Lab 2 Staining Methods Reading PDF

Part 3 Staining Methods Exercises 5 Preparation of Smears and Simple Staining 6 Negative Staining 7 Gram Staining 8 Acid-Fast Staining 9 Structural Stains (Endospore, Capsule, and Flagella) 10 Morphologic Unknown ASM: Demonstrate ability in properly preparing slides for microbiological exami...

Part 3 Staining Methods Exercises 5 Preparation of Smears and Simple Staining 6 Negative Staining 7 Gram Staining 8 Acid-Fast Staining 9 Structural Stains (Endospore, Capsule, and Flagella) 10 Morphologic Unknown ASM: Demonstrate ability in properly preparing slides for microbiological examination including sterilizing and maintaining sterility of transfer instruments and recording microscopic observations. In 1877, Robert Koch wrote, How many incomplete and false observations might have remained unpublished instead of swelling the bacterial literature into a turbid stream, if investigators had checked their preparations with each other?\* \*Quoted in H. A. Lechevalier and M. Solotorovsky. Three Centuries of Microbiology. New York: Dover Publications, 1974, p. 79. To solve this problem, he introduced into microbiology the procedures of air-drying, chemical fixation, and staining with aniline dyes. In addition to making a lasting preparation, staining bacteria enhances the contrast between bacteria and the surrounding material and permits observation of greater detail and resolution than do wet-mount procedures. Microorganisms are prepared for staining by smearing them onto a microscope slide. In these exercises, bacteria will be transferred from growth or culture media to microscope slides using an inoculating loop. Staining techniques may involve simple stains, in which only one reagent is used and all bacteria are usually stained similarly (Exercises 5 and 6), or differential stains, in which multiple reagents are used and bacteria react to the reagents differently (Exercises 7 and 8). Structural stains are used to identify specific parts of microorganisms (Exercise 9). Most of the bacteria that are grown in laboratories are rod shaped bacilli or cocci. Rods and cocci will be used throughout this part. Staining and microscopic examination are usually the first steps in identifying microorganisms. In Exercise 10, Morphologic Unknown, you will be asked to characterize selected bacteria by their microscopic appearance. Case Study: A Summer Rash! Peter and Mary live in Minneapolis, Minnesota, with their two active sons. Peter is a microbiologist at General Mills and loves his job. Mary is a teacher at a neighborhood elementary school. It had been a hot summer, and the kids were ready for bed after playing outside all day. Five-year-old son Shawn had been scratching his skin, and when Peter looked closely, he saw a small, pimple-like rash over Shawn's chest and arms. He quickly examined his three-year-old son and, to his dismay, found a similar rash on Todd's abdomen. He immediately started to worry about viral diseases (such as measles) and bacterial diseases (impetigo, erysipelas, or even necrotizing fasciitis). Mary quickly reported that she had seen a similar rash in her third-grade class and suggested that they call their neighbors, the Fredricksons, whose children had been playing with Shawn and Todd. The Fredricksons examined their children and were surprised to discover that one of them had a light rash. All three children were active with no apparent pain and did not have an elevated temperature. Mary, the voice of reason, suggested that they call the Urgent Care hotline before rushing off to an expensive visit to the Emergency Room. Sally, the nurse practitioner who answered their call, asked Mary several questions about the rash and the children's activities. It was the middle of July, and because of the hot weather, the children had been spending many afternoons playing in the backyard plastic pool. Peter had emptied the pool each morning and filled it with fresh water, which heated up nicely by the afternoon. Sally suspected that the three children had dermatitis from their repeated use of the pool, which never really dried out and never was disinfected. Sally did not recommend the use of antibiotics but advised the parents simply to watch the rash and to minimize the children's scratching. She indicated that the rash would go away in about a week and said they did not need to come in. Peter, ever the microbiologist, took several swab cultures of the pool, selecting areas by the seams and slimy areas that appeared to be a biofilm. He took the swabs to work and cultured them on nutrient agar. After 24 hours, bacteria had grown on the plates and produced a bluish-green, water-soluble pigment. The culture was Gram stained, and a gram-negative, very small bacillus was visible. The children's rash lasted several days