Chapter 3: Observing microorganism through a microscope Flashcards
ocular lens
- remagnifies the image fromed by the objective lens
body tube
- transmits the image from the objective lens to the ocular lens
Objective lens
- primary lens that magnifies the speciman
Stage
- holds the microscope slide in position
condenser
- focuses light through specimen
Diapragm
- controls the amount of light entering the condenser
Illuminator
- light source
what would be the total magnification of a compund microscope with the objective lens magnification of 40x and the ocular lens of 10x
- 400x
3 types of stain
- simple
- differnetial
- special
simple stain
- aqueous or alchol solution made fo a single basic dye
-uses a moderate
purpose of simple stain
- distinguish cellular shapes or structure
Mordant
- may be used to hold stain or coat the specimen to enlarge it
Differitial stain purpose
- distinuish the type of bacteria
types of differntial stains
- gram
- acid fast
Gram Stain positive process
- primary stain (cystal violet): purple
- Moderant (Iodine): purple
- Decolorizing Agent (alchol): purple
- Counterstain (safferin): purple
Gram stain negative process
- primary stain (cystal violet): purple
- Moderant (Iodine): purple
- Decolorizing Agent (alchol): colorless
- Counterstain (safferin): red
why is there a difference in the colors of gram stain
- because of the diffrent makeup of the cell wall
Acid fast stain purpose
- holds strong to bacteria that have waxy material
- used to identify mycobacterium species
Acid fast positive process
- primary stain (carbonfusion): red
- Decolorizing agent (acid-alchol): red
- counterstain (methylene blue): red
Acid fast stain negative process
- primary stain (carbonfusion): red
- Decolorizing agent (acid-alchol): colorless
- counterstain (methylene blue): blue
3 types of special stains
- glycocalyx
- endospores
- flagella
Glycocalyx process
- acidic stain: india ink or nigrosin
- water wash (PROBLEM: water soluable capsule so have to be gental)
- saffranin counterstain
Endospores stain process
- malachite green
- water wash
- counterstain: saffrinin
flagella staining process
- carbolfuchsin (red)
- Mordant —> potassium alum (dark red)
what is a stain?
- coloring the microorganism with dye that emphasizes certain structures
Chromophore stain
- stains are salts; compromised of a positive and negative ion –> one of which is coloured
Acidic dyes
- is the anion
Basic dyes
- is the cation
- used for bacteria
- used most frequently
why are basic dyes used for bacteria
- bacteria is slightly negativly charged therefore the cation basic dye is attracted to it
Examples of basic dye
- crystal violet
- methylene blue
- malachite green
- safffrin
negative staining
- staining the background instead of the cel
- useful for observing overall cell shapes, capsules, and sizes becasue distortion are limited