Microscopy Flashcards
Intro:
- _____ published hai treatise _________
- included: ______ & ______ - microscope contains single bi-convex magnifying lens
- thicker in the ____ than _____
- limited resolution & magnifying power due to _____
- most commercial magnifiers are able to produce ×____ - with two lens system
- founder
- 1st lens system
- 2nd lens system
- obj lens projects ____
- eyepiece magnifies ______
- Hooke; Micrographica
- illustrations of mold forms & anatomy of the flea - Simple Microscope
- center than peripheral
- low numerical aperture
- 2-30x magnification - Compound Microscope
- Zaccharias Janssen & Hans Janssen
- objective lens
- ocular lens
- magnified image
- projected image
TYPES OF MICROSCOPE
- creating a magnified image thru the use of a series of glass lenses
- uses ___ to detect smaller object - most advanced & important type of microscope with highest magnifying capacity
- uses ___ to illuminate tiniest particles
- Light Microscope
- visible light - Electron Microscope
- electrons
TYPES OF LIGHT MICROSCOPE
_____: most commonly used light microscope
- forms a ___ image against ___ background
- ______ illumination
Parts of the Bright-Field Microscope:
1. focuses the light into the specimen and evenly
distribute the light from the source
2. regulates the diameter of light beams that will
strike the specimen
- adjusts the ______
3. mechanical stage knob:
- top
- bottom
4. regulates light intensity
Brightfield Microscope
- dark image against bright background
- Kohler
Parts of the Bright-Field Microscope:
1. Condenser
2. Iris diaphragm
- aperture size
3. mechanical stage knob:
- forward/backward
- left/right
4. Rheostat
TYPES OF LIGHT MICROSCOPE
_____: condenser does not allow light to pass directly thru specimen
- added to block the light that forms an oblique angle
- used to view spirochetes (_____)
Darkfield Microscope
- dark-field ring
- Treponema pallidum
TYPES OF LIGHT MICROSCOPE
_____: allows visualization of naturally fluorescent microorganisms or those microscopes with highest magnifying capacity; more powerful stained by fluorescent dye (antibody)/ fluorophores fluorochromes
- exposes the specimen to ____
- involves the excitation of fluorochromes using lights
- selects specific wavelength/illumination which will be absorbed by the specimen
- transmits different wavelength that is emitted from specimen to eyepiece - dyes: (3)
- for fungi
- FITC - direct interaction
- with antigen
- hard to cultivate bacteria’s (3)
Fluorescent Microscope
- UV light
2.
- excitation filter
- barrier filter - acridine orange, auramine, rhodamine
- calcofluor white
- Fluoroscein isothiocyanate
4.Fluorochroming
5.Immunofluorescence
6.
- Legionella
- Bordeteklla
- Chylamadia
TYPES OF LIGHT MICROSCOPE
_____: light beams are deflected by diff. thickness of the object; light beams
are reflected a second time when they strike the objective
- based on
- used to examine ____ cells
- for ___ prep
- ____ is added - permits detailed examination of internal structures in living
organism = detection of bacterial components (____ & ______)
- contains ____, ____, & _____ - opaque disk with a thin transparent
ring which produces a hollow cone of light - background is ___ while the unstained object appears ____
- ___ can be converted phase contrast by using ______
Phase Contrast Microscope
- refractive index
- unstained living cells
- wet mount prep
- saline - endospore & inclusion bodies
- β–hydroxy butyrate,
polymetaphosphate, sulfur - annular stop
- bright; dark
- brightfield microscope ; anula rings
Types of Electron Microscipe
- electrons pass directly thru the specimen (hence, can visualize
internal structures of cell)
- magnification
- more _____ form of EM
- ___ of microorganisms
- stain ___ or ____ - electrons strikes surface of specimen
- magnification
- more _____ form of EM
- ___ of microorganisms
- stain ___ or ____ - shows three-dimensional images of a sample
- studied using ____ that scans the surface
- electrons tunnel between surface & stylus produces ____
- ___ movement of stylus is recorded
- Transmission Electron Microscope (TEM)
- >100,000x
- traditional
- ultrathin slices
- gold or palladium - Scanning Electron Microscope (SEM)
- >1,000x
- contemporary
- Whole objects
- gold or palladium - Scanning Tunneling Microscope (STM)
- stylus
- electrical signal
- vertical
TERMINOLOGIES
- process of enlarging an object’s size without
affecting its actual size
- total magnification formula - specific point at which parallel rays of light
converge after passing through a lens - distance from the lens center to a point where
parallel rays are focused on the optical axis
- shorter FL
- longer FL - important factor
- shorter wavelength = ____ the resolution
- use ___ of visible spectrum (____nm) - distance between 2 front surface of the lens and the
surface of cover glass (if one is used) or specimen when it is in sharp focus
- objectives w/ large numerical apertures and great resolving powers have _____
- Magnification
- ocular x objective
magnification - Focal Point
- Focal Length
- stronger lens
- weaker lens - Wavelength
- higher
- blue end (450-500nm) - Working Distance
- short working
TERMINOLOGIES
_____: ability of a lens to distinguish two points or objects as separate
1. aka
2. he formulated this equation
3. equation which shows the maximum resolution for microscope
4. formula
- wavelength of light
- numerical aperture
- min distance between 2 objects
5. as d becomes ___ = resolution _____
RESOLUTION
- RESOLVING POWER
- Ernst Abbe
- Abbe sine equation
- formula
- lambda
- n sin ø
- d - as d becomes smaller = resolution increases
TERMINOLOGI
_____: number that indicates resolving power of lens system
- ½ the angle of the cone of light entering an
objective - refractive index
- n of air is ___
- no lens working in air can have NA of ___
- to increase the refractive index, use ____
- maximum theoretical resolving power of a
microscope with an:
- OIO
- blue green lights - typical NA values
- 4x
- 10x
- 40x
- 100x
NUMERICAL APERTURE
- Ø
- N
- 1.00
- > 1
- IMMERSION OIL
6.
- 1.25
- 0.2 - typical NA values
- 0.1
- 0.25
- 0.65
- 1.25