but then gradually went away without complications. Questions Why did the Gram stain results rule out impetigo, erysipelas, and necrotizing fasciitis? Give a staining technique that would help determine whether the culprit was a Bacillus. How did the bacteria enter the skin? Part 3 Staining Methods Exercises 5 Preparation of Smears and Simple Staining 6 Negative Staining 7 Gram Staining 8 Acid-Fast Staining 9 Structural Stains (Endospore, Capsule, and Flagella) 10 Morphologic Unknown ASM: Demonstrate ability in properly preparing slides for microbiological examination including sterilizing and maintaining sterility of transfer instruments and recording microscopic observations. In 1877, Robert Koch wrote, How many incomplete and false observations might have remained unpublished instead of swelling the bacterial literature into a turbid stream, if investigators had checked their preparations with each other?\* \*Quoted in H. A. Lechevalier and M. Solotorovsky. Three Centuries of Microbiology. New York: Dover Publications, 1974, p. 79. To solve this problem, he introduced into microbiology the procedures of air-drying, chemical fixation, and staining with aniline dyes. In addition to making a lasting preparation, staining bacteria enhances the contrast between bacteria and the surrounding material and permits observation of greater detail and resolution than do wet-mount procedures. Microorganisms are prepared for staining by smearing them onto a microscope slide. In these exercises, bacteria will be transferred from growth or culture media to microscope slides using an inoculating loop. Staining techniques may involve simple stains, in which only one reagent is used and all bacteria are usually stained similarly (Exercises 5 and 6), or differential stains, in which multiple reagents are used and bacteria react to the reagents differently (Exercises 7 and 8). Structural stains are used to identify specific parts of microorganisms (Exercise 9). Most of the bacteria that are grown in laboratories are rod shaped bacilli or cocci. Rods and cocci will be used throughout this part. Staining and microscopic examination are usually the first steps in identifying microorganisms. In Exercise 10, Morphologic Unknown, you will be asked to characterize selected bacteria by their microscopic appearance. Case Study: A Summer Rash! Peter and Mary live in Minneapolis, Minnesota, with their two active sons. Peter is a microbiologist at General Mills and loves his job. Mary is a teacher at a neighborhood elementary school. It had been a hot summer, and the kids were ready for bed after playing outside all day. Five-year-old son Shawn had been scratching his skin, and when Peter looked closely, he saw a small, pimple-like rash over Shawn's chest and arms. He quickly examined his three-year-old son and, to his dismay, found a similar rash on Todd's abdomen. He immediately started to worry about viral diseases (such as measles) and bacterial diseases (impetigo, erysipelas, or even necrotizing fasciitis). Mary quickly reported that she had seen a similar rash in her third-grade class and suggested that they call their neighbors, the Fredricksons, whose children had been playing with Shawn and Todd. The Fredricksons examined their children and were surprised to discover that one of them had a light rash. All three children were active with no apparent pain and did not have an elevated temperature. Mary, the voice of reason, suggested that they call the Urgent Care hotline before rushing off to an expensive visit to the Emergency Room. Sally, the nurse practitioner who answered their call, asked Mary several questions about the rash and the children's activities. It was the middle of July, and because of the hot weather, the children had been spending many afternoons playing in the backyard plastic pool. Peter had emptied the pool each morning and filled it with fresh water, which heated up nicely by the afternoon. Sally suspected that the three children had dermatitis from their repeated use of the pool, which never really dried out and never was disinfected. Sally did not recommend the use of antibiotics but advised the parents simply to watch the rash and to minimize the children's scratching. She indicated that the rash would go away in about a week and said they did not need to come in. Peter, ever the microbiologist, took several swab cultures of the pool, selecting areas by the seams and slimy areas that appeared to be a biofilm. He took the swabs to work and cultured them on nutrient agar. After 24 hours, bacteria had grown on the plates and produced a bluish-green, water-soluble pigment. The culture was Gram stained, and a gram-negative, very small bacillus was visible. The children's rash lasted several days but then gradually went away without complications. Questions