ANAPHY LAB MICROSCOPY L1 Flashcards
7 DEFINITION OF TERMS OF MICROSCOPY
- MAGNIFICATION
- RESOLUTION/RESOLVING POWER
3.NUMERICAL APERTURE
4.FOCAL LENGTH
5.WORKING DISTANCE
6.PARFOCAL
7.REFRACTIVE INDEX
Bending of light rays away from the objective lens when light passes from the glass of the
microscope slide to the air.
Refractive Index
Refers to quality of the objectives & eyepiece where practically no change in focus has to be
made when objective is substituted for another
PARFOCAL
Distance between the front lens of the objective lens & the top of the cover glass when the
specimen is in focus
WORKING DISTANCE
- Thickness of the object that maybe seen at one time under focus.
- Distance between the center of a lens or curved mirror and its focus
FOCAL LENGTH
A measurement of the ability of the condenser and the objective lens to gather light.
NUMERICAL APERTURE
MEANING OF LPO HPO OIO
LOW POWER OBJECTIVE
HIGH POWER OBJECTIVE
OIL IMMERSION OBJECTIVE
SCANNER
MAGNIFICATION OF SCANNER AND NUMERICAL APERTURE
4X; 0.10
MAGNIFICATION OF LPO AND N.A
10X; 0.25
MAGNIFICATION AND N.A OF HPO
40X; 0.65
MAGNIFICATION AND N.A OF OIO
100X; 1.25
4 TYPES OF MICROSCOPE
SIMPLE
COMPOUND
SCANNING E
TRANSMISSION E
Created in the 17th century by Antony van
Leeuwenhoek, who combined a convex lens with a
holder for specimens.
200 – 300x Magnification
SIMPLE
Two lenses
Better magnification than a simple microscope
COMPOUND MICROSCOPE
the specimen is lit from
underneath, and they can be binocular or monocular)
provide a magnification of 1,000 times, which is
considered to be high, although the resolution is low.
BRIGHT FIELD
Stained tissue is examined with ordinary light
passing through the preparation.
BRIGHT FIELD
TYPES OF COMPOUND MICROSCOPE
BRIGHT FIELD MICROSCOPE
FLUORESCENCE MICROSCOPY
PHASE-CONTRAST MICROSCOPY
POLARIZING MICROSCOPY
Tissue sections are usually irradiated with
ultraviolet (UV) light and the emission is in the
visible portion of the spectrum.
FLUORESCENCE MICROSCOPY
Fluorescent substances appear bright on a dark
background
FLUORESCENCE MICROSCOPY
uses a lens system that produces visible images
from transparent objects and, importantly, can be
used with living, cultured cells
PHASE-CONTAST MICROSCOPY
Allows the recognition of stained or
unstained structures made of highly
organized subunits.
POLARIZING MICROSCOPY
Tissue structures containing oriented
macromolecules are located between the
two polarizing filters, they appear as
bright structures against a dark
background.
POLARIZING MICROSCOPY
type of microscope that uses visible light and an objective lens to magnify and illuminate a sample.
BF MICROSCOPY
Type of optical microscopy where a fluorescent sample is illuminated with light of certain wavelength
FLUORESCENCE
a light microscopy technique used to enhance the contrast of images of transparent and colourless specimens.
BLACK AND WHITE
PHASE-CONTRAST
a microscope that utilizes polarized light to reveal detail in an object, used especially to study crystalline and fibrous structures.
POLARIZING
- Uses electrons rather than light for image
formation. - Samples are scanned in vacuum or near-
vacuum conditions, so they must be specially
prepared by first undergoing dehydration and
then being coated with a thin layer of a
conducive material, such as gold. - After the item is prepared and placed in the
chamber, the SEM produces a 3-D, black-and-
white image on a computer
SEM
-Uses electrons in creating a magnified image
- Uses a slide preparation to obtain a 2-D view
of specimens, so it’s more suited for viewing
objects with some degree of transparency. - Offers a high degree of both magnification and
resolution
TEM
3 GEN PART OF MICROSCOPE
MECHANICAL
MAGNIFYING
ILLUMINATING
used to support and adjust the parts
MECHANICAL
USED TO ENLARGE SPECIMEN
MAGNIFYING
USED TO PROVIDE LIGHTS
ILLUMINATING
post portion that supports the entire
microscope, provides stability and support for the
microscope when it is upright. also typically
holds the illuminator, or light source
BASE
part above the base that supports the other parts
PILLAR
allows for tilting for the convenience
of the user
INCLINATION JOINT
connects the eyepiece tube to the base ,curve or
slanted part which is held while carrying the
microscope
ARM - CARRYING HANDLE
platform for the slides,
which hold the specimen. The stage typically has a
stage clip on either side to hold the slide firmly in
place. Some microscopes have a mechanical stage,
with adjustment knobs that allow for more precise
positioning of slides
Stage and stage clip
attached to the arm and bears the lenses
BODY TUBE
cylindrical structure on the top of the
body tube that holds the ocular lenses
Draw tube
contains the objective
lenses. Microscope users can rotate this part to
switch between the objective lenses and adjust the
magnification power
REVOLVING NOSE PIECE
Lies atop the nose piece and keeps dust
from settling on the objectives
DUST SHIELD
moves
stage (or body tube) up and down, used
to focus the image on the
microscope under low or medium
power
Coarse Adjustment Knob
small knob,
used to fine-tune the focus of your
specimen after using the coarse
adjustment knob
Fine Adjustment Knob
ontains the ocular lens,
which the user looks through to see the
magnified specimen, further magnify the
image produced by the objective lenses by 5x
to 15x
EYEPIECE/OCULAR
combine with the eyepiece
lens to increase magnification levels.
Microscopes generally feature three or four
objective lenses, with magnification levels
ranging 4x to 100x
OBJECTIVE LENSES
light source for viewing. This can come
in the form of a built-in, low-voltage illuminator
light, or a mirror that reflects an external light
source like sunlighT
ILLUMINATOR/ E-LAMP
has a concave and plane surface to
gather and direct light in order to illuminate the
object
MIRROR
focus the light onto the specimen,
concentrating light rays on the specimen
CONDENSER
controls the amount of light
reaching the specimen and regulates the light
necessary to obtain a clear view of the objecT
IRIS DIAPHRAGM
USING MICROSCOPE
The proper way to focus a microscope is to start with the lowest power objective lens first
and while looking from the side, crank the lens down as close to the specimen as possible
without touching it.
* Look through the eyepiece lens and focus upward only until the image is sharp. If you can’t
get it in focus, repeat the process again.
* Once the image is sharp with the low power lens, you should be able to simply click in the
next power lens and do minor adjustments with the focus knob.
* If your microscope has a fine focus adjustment, turning it a bit should be all that’s
necessary. Continue with subsequent objective lenses and fine focus each time.
CARE OF THE MICROSCOPE
-Always carry with 2 hands
* Never touch the lenses with your fingers.
* Only use lens paper for cleaning
* Keep objects clear of desk and cords
* When you are finished with your “scope”, rotate the nosepiece so that it’s
on the low power objective, roll the stage down to lowest level, rubber
band the cord, then replace the dust cover.
