Transcript fine adjustment knob

MIKROSCOPE
1. Introduction to the Microscope:
 Care
 Parts
 Focusing
2. Haemositometer:
Manual, software
Dr. Gatot Ciptadi
Lab.Genetika-Pemuliaan ternak
Lab Sentral Ilmu Hayati (LSIH)-UB
UNITS OF MEASUREMENT
1m = 103 mm (millimetres)
1m = 106 µm (micrometres)
1m = 109 nm (nanometres)
Body Tube
Eyepiece
Revolving Nosepiece
Objective Lens
Stage
Clips
Diaphragm
Light
Arm
• Always carry with 2 hands
• Only use lens paper for
Stage
cleaning
Coarse Focus
• Do not force knobs
Fine Focus
• Always store covered
• Keep objects clear of desk
Base
and cords
Compound Light Microscope
•Light shines through specimen and into
a single objective lens and then through
the eyepiece (ocular).
•Provides two-dimensional view.
•Specimen must be thin and light must be
able to pass through.
•Useful up to about 1,000 X
magnification.
•
•
•
•
•
Place the Slide on the Microscope
Use Stage Clips
Click Nosepiece to the lowest (shortest) setting
Look into the Eyepiece
Use the Coarse Focus
• Follow steps to focus using low
power
• Click the nosepiece to the longest
objective
• Do NOT use the Coarse Focusing
Knob
• Use the Fine Focus Knob to bring the
slide
Macam-macam mikroskop
(berdasarkan pencahayaannya)
1.
2.
3.
Mikroskop cahaya
Mikroskop stereo
Mikroskop elektron
Dissecting Microscope
Dissecting Microscope
•Light is reflected off a specimen into two
different objectives and eyepieces (or
oculars).
• Provides a three-dimensional view of
object.
•Usually shows surface features of object.
•Useful up to about 60 - 80 X
magnification.
CLSM
Cultured Cell
Live Cell Imaging
Let’s give it a try ...
1 – Turn on the microscope and then rotate the nosepiece to click the
red-banded objective into place.
2 – Place a slide on the stage and secure it using the stage clips. Use
the coarse adjustment knob (large knob) to get it the image into view
and then use the fine adjustment knob (small knob) to make it
clearer.
3 – Once you have the image in view, rotate the nosepiece to view it
under different powers. Draw what you see on your worksheet!
Be careful with the largest objective! Sometimes there is
not enough room and you will not be able to use it!
4 – When you are done, turn off the microscope and put up the
slides you used.
Carrying a Microscope
Steps to Use:
1.
2.
Rotate the low power objective into place and make
sure the stage is all the way down.
Place slide on stage making sure object to be viewed is
centered over the hole in the stage. Use the stage clips
to hold the slide in place.
3.
4.
5.
Turn light on.
Focus first with the coarse adjustment knob. Once in
focus on low power, turn the nosepiece until the next
higher lens is in place.
Use FINE adjustment knob ONLY and focus the
object.
How to make a wet-mount slide …
1 – Get a clean slide and coverslip.
2 – Place ONE drop of water in the middle of the slide. Don’t use
too much or the water will run off the edge and make a mess!
3 – Place the edge of the cover slip on one side of the water drop.
4 - Slowly lower the cover slip on top of the drop.
Cover
Slip
Lower slowly
5 – Place the slide on the stage and view it first with the red-banded
objective. Once you see the image, you can rotate the nosepiece to
view the slide with the different objectives.
You do not need to use the stage clips
when viewing wet-mount slides!
INVERTED MICROSCOPE
• Inverted microscope is
widely used for direct
observation of cells in
cultivation flasks.
• Observe various cell cultures
using this microscope.
Compare their morphology
and density.
Mikroskope elektron
• Mikroskop elektron mempunyai 3 tipe,
•
– mikroskop elektron scanning
(SEM)  untuk studi detil
arsitektur permukaan sel (atau
struktur renik lainnya), dan obyek
diamati secara tiga dimensi.
– mikroskop elektron transmisi
(TEM)  mengamati struktur detil
internal sel.
– Scanning Tunneling Microscope
(STM)  untuk melihat permukaan
atom
Scanning Electron Microscope (SEM)
•Uses a beam of electrons instead of
light.
•The beam of electrons is passed
over the specimen and are scattered.
These scattered electrons are
detected and processed to form an
image on a florescent screen.
•Useful up to about ???
Scanning Tunneling Electron
Microscope (TEM)
•Uses a beam of electrons
instead of light.
•Uses probe and electrons
to determine differences
in voltage as probe passes
over specimen.
•Can view objects as
small as atoms.
UB ?
1. Inverted Mikroskop dan mikromanipulator:
FMIPA-BIO, Lab sentral, dan Kedokteran
2. Confocal Laser Scanning Microscope (CLSM):
Lab sentral UB; 1. 3 Milyard.)
THE LIGHT MICROSCOPE v THE ELECTRON MICROSCOPE
FEATURE
LIGHT MICROSCOPE
ELECTRON
MICROSCOPE
Electromagnetic
spectrum used
Visible light
760nm (red) – 390nm
Colours visible
Electrons
app. 4nm
Monochrome
Maximum
resolving power
app. 200nm
0.2nm
Fine detail
Maximum
magnification
x1000 – x1500
x500 000
Radiation
source
Tungsten or quartz
halogen lamp
High voltage (50kV)
tungsten lamp
Lenses
Interior
Glass
Air-filled
Magnets
Vacuum
Focussing
screen
Human eye (retina),
photographic film
fluorescent (TV) screen,
photographic film
© 2007 Paul Billiet ODWS
THE LIGHT MICROSCOPE v THE ELECTRON MICROSCOPE
ELECTRON
MICROSCOPE
Tissues must be
dehydrated
= dead
FEATURE
LIGHT MICROSCOPE
Preparation of
specimens
Temporary mounts
living or dead
Fixation
Embedding
Alcohol
OsO4 or KMnO4
Resin
Sectioning
Wax
Hand or microtome
slices  20 000nm
Whole cells visible
Microtome only.
Slices  50nm
Parts of cells visible
Stains
Water soluble dyes
Heavy metals
Support
Glass slide
Copper grid
© 2007 Paul Billiet ODWS