Page 3 - Turning on the microscope
1 1 General user information Leica DM LB research microscope is only for the Molecular Cancer Biology (MCBP) and Genome-scale Biology (GSB) research programs internal use. It is a transmitted light microscope that cannot be used for fluorescence microscopy. Bright-field, polarization contrast, and l...
Page 5 - Focusing
3 Objective Numerical Aperture (NA) Color ring Coverglass (c.g.) thickness Position for auxiliary condensor lens N Plan 2.5x 0.07 Brown Any/without c.g. off N Plan 5x 0.12 Red Any/without c.g. off N Plan 10x 0.25 Yellow Any/without c.g. on N Plan 20x 0.4 Green 0.17 mm on N Plan 40x 0.65 Blue 0.17 mm...
Page 6 - Dual viewing attachment
4 The fine focus has two modes: when the left focus wheel is pulled out, each line on the wheel equals 1 µ m of movement. When it is pushed in, each line equals 4 µ m of movement. Figure 5. Focusing wheels 2.6 Eye pieces (oculars) Adjust the folding bridge of the oculars until you see, using both ey...
Page 7 - Move up or down to control position horizontally
5 Figure 7. Dual viewing attachment Figure 8. Power source for the light pointer There is a light pointer in the viewing attachment that uses an external power source. Thus, you need to connect the transformer into a power outlet before using the pointer (Figure 8). After use, disconnect the transfo...
Page 8 - Koehler illumination; Instruction
6 Figure 9. Light pointer controls 2.8 Koehler illumination Koehler illumination needs to be adjusted in the beginning of every imaging session for optimal illumination. Proceed as follows: Instruction View in eyepiece 1. Use 10x objective and front lens. Focus on the sample. 2. Close the luminous-f...
Page 9 - condenser
7 Instruction View in eyepiece 4. Center the diaphragm in the field of view by rotating the condenser adjusting screws. In other words, you adjust the illuminating path so that it is properly aligned along the optical axis of the microscope. 5. Open the luminous-field diaphragm by turning the adjust...
Page 10 - Polarization contrast; Filters
8 2.9 Polarization contrast Figure 10. Analyzer for polarization contrast imaging Figure 11. Polarizer For polarization contrast microscopy, an analyzer (a polarizing filter) is inserted in the light path by pushing it all the way in (Figure 10), and another polarizing filter (polarizer) is placed l...
Page 11 - Dark-field microscopy; Turning off the microscope; When you have finished using the microscope you should:
9 • Grun: a green panchromatic filter that was used for contrast enchantment for B/W photography These filters were used for film photography and are not useful for digital photography. However, when using low intensity light (yellowish), DLF-filter adjusts the light back to the direction of blue, m...
Page 12 - Image acquisition; Accessing the computer; Adjustments before image acquisition; Capture resolution
10 3 Image acquisition 3.1 Accessing the computer If the computer is turned off, push the power button to turn it on. When the login screen appears, log in using your hyad login account. Check that the domain is set to LTDK. 3.2 View Finder application A 10-bit Olympus DP50 color CCD camera is attac...
Page 14 - Focus tool; Averaging
12 Figure 16. Histogram level adjustment. Red square: levels button 3.3.6 Focus tool Focusing tool assists you in focusing. If the focusing toolbar is not visible, enable it in the Options menu. Once activated, two bars appear (Figure 17). The blue one shows the current focus value, and the red one ...
Page 15 - Post-imaging adjustments; Cropping the image
13 15). Make sure the stage does not move at all between the images or image quality decreases. 3.4 Capturing the image Press the Camera button next to the Live button to take an image. After acquisition, the image will appear in Studio Lite. 3.5 Viewing images To zoom in or out, select the zoom too...
Page 16 - Scale bars; Saving and transferring images
14 3.6.2 Scale bars The Studio Lite software does not provide means to insert scale bars. They can be added later on in any image editing software such as Adobe Photoshop. Ready-made scale bars for the full resolution images are available at http://www.miu.helsinki.fi/instruments/LeicaDMLB/scale_pic...
Page 17 - Troubleshooting
15 • Cover the microscope with the red hood 6 Troubleshooting 6.1 There is no light • Microscope is not turned on (see Section 2.1) • Light intensity set too low (see 2.1 on page 1) • Do you see any light in the lamp chamber? If not, the halogen bulb may need to be changed - contact MIU 6.2 Light is...
Page 18 - Computer login fails; domain is LTDK
16 6.7 Computer login fails Check that: • domain is LTDK • use are using your hyad account user ID and password - the same ones you are using for accessing your university e-mail account