Why did the Gram stain results rule out impetigo, erysipelas, and necrotizing fasciitis? Give a staining technique that would help determine whether the culprit was a Bacillus. How did the bacteria enter the skin? Part 3 Staining Methods Exercises 5 Preparation of Smears and Simple Staining 6 Negative Staining 7 Gram Staining 8 Acid-Fast Staining 9 Structural Stains (Endospore, Capsule, and Flagella) 10 Morphologic Unknown ASM: Demonstrate ability in properly preparing slides for microbiological examination including sterilizing and maintaining sterility of transfer instruments and recording microscopic observations. In 1877, Robert Koch wrote, How many incomplete and false observations might have remained unpublished instead of swelling the bacterial literature into a turbid stream, if investigators had checked their preparations with each other?\* \*Quoted in H. A. Lechevalier and M. Solotorovsky. Three Centuries of Microbiology. New York: Dover Publications, 1974, p. 79. To solve this problem, he introduced into microbiology the procedures of air-drying, chemical fixation, and staining with aniline dyes. In addition to making a lasting preparation, staining bacteria enhances the contrast between bacteria and the surrounding material and permits observation of greater detail and resolution than do wet-mount procedures. Microorganisms are prepared for staining by smearing them onto a microscope slide. In these exercises, bacteria will be transferred from growth or culture media to microscope slides using an inoculating loop. Staining techniques may involve simple stains, in which only one reagent is used and all bacteria are usually stained similarly (Exercises 5 and 6), or differential stains, in which multiple reagents are used and bacteria react to the reagents differently (Exercises 7 and 8). Structural stains are used to identify specific parts of microorganisms (Exercise 9). Most of the bacteria that are grown in laboratories are rod shaped bacilli or cocci. Rods and cocci will be used throughout this part. Staining and microscopic examination are usually the first steps in identifying microorganisms. In Exercise 10, Morphologic Unknown, you will be asked to characterize selected bacteria by their microscopic appearance. Case Study: A Summer Rash! Peter and Mary live in Minneapolis, Minnesota, with their two active sons. Peter is a microbiologist at General Mills and loves his job. Mary is a teacher at a neighborhood elementary school. It had been a hot summer, and the kids were ready for bed after playing outside all day. Five-year-old son Shawn had been scratching his skin, and when Peter looked closely, he saw a small, pimple-like rash over Shawn's chest and arms. He quickly examined his three-year-old son and, to his dismay, found a similar rash on Todd's abdomen. He immediately started to worry about viral diseases (such as measles) and bacterial diseases (impetigo, erysipelas, or even necrotizing fasciitis). Mary quickly reported that she had seen a similar rash in her third-grade class and suggested that they call their neighbors, the Fredricksons, whose children had been playing with Shawn and Todd. The Fredricksons examined their children and were surprised to discover that one of them had a light rash. All three children were active with no apparent pain and did not have an elevated temperature. Mary, the voice of reason, suggested that they call the Urgent Care hotline before rushing off to an expensive visit to the Emergency Room. Sally, the nurse practitioner who answered their call, asked Mary several questions about the rash and the children's activities. It was the middle of July, and because of the hot weather, the children had been spending many afternoons playing in the backyard plastic pool. Peter had emptied the pool each morning and filled it with fresh water, which heated up nicely by the afternoon. Sally suspected that the three children had dermatitis from their repeated use of the pool, which never really dried out and never was disinfected. Sally did not recommend the use of antibiotics but advised the parents simply to watch the rash and to minimize the children's scratching. She indicated that the rash would go away in about a week and said they did not need to come in. Peter, ever the microbiologist, took several swab cultures of the pool, selecting areas by the seams and slimy areas that appeared to be a biofilm. He took the swabs to work and cultured them on nutrient agar. After 24 hours, bacteria had grown on the plates and produced a bluish-green, water-soluble pigment. The culture was Gram stained, and a gram-negative, very small bacillus was visible. The children's rash lasted several days but then gradually went away without complications. Questions Why did the Gram stain results rule out