In indirect staining, the negatively charged colored portion of an acidic dye is repelled by the negatively charged bacterial cell wall. This causes the background to be stained while leaving the organism’s cell wall to remain colorless and unstained.…
What dyes did you used in Gram stain? Crystal violet, gram’s iodine, and safranin violet.…
C: When we are talking about difference between direct and indirect stains, main importance is acidity of the dye. Dye that we are using in direct staining is basic. The colored part of this dye is positive charge, but when in contact with negative cytoplasm of bacteria, at that point bacteria change color and become stained with purple hue. For direct staining we are using crystal violet or methylene blue stain. For indirect staining we are using dyes that are acidic. Usually we are using Congo red, nigrosine or India ink. The acidic dye contains negatively charged color portion. When applied and in contact with…
What is the role of the alcohol wash in the Gram Stain? What color would all bacteria become at the end of the staining procedure if the alcohol step in the gram stain was omitted?…
Gram staining is a technique that was invented by Hans Christian Gram in 1882 (Cantey & Doern, 2015). First, a drop of water was placed on a microscope slide. Using a loop and aseptic techniques (working by an open flame to prevent contamination, and sterilizing the loop in the flame until red hot), a small amount of bacteria from the master plate was mixed with the water then streaked onto the slide. The slide was allowed to dry, then the bacteria was heat fixed onto the slide by running it through the flame twice. A clothespin was attached to the slide and the slide was placed on a staining tray. The smear was covered with crystal violet and allowed to stain the bacteria for two minutes. Then, the slide was rinsed with distilled water at an angle, not directly onto the smear. Grams iodine was added to the slide and allowed to sit for one minute and rinsed again with distilled water. Next, the differentiation step occurred (Bartholomew & Mittwer, 1952) using a small amount of alcohol (about 8 drops) rinsed over the slide, followed immediately by rinsing with distilled water. Safranin was then added to the slide and allowed to sit for 30 seconds then rinsed with distilled water. After blotting the slide with bibulous paper, the slide was viewed under the microscope at 1000X magnification. The counterstain Safranin can be substituted for another…
Simple dyes used to stain bacteria have a positive charge cationic (+), therefore, they are attracted to the bacteria that are usually negatively charged anionic (-)…
This test is performed to get a description of the make-up of the cell membrane of the bacteria. A gram positive bacteria will display a blue/violet pigmentation where a gram negative bacteria will display a pink/red pigmentation. This is due to the outer membrane of gram negative bacteria. My unknown bacteria displays a pink color hinting the presence of a gram negative bacteria.…
Prokaryotes are a large group of organisms with no membrane bound organelles. They consist of two domains: Archaea and Bacteria. These organisms are only found in extreme environments such as volcanoes. Prokaryotes are still being researched and are a very diverse group. In this lab we focused on trying to identify if the bacteria found had a lot of peptidoglycan by gram staining. Testing this could be done by using a Petri dish full of agar and testing different bacteria on it to see if the bacteria obtained is gram positive or gram negative. My hypothesis is there will be a lot of bacterial growth on all of the plate.…
Prepare two slides for gram staining and viewing under a microscope. {Either my gram-stain slides were bad or the microscopes I chose for viewing were not good. No bacteria were found under the microscope.}…
| To determine the ability of a bacterium to fremnet a carbohydrate, make acid or gas…
3. Compare and contrast gram-positive and gram-negative cell walls with regard to (a) sensitivity to antimicrobial agents, (b) resistance to phagocytosis, (c) chemical…
Gram staining showed gram negative rods, a motility test was also conducted to see if the bacterium moved or not, it was found to be none motile. Three different types of agar plates were used, they had two known bacterium put on along with the unknown to be able to compare negative and positive results if the known with the results of the unknown, refer to Barbaro (2016) for how the test were done.…
Secondly both bacteria had to be gram stained, so that they could be identified as gram-positive or as gram-negative. The same gram staining method was used for both bacteria, which consisted of inoculating each bacteria into a…
As I recall from earlier that semester, the first test that needed to be performed was the Gram stain. The Gram stain is a staining technique that aids in distinguishing cell wall characteristics and the cellular morphology. Bacteria will stain Gram positive, which is a purple color, or Gram negative, which is a pink color. Also, the Gram stain can help to identify the shape of the bacterium. The three basic shapes are cocci, bacilli and spirilli, which means that cocci are berry shaped, bacilli are rod shaped and spirilli are spiral shaped. My unknown bacteria stained purple which means that it is Gram-positive and was berry shaped meaning that the bacterium is cocci. These results have led me to the next step for me to master, the catalase test.…
Stains are solutions that contain a solute called a chromophore dissolved in a solvent. A chromophore is the color possessing portion of the solution and is therefore responsible for the stains color. Bacterial cells usually have a negative surface charge, meaning that a positively charged stain is needed to stain the surface of the cell. Most stains are basic and have a positively charged chromophore. When the stain is applied, there is an attraction between the negatively charged cell surface and the positively charged chromophore, leading to the surface…