How Gram staining is done?

What are the 5 steps of Gram staining?

What is Gram staining and how does it work?

How does the Gram stain test work?

How do you heat fix a slide?

“Heat-fix” the slide with the specimen by passing it over a heat source, such as a flame, several times using a clothes pin or forceps. The slide should be passed very quickly through the flame and not be heated excessively. Place slide on the staining tray.

How do you Gram stain a plate?

How do you make a Gram stain smear?

How long will you drench the bacterial smear in crystal violet?

Smear and heat fix a clean microscope slide with your bacterial culture. 2. Put 10 to 15 drops of Crystal Violet stain on your bacteria smear and leave on for one minute.

How might culture age affect the results of a spore stain?

Why does culture age affect the results of a spore stain? Older cultures are most likely to produce spores due to depleted nutrition. Younger cultures are most likely to produce spores due to adequate nutrition. Younger cultures are more active metabolically.

How does acid-fast stain work?

The cells in the sample hold onto the dye. The slide is then washed with an acid solution and a different stain is applied. Bacteria that hold onto the first dye are considered “acid-fast” because they resist the acid wash. These types of bacteria are associated with TB and other infections.

What is the safest way of drying bacterial smear and why?

Air-drying ensures that the smear is thin enough to stain. The air-drying step coagulates the proteins in the bacteria. Drying helps remove excess water to ensure optimal heat fixation.

How do you Restain a Gram stain?

The performance of the Gram Stain on any sample requires 4 basic steps that include applying a primary stain (crystal violet) to a heat-fixed smear, followed by the addition of a mordant (Gram’s Iodine), rapid decolorization with alcohol, acetone, or a mixture of alcohol and acetone and lastly, counterstaining with …

How do you make a smear?

Culture from solid media: Using a sterile pipette, add one drop of sterile saline or sterile water to the center of the microscope slide. Aseptically pick a small amount of an isolated colony with a loop and gently mix into the drop of sterile saline or water using circular motions. Mix evenly to make a thin smear.

How do you prepare a microscope slide for bacteria?

Use an inoculating needle to add a drop of saline to a clean slide. Take a sample of a single bacterial colony and mix it into the saline. Let air dry until a white film appears. Heat fix by passing the slide through a flame a few times.

How do you prepare a Gram stain for a smear?

Preparation of the smear is as follows: From broth: Using a cooled, sterile loop, place a loopful of broth on the slide and spread in a circular motion to about 1 cm in diameter. From plated media: Place a drop of sterile water or saline on the slide. Select the isolated colony to be stained.

Why is my Gram stain not working?

Several factors may affect the results of Gram staining: If the smear is too thick, proper decolorizing will not be possible. If the smear is overheated during heat fixing, the cell walls will rupture. Concentration and freshness of reagents may affect the quality of the stain.

How does acid fast stain work?

The cells in the sample hold onto the dye. The slide is then washed with an acid solution and a different stain is applied. Bacteria that hold onto the first dye are considered “acid-fast” because they resist the acid wash. These types of bacteria are associated with TB and other infections.

How do you make a zinc stain?

To prepare Ziehl’s carburized fuchsin working solution, mix 10 ml of solution A with 90 ml of solution B. – Place 300 ml of water in a 1 liter Erlenmeyer flask. Slowly add 100 ml of concentrated sulfuric acid, letting it run down the side of the flask. The mixture will heat up.

How do you do a zinc stain?

How can you mess up a Gram stain?

How will you sterilize a wire loop?

The inoculating loop is sterilised by passing it at an angle through the flame of a gas burner until the entire length of the wire becomes orange from the heat. In this way all contaminants on the wire are incinerated. Never lay the loop down once it is sterilised, or it may again become contaminated.