1. What structures are visible in the unstained Tetrahymena sp. ? With iodine? With methyl green ? With nigrosin?

- In an unstained Tetrahymena sp., under the scanner, small circular organisms are seen moving in fast speeds. The cell is stained with Iodine, the nuclei and the glycogen vacuoles are now visible also the cilia of the species is now visible. In Methyl Green Staining, the Micronuclei and Macronuclei also the Nuclei of the cell is now visible with this stain. In Nigrosin it showed a transparent structure but the outline of the species is visible. 2. Do all stains enter the cell or just render more contrast to the background? Stains work in different ways, either they stain only the surface of the cell to make a contrast to to the background while others enter the cell membrane to stain the structures inside the cell. The Iodine and Methyl Green stains are stains that enter the cell and stain specific cell structures while Nigrosin stains the surface of the cell showing the outline of its shape. 3. By providing a dark background, would the cell structures look clearer? - the cell appears as a bright object against a black background. The dark background would show the structures that are hard to locate when using a bright backgound. 4. In your formal report, tabulate the different structures seen under each of the stains used. Explain the mechanism for each reaction.

1. A live, unstained Tetrahymena sp., under the scanner of brightfield microscope are seen as small circular organisms moving in fast speeds. The cell is stained using Lugol's iodine, methyl green, and nigrosin relief method. 2. As mentioned earlier, a brightfield microscope was used to view the prepared slides of Tetrahymena sp. Such microscopes provide a dark specimen viewed over a bright 3. background. This being said, providing a dark background would make the cell structures look clearer. This is because the cell appears as a bright object against a black background. The dark background would show the structures that are hard to locate when using a bright backgound. 4. Lugol's iodine is used to to stain the cilia of the specimen ( Tetrahymena sp.). It stains polysaccharide. In the experiment, nuclei and glyogen vacuoles were also made visible. 5. Methyl green, that is red-brown in physical form, is used to detect phosphates, polyphosphates tocopherol and tocopherol acetate. It stains nucleic acids, chromosomes, brain, spinal cord, nicotine acetylcholine receptor and bacteria. It has a pH range of 0.1 - 2.3. Within the given pH range, methyl green changes color from yellow (0.1) to green (2.3). Methyl green has an absorption of 629 -423 nm. In The experiment, the micronuclei and macronuclei were made visible 6. Nigrosin that is black crystals or powder in physical form, is soluble in water and slightly soluble in ethanol. It is used to detect cells, proteins, keratin fibers, and cell surface. It has an absorption of 570 nm. In the experiment, the specimen showed a transparent structure but the outline of the species is visible. 7. Stains work in different ways, either they stain only the surface of the cell to make a contrast to to the background while others enter the cell membrane to stain the structures inside the cell. The Iodine and Methyl Green are those that enter the cell and stain specific cell structures. Nigrosin, on the other hand, stains the surface of the cell showing the outline of its shape.

Source: 1. "Handbook of Biological dyes and stains : synthesis and industrial applications R. W. Sabnis (2010). John Wiley & Sons, inc., publication" : "http://global-4-lvs-hopper5.opera-mini.net/ha04-02-03/8312/0/-1/chemistry-chemists.com/82746895/handbookof-biological-dyes-and-stains-2010.pdf"