Instructions for Making On-Line Reservations for Microscopes in NB11-204 1. Log into Mail using Mail.swmed.edu 2. Log in using your university id and password. 3. Click the Calendar Tab at the top right of the screen. 4. Click MolePathScope Sign Up at the left of the screen. 5. Click New Event at the top left-hand corner of the screen. 6. In the Event line on the new screen enter the scope you wish to use (Compound, Inverted or Dissection). Also indicate if you will be using bright-field illumination or fluorescence. 7. On the Attendees Line write MolePathScopeSignUp. