Lab Quiz #1

Beschreibung

This includes exercises 2, 6, 7, and 8 which are entitled The Compound Light Microscope; The Negative Stain; Preparing a Bacterial Smear; and The Simple Stain
Calla Frederiks
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Zusammenfassung der Ressource

Frage Antworten
Microscope an invaluable tool used for observing objects that can not be seen by the naked eye; enables us to view the ultrastructure (fine detail) of organisms, cells, and tissues.
compound light microscope magnifies objects from 40 to 1000 times its original size specimens must be either small objects of very thinly cut sections of an object; translucent allowing light to be transmitted through them
dissecting (stereoscopic) microscope magnifies objects from 4 to 50 times specimens are usually opaque
Normal Procedure to prepare a specimen for most staining protocols allow the organisms to dry to a glass slide; pass the slide through the Bunsen burner flame in order to fix the organisms to the slide
Problems with this procedure and why This procedure does not allow one to visualize the cells in their natural state. The size and shape of the cell may become distorted. Also, heat-fixing removes the water from the cell which affects the size of the cell when viewed under the microscope
Pros to Negative Staining allows one to view living organisms that have not been heat-fixed to a slide; allows one to visualize organisms or parts of organisms, such as a capsule, which may not readily take up stains
Negative Staining characteristics The background is filled with a stain (such as India ink or nigrosin) while the organisms remain clear, unstained objects that stand out clearly against this dark background.
The "why" behind the cells appearing clear when stained These stains (negative stains) do not penetrate the bacterial cells due to the repulsion between the negative charge of the stains and the negatively charged bacterial cell.
Two different methods to prepare negative stains a suspension of bacterial cells is emulsified in a drop of nigrosin (India ink) and covered with a cover slip (wet mount) or allowed to air dry (smear)
Most microorganisms appear ______ when viewed through a ___________ _______ microscope. colorless; brightfield light
Most common method method of allowing visualization of microorganisms under a microscope is to ... stain, or color them.
Staining _________ to the microorganisms so that various structures can be observed imparts color
How to prepare a bacterial smear A drop of liquid bacterial culture or a drop of water and a portion of a bacterial colony is spread over the surface of the slide
Important facts about preparing the bacterial smear one loopful of liquid bacterial culture usually provides sufficient bacteria for easy viewing REMEMBER 1 bacterial colony = millions of individual bacteria When mixing water with the bacteria only a small drop of water is needed
Why is only a small amount of water needed? the smears are air-dried, the more liquid, the longer the smear will take to dry
When mixing the bacteria with the water to create the smear... avoid overmixing because this can lead to inaccurate bacterial cell patterns (chains may appear as clumps and vice versa)
Once the smear has air-dried, the next step is to heat-fix the slide
Heat-fix The air-dried smear is briefly passed through the Bunsen burner flame 2 or 3 times.
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