impetigo, erysipelas, and necrotizing fasciitis? Give a staining technique that would help determine whether the culprit was a Bacillus. How did the bacteria enter the skin? Part 3 Staining Methods Exercises 5 Preparation of Smears and Simple Staining 6 Negative Staining 7 Gram Staining 8 Acid-Fast Staining 9 Structural Stains (Endospore, Capsule, and Flagella) 10 Morphologic Unknown ASM: Demonstrate ability in properly preparing slides for microbiological examination including sterilizing and maintaining sterility of transfer instruments and recording microscopic observations. In 1877, Robert Koch wrote, How many incomplete and false observations might have remained unpublished instead of swelling the bacterial literature into a turbid stream, if investigators had checked their preparations with each other?\* \*Quoted in H. A. Lechevalier and M. Solotorovsky. Three Centuries of Microbiology. New York: Dover Publications, 1974, p. 79. To solve this problem, he introduced into microbiology the procedures of air-drying, chemical fixation, and staining with aniline dyes. In addition to making a lasting preparation, staining bacteria enhances the contrast between bacteria and the surrounding material and permits observation of greater detail and resolution than do wet-mount procedures. Microorganisms are prepared for staining by smearing them onto a microscope slide. In these exercises, bacteria will be transferred from growth or culture media to microscope slides using an inoculating loop. Staining techniques may involve simple stains, in which only one reagent is used and all bacteria are usually stained similarly (Exercises 5 and 6), or differential stains, in which multiple reagents are used and bacteria react to the reagents differently (Exercises 7 and 8). Structural stains are used to identify specific parts of microorganisms (Exercise 9). Most of the bacteria that are grown in laboratories are rod shaped bacilli or cocci. Rods and cocci will be used throughout this part. Staining and microscopic examination are usually the first steps in identifying microorganisms. In Exercise 10, Morphologic Unknown, you will be asked to characterize selected bacteria by their microscopic appearance. Case Study: A Summer Rash! Peter and Mary live in Minneapolis, Minnesota, with their two active sons. Peter is a microbiologist at General Mills and loves his job. Mary is a teacher at a neighborhood elementary school. It had been a hot summer, and the kids were ready for bed after playing outside all day. Five-year-old son Shawn had been scratching his skin, and when Peter looked closely, he saw a small, pimple-like rash over Shawn's chest and arms. He quickly examined his three-year-old son and, to his dismay, found a similar rash on Todd's abdomen. He immediately started to worry about viral diseases (such as measles) and bacterial diseases (impetigo, erysipelas, or even necrotizing fasciitis). Mary quickly reported that she had seen a similar rash in her third-grade class and suggested that they call their neighbors, the Fredricksons, whose children had been playing with Shawn and Todd. The Fredricksons examined their children and were surprised to discover that one of them had a light rash. All three children were active with no apparent pain and did not have an elevated temperature. Mary, the voice of reason, suggested that they call the Urgent Care hotline before rushing off to an expensive visit to the Emergency Room. Sally, the nurse practitioner who answered their call, asked Mary several questions about the rash and the children's activities. It was the middle of July, and because of the hot weather, the children had been spending many afternoons playing in the backyard plastic pool. Peter had emptied the pool each morning and filled it with fresh water, which heated up nicely by the afternoon. Sally suspected that the three children had dermatitis from their repeated use of the pool, which never really dried out and never was disinfected. Sally did not recommend the use of antibiotics but advised the parents simply to watch the rash and to minimize the children's scratching. She indicated that the rash would go away in about a week and said they did not need to come in. Peter, ever the microbiologist, took several swab cultures of the pool, selecting areas by the seams and slimy areas that appeared to be a biofilm. He took the swabs to work and cultured them on nutrient agar. After 24 hours, bacteria had grown on the plates and produced a bluish-green, water-soluble pigment. The culture was Gram stained, and a gram-negative, very small bacillus was visible. The children's rash lasted several days but then gradually went away without complications. Questions Why did the Gram stain results rule out impetigo, erysipelas, and necrotizing fasciitis? Give a staining