2. 1. MIKROSCOPE 2. Haemositometer: Manual, software Dr. Gatot Ciptadi Lab.Genetika-Pemuliaan ternak Lab Sentral Ilmu Hayati (LSIH)-UB Introduction to the Microscope:  Care  Parts  Focusing

3. 1m = 10 3 mm (millimetres) 1m = 10 6 µm (micrometres) 1m = 10 9 nm (nanometres ) UNITS OF MEASUREMENT

4. Eyepiece Body Tube Revolving Nosepiece Arm Objective Lens Stage Stage 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

5. 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 Microscope

6. 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

7. Macam-macam mikroskop (berdasarkan pencahayaannya) 1.Mikroskop cahaya 2.Mikroskop stereo 3.Mikroskop elektron Dissecting Microscope

8. 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.

9. CLSM Cultured Cell Live Cell Imaging

10. 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!

11. 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:

12. 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

13. 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.

14. Mikroskope elektron Mikroskop elektron mempunyai 3 tipe,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 Tunneling MicroscopeSTMScanning Tunneling MicroscopeSTM

15. 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 ???

16. 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.

17. UB ? 1.Inverted Mikroskop dan mikromanipulator: FMIPA-BIO, Lab sentral, dan Kedokteran 2. Confocal Laser Scanning Microscope (CLSM): Lab sentral UB; 1. 3 Milyard.)

18. 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 – x1500 Maximum magnification 0.2nm Fine detail app. 200nmMaximum resolving power Electrons app. 4nm Monochrome Visible light 760nm (red) – 390nm Colours visible Electromagnetic spectrum used ELECTRON MICROSCOPE LIGHT MICROSCOPEFEATURE © 2007 Paul Billiet ODWSODWS

19. THE LIGHT MICROSCOPE v THE ELECTRON MICROSCOPE Copper gridGlass slideSupport Heavy metalsWater soluble dyesStains Microtome only. Slices  50nm Parts of cells visible Hand or microtome slices  20 000nm Whole cells visible Sectioning ResinWax Embedding OsO 4 or KMnO 4 AlcoholFixation Tissues must be dehydrated = dead Temporary mounts living or dead Preparation of specimens ELECTRON MICROSCOPE LIGHT MICROSCOPEFEATURE © 2007 Paul Billiet ODWSODWS