Part 3: preparation of the smear bacterial suspensions in broth: with a sterile cooled loop, place a loopful of the broth culture on the slide spread by means of circular motion of the inoculating loop to about one centimeter in diameter excessive spreading may result in disruption of cellular arrangement. Bacterial smear is defined as a dehydrated or dried preparation of a bacterial suspension (cells) on a clean glass slide the source of bacteria for the preparation or making of a smear can be from an agar slant, broth (liquid) culture and from a culture plate. A bacterial smear on a slide is normally made every time before one stains bacteria for microscopic observation besides placing bacteria on a slide, this procedure dries and fixes the bacteria to the glass, so that the cells are less likely to be washed away during the staining procedure. Do not use forced heat drying forced drying will distort bacterial cells and other materials 4 hold the smear with forceps and fix the smear by passing it through a flame (smear side up) three or four times. Describe the methods of preparation of a bacterial smear from broth and solid media 2 describe the attributes of an ideal smear 3 identify the factors which can result in a less-than-ideal smear 4 define the purpose of the specific staining methods used to stain bacteria.
Heat fixing bacterial smears kills the bacterial cells so that they are fixed in place and ready for staining heat fixing also adheres the bacterial cells to the slide and allows the sample to take up the stain more easily. In order to heat fix a bacterial smear, it is necessary to first let the bacterial sample air dry then either place the slide in the slide holder of a microincinerator, or pass the dried slide through the flame of a bunsen burner 3 or 4 times, smear side facing uponce the slide is heat fixed, it can then be stained. Steps of microbial smear preparation : 1handle a clean slide by its edge, label the target place at the bottom side o the 3if the bacterial culture was broth, shake the culture and transfer loopful of broth to the center of the slide and spread over the target circle. Smear preparation the first step in most bacterial staining procedures is the preparation of a smear in a smear preparation, cells from a culture are spread in a thin film over a small area of a microscope slide, dried, and then fixed to the slide by heating or other chemical fixatives.
Edit article how to make a sterile bacterial smear four parts: cleaning your work space making yourself sterile keeping your culture pure cleaning up community q&a when making a bacterial smear, it is important to keep cultures and slides uncontaminated to properly view the microbe under a microscope. Procedure simple staining procedure preparation of a smear and heat fixing using a sterilized inoculating loop, transfer loopful of liquid suspension containing bacteria to a slide (clean grease free microscopic slide) or transfer an isolated colony from a culture plate to a slide with a water drop. Because bacteria are, for the most part, transparent, we use stains to give them color for microscopic observation making a bacterial smear prepares the bacteria to be stained and is the first step of most staining procedures. Slide/bacterial sample preparation and then on to a simple staining method for the visualization of the bacteria (remember that bacteria are very difficult to see without stains) materials and methods materials preparing a bacterial smear on microscope slides: author.
A bacterial smear is a dried preparation of bacterial cells on a glass slide smears may be made from a dry culture (from a slant, stab or plate), or from a wet/liquid culture in preparing bacterial smears , the following qualities must be observed: 1. Staining: bacterial smear preparation before a specimen can be observed it must be prepared a smear is a thin layer of cells on a slide a bacterial smear can be made from a liquid culture, or a culture on solid media. Since staining of bacterial cells is relatively fast, inexpensive, and simple, it is the most commonly used technique to visualize bacterial cells staining not only makes bacteria more easily seen, but it allows their morphology (eg size and shape) to be visualized more easily.
Smear preparation total cards 18 subject microbiology level undergraduate 2 created 10/23/2011 how does one prepare a smear if the bacteria grow in solid medium definition place 1 or 2 loopfuls of water on the slide and then use an inoculating loop to disperse the organism in the water. Bacterial smear and staining preparation smear procedure: with the pen (permanent ink)/pencil, mark the name of the bacterial culture in the far left corner on each slides. Smear preparation technique consists of spreading small volume of sample on a slide and air drying the film before staining and microscopy bacterial smears must be prepared prior to any of the staining techniques. Preparing a bacterial smear slide preparing a bacterial smear slide is useful for microscopic examination of dead bacterial cells 1 place a very small loopful of deionized water on the slide (you can use the deionized water bottle on your bench-remove the cover and dip your loop in since sterility is not required for this step.
In most microbiological staining procedures, the bacteria are first fixed to the slide by the heat fixed smear preparation of the heat fixed smear after preparation of the heat fixed smear, the cells may then be stained the stains are smear the bacteria on the end of the loop into one of the drops of water on the slide. 1 preparing a bacterial smear -since bacterial cells are so small, special problems arise in the preparation of a good smear -there are 3 principal precautions that you must take to avoid problems. Created date: 9/16/2005 1:11:26 pm. The smear layer is a layer of microcrystalline and organic particle debris that is found spread on root canal walls after root canal instrumentation it was first described in 1975 and much research has been performed since then to evaluate its importance in assisting or preventing the penetration of bacteria into the dentinal tubules.
Choose an area of the smear in which the cells are well spread in a monolayer center the area to be studied, apply oil directly to the smear, and focus the smear under oil with the 100x objective. A bacterial smear on a slide is normally made every time one stains bacteria for microscopic observation besides placing bacteria on a slide, this procedure dries and fixes the bacteria to the glass, so that the cells are not washed away during the staining procedure. A bacterial smear is required particularly for the microscopic examination of the specimen or the culture for the identification of bacteria which is done by preparing the smear from the specimen or culture followed by staining with the appropriate stains, eg gram staining the purpose of making the smear is to fix the specimen or the.