technique that would help determine whether the culprit was a Bacillus. How did the bacteria enter the skin? Part 3 Staining Methods Exercises 5 Preparation of Smears and Simple Staining 6 Negative Staining 7 Gram Staining 8 Acid-Fast Staining 9 Structural Stains (Endospore, Capsule, and Flagella) 10 Morphologic Unknown ASM: Demonstrate ability in properly preparing slides for microbiological examination including sterilizing and maintaining sterility of transfer instruments and recording microscopic observations. In 1877, Robert Koch wrote, How many incomplete and false observations might have remained unpublished instead of swelling the bacterial literature into a turbid stream, if investigators had checked their preparations with each other?\* \*Quoted in H. A. Lechevalier and M. Solotorovsky. Three Centuries of Microbiology. New York: Dover Publications, 1974, p. 79. To solve this problem, he introduced into microbiology the procedures of air-drying, chemical fixation, and staining with aniline dyes. In addition to making a lasting preparation, staining bacteria enhances the contrast between bacteria and the surrounding material and permits observation of greater detail and resolution than do wet-mount procedures. Microorganisms are prepared for staining by smearing them onto a microscope slide. In these exercises, bacteria will be transferred from growth or culture media to microscope slides using an inoculating loop. Staining techniques may involve simple stains, in which only one reagent is used and all bacteria are usually stained similarly (Exercises 5 and 6), or differential stains, in which multiple reagents are used and bacteria react to the reagents differently (Exercises 7 and 8). Structural stains are used to identify specific parts of microorganisms (Exercise 9). Most of the bacteria that are grown in laboratories are rod shaped bacilli or cocci. Rods and cocci will be used throughout this part. Staining and microscopic examination are usually the first steps in identifying microorganisms. In Exercise 10, Morphologic Unknown, you will be asked to characterize selected bacteria by their microscopic appearance. Case Study: A Summer Rash! Peter and Mary live in Minneapolis, Minnesota, with their two active sons. Peter is a microbiologist at General Mills and loves his job. Mary is a teacher at a neighborhood elementary school. It had been a hot summer, and the kids were ready for bed after playing outside all day. Five-year-old son Shawn had been scratching his skin, and when Peter looked closely, he saw a small, pimple-like rash over Shawn's chest and arms. He quickly examined his three-year-old son and, to his dismay, found a similar rash on Todd's abdomen. He immediately started to worry about viral diseases (such as measles) and bacterial diseases (impetigo, erysipelas, or even necrotizing fasciitis). Mary quickly reported that she had seen a similar rash in her third-grade class and suggested that they call their neighbors, the Fredricksons, whose children had been playing with Shawn and Todd. The Fredricksons examined their children and were surprised to discover that one of them had a light rash. All three children were active with no apparent pain and did not have an elevated temperature. Mary, the voice of reason, suggested that they call the Urgent Care hotline before rushing off to an expensive visit to the Emergency Room. Sally, the nurse practitioner who answered their call, asked Mary several questions about the rash and the children's activities. It was the middle of July, and because of the hot weather, the children had been spending many afternoons playing in the backyard plastic pool. Peter had emptied the pool each morning and filled it with fresh water, which heated up nicely by the afternoon. Sally suspected that the three children had dermatitis from their repeated use of the pool, which never really dried out and never was disinfected. Sally did not recommend the use of antibiotics but advised the parents simply to watch the rash and to minimize the children's scratching. She indicated that the rash would go away in about a week and said they did not need to come in. Peter, ever the microbiologist, took several swab cultures of the pool, selecting areas by the seams and slimy areas that appeared to be a biofilm. He took the swabs to work and cultured them on nutrient agar. After 24 hours, bacteria had grown on the plates and produced a bluish-green, water-soluble pigment. The culture was Gram stained, and a gram-negative, very small bacillus was visible. The children's rash lasted several days but then gradually went away without complications. Questions Why did the Gram stain results rule out impetigo, erysipelas, and necrotizing fasciitis? Give a staining technique that would help determine whether the culprit was a Bacillus. How did the bacteria enter the skin? Part 3 Staining Methods Exercises 5 Preparation of Smears and Simple Staining 6 Negative Staining 7 Gram Staining 8 Acid-Fast Staining 9 Structural Stains (Endospore, Capsule, and Flagella) 10 Morphologic Unknown ASM: Demonstrate ability in properly preparing slides for microbiological examination including sterilizing and maintaining sterility of transfer instruments and recording microscopic observations. In 1877, Robert Koch wrote, How many incomplete and false observations might have remained unpublished instead of swelling the bacterial literature into a turbid stream, if investigators had checked their preparations with each other?