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Microbiology CH 3

by: Evelyn Sanchez

Microbiology CH 3 2440

Evelyn Sanchez
Texas State

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Here are some notes over microscopy in chapter 3.
principles of Microbiology
Class Notes
Microbiology, Microscopy, Microscopes
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This 7 page Class Notes was uploaded by Evelyn Sanchez on Friday September 9, 2016. The Class Notes belongs to 2440 at Texas State University taught by Abel in Fall 2016. Since its upload, it has received 6 views. For similar materials see principles of Microbiology in Microbiology at Texas State University.


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Date Created: 09/09/16
Microorganism­ Must be observed through a microscope. ­Many microbes must undergo staining procedures before their cell walls, capsules, etc. lose  their colorless state. ­Microorganisms are measured in small units, such as micrometers or nanometers. Microscopy Light microscopy – Refers to the use of any kind of microscope that uses visible light to observe  specimens. (Any microscope that uses light)  ­Compound light microscope (LM) : Has a series of lenses, and uses visible light as a  source. The series of lenses forms a clear focused image. This is achieved when light from an  illuminator (the light source) passes through a condenser. Then the light goes through the  specimen into the objective lenses. The image is then magnified by the ocular lenses (the  eyepiece). Total Magnification = Objective lens magnification x Ocular lens magnification Resolution­ The ability of lenses to distinguish fine detail and structure. Refractive index­ A measure of the light­bending ability of a medium.  Darkfeild microscopy : Used to examine live microorganisms that either are invisible in  light, or cannot be stained. Uses a darkfield condenser that contains an opaque disc. Disc blocks  light that would normally enter objective lens. Because there is no direct background light, the  specimen appears light against a black background.  Phase­Contrast microscopy : Useful because the internal structures of a cell become  more defined. Has two sets of light rays that increases the contrast.  Differential interference contrast microscopy (DIC) : Similar to phase­contrast, but splits  up wavelength of light into different colors with a prism. Florescence microscopy : Uses ultraviolet rays, and gives off light at longer wavelengths  making it more visible. Specimens are stained with fluorochromes which makes them appear  luminescent under UV light.  Confocal microscopy : Used to reconstruct 3D images. Like florescence they are also dyed with  fluorochromes, but instead of illuminating the entire specimen, only one plane of a small region  is illuminated with short wavelengths. Basically cuts the image into tiny sections and then put it  back together in a 3D picture.  Two photon microscopy : Has more resolution than confocal and uses long wavelength light  instead of one.  Scanning acoustic microscopy : Interprets a sound wave sent through specimens to create a 3D  picture. Has a resolution of about 1 micrometer.  Electron Microscopy – Beams of electrons are used instead of light. Has the highest resolution  compared to other microscopes. Doesn’t contain ocular lenses so you don’t look directly at a  light beam.  Transmission electron microscopy : Has a resolution of 10 picometers. Beam of electrons passes through an ultrathin section of specimen.  Scanning electron microscopy : Provides striking 3D images of specimens.  Atomic forces microscopy – A metal/diamond probes is gently forced down onto a specimen. As it moves across the surface of the specimen, its movements are recorded to produce a 3D image.   Preparing Smears for staining Staining – Coloring specimen with dyes that emphasize certain structures. Smear – Thin films of material containing microorganisms spread over the slide.  Fixed – (Attached) Passing specimens through the flame of a Bunsen burner serveral times.  Basic dyes – stain composed of a Cation in chromophore Acidic dyes – stain composed of the Anion in chromophore.  Negative staining – Preparing colorless bacteria against a colored background.  Simple stain – An aqueous or alcohol solution of a single basic dye.  Differential stains – React differently with different kinds of bacteria and thus can be used to  distinguish them.  Gram stain –Classifies bacteria into two large groups, gram positive and gram negative.  Bacteria that retains the dark violet color after decolorization are gram­positive, bacteria that lose the color are gram­negative.  Acid fast stains – Binds strongly only to bacteria that have a waxy material in their cell walls.  ­Used to identify mycobacterium/nocardia Special stains – Used to color parts of microorganism such as flagella, endospheres, or capsules. 


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