1. MIKROSCOPE 2. Haemositometer: Manual, software Dr. Gatot Ciptadi Lab.Genetika-Pemuliaan ternak...
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Transcript of 1. MIKROSCOPE 2. Haemositometer: Manual, software Dr. Gatot Ciptadi Lab.Genetika-Pemuliaan ternak...
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MIKROSCOPE
2. Haemositometer:Manual, software
Dr. Gatot CiptadiLab.Genetika-Pemuliaan ternak
Lab Sentral Ilmu Hayati (LSIH)-UB
Introduction to the Microscope:CarePartsFocusing
1m = 103 mm (millimetres)1m = 106 µm (micrometres)
1m = 109 nm (nanometres)
UNITS OF MEASUREMENT
Eyepiece
Body Tube
Revolving Nosepiece
ArmObjective Lens
StageStage Clips
Coarse Focus
Fine Focus
Base
Diaphragm
Light
• Always carry with 2 hands• Only use lens paper for
cleaning• Do not force knobs• Always store covered• Keep objects clear of desk
and cords
•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.
•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.
Compound Light MicroscopeCompound Light Microscope
• 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 mikroskopMacam-macam mikroskop(berdasarkan pencahayaannya)(berdasarkan pencahayaannya)
1.1. Mikroskop cahayaMikroskop cahaya
2.2. Mikroskop stereoMikroskop stereo
3.3. Mikroskop elektronMikroskop elektron
Dissecting Microscope
Dissecting MicroscopeDissecting 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.
•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.
4 – When you are done, turn off the microscope and put up the slides you used.
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!
Carrying a Microscope
1. Rotate the low power objective into place and make sure the stage is all the way down.
2. 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. Turn light on.
4. Focus first with the coarse adjustment knob. Once in focus on low power, turn the nosepiece until the next
higher lens is in place.
5. Use FINE adjustment knob ONLY and focus the object.
Steps to Use:
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.
You do not need to use the stage clips when viewing wet-mount slides!
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.
4 - Slowly lower the cover slip on top of the drop.
Cover Slip
Lower slowly
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 elektronMikroskope elektron• Mikroskop elektron mempunyai Mikroskop elektron mempunyai 33 tipe, tipe,
•
– mikroskop elektron scanning mikroskop elektron scanning (SEM) (SEM) untuk studi detil untuk studi detil arsitektur permukaan sel (atau arsitektur permukaan sel (atau struktur renik lainnya), dan obyek struktur renik lainnya), dan obyek diamati secara tiga dimensi. diamati secara tiga dimensi.
– mikroskop elektron transmisi mikroskop elektron transmisi (TEM)(TEM) mengamati struktur detil mengamati struktur detil internal sel. internal sel.
– Scanning Tunneling Microscope ( (STM) ) untuk melihat permukaan untuk melihat permukaan atomatom
Scanning Electron Microscope (SEM)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 ???
•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)
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.
•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
fluorescent (TV) screen,photographic film
Human eye (retina), photographic film
Focussing screen
VacuumAir-filledInterior
MagnetsGlassLenses
High voltage (50kV) tungsten lamp
Tungsten or quartz halogen lamp
Radiation source
x500 000x1000 – x1500Maximum
magnification
0.2nmFine detail
app. 200nmMaximum resolving power
Electronsapp. 4nm
Monochrome
Visible light760nm (red) – 390nm
Colours visible
Electromagnetic spectrum used
ELECTRON MICROSCOPE
LIGHT MICROSCOPEFEATURE
© 2007 Paul Billiet ODWS
THE LIGHT MICROSCOPE v THE ELECTRON MICROSCOPE
Copper gridGlass slideSupport
Heavy metalsWater soluble dyesStains
Microtome only.Slices 50nm
Parts of cells visible
Hand or microtomeslices 20 000nmWhole cells visible
Sectioning
ResinWaxEmbedding
OsO4 or KMnO4AlcoholFixation
Tissues must be dehydrated
= dead
Temporary mounts living or dead
Preparation of specimens
ELECTRON MICROSCOPE
LIGHT MICROSCOPEFEATURE
© 2007 Paul Billiet ODWS