\* \*Quoted in H. A. Lechevalier and M. Solotorovsky. Three Centuries of Microbiology. New York: Dover Publications, 1974, p. 79. To solve this problem, he introduced into microbiology the procedures of air-drying, chemical fixation, and staining with aniline dyes. In addition to making a lasting preparation, staining bacteria enhances the contrast between bacteria and the surrounding material and permits observation of greater detail and resolution than do wet-mount procedures. Microorganisms are prepared for staining by smearing them onto a microscope slide. In these exercises, bacteria will be transferred from growth or culture media to microscope slides using an inoculating loop. Staining techniques may involve simple stains, in which only one reagent is used and all bacteria are usually stained similarly (Exercises 5 and 6), or differential stains, in which multiple reagents are used and bacteria react to the reagents differently (Exercises 7 and 8). Structural stains are used to identify specific parts of microorganisms (Exercise 9). Most of the bacteria that are grown in laboratories are rod shaped bacilli or cocci. Rods and cocci will be used throughout this part. Staining and microscopic examination are usually the first steps in identifying microorganisms. In Exercise 10, Morphologic Unknown, you will be asked to characterize selected bacteria by their microscopic appearance. Case Study: A Summer Rash! Peter and Mary live in Minneapolis, Minnesota, with their two active sons. Peter is a microbiologist at General Mills and loves his job. Mary is a teacher at a neighborhood elementary school. It had been a hot summer, and the kids were ready for bed after playing outside all day. Five-year-old son Shawn had been scratching his skin, and when Peter looked closely, he saw a small, pimple-like rash over Shawn's chest and arms. He quickly examined his three-year-old son and, to his dismay, found a similar rash on Todd's abdomen. He immediately started to worry about viral diseases (such as measles) and bacterial diseases (impetigo, erysipelas, or even necrotizing fasciitis). Mary quickly reported that she had seen a similar rash in her third-grade class and suggested that they call their neighbors, the Fredricksons, whose children had been playing with Shawn and Todd. The Fredricksons examined their children and were surprised to discover that one of them had a light rash. All three children were active with no apparent pain and did not have an elevated temperature. Mary, the voice of reason, suggested that they call the Urgent Care hotline before rushing off to an expensive visit to the Emergency Room. Sally, the nurse practitioner who answered their call, asked Mary several questions about the rash and the children's activities. It was the middle of July, and because of the hot weather, the children had been spending many afternoons playing in the backyard plastic pool. Peter had emptied the pool each morning and filled it with fresh water, which heated up nicely by the afternoon. Sally suspected that the three children had dermatitis from their repeated use of the pool, which never really dried out and never was disinfected. Sally did not recommend the use of antibiotics but advised the parents simply to watch the rash and to minimize the children's scratching. She indicated that the rash would go away in about a week and said they did not need to come in. Peter, ever the microbiologist, took several swab cultures of the pool, selecting areas by the seams and slimy areas that appeared to be a biofilm. He took the swabs to work and cultured them on nutrient agar. After 24 hours, bacteria had grown on the plates and produced a bluish-green, water-soluble pigment. The culture was Gram stained, and a gram-negative, very small bacillus was visible. The children's rash lasted several days but then gradually went away without complications. Questions Why did the Gram stain results rule out impetigo, erysipelas, and necrotizing fasciitis? Give a staining technique that would help determine whether the culprit was a Bacillus. How did the bacteria enter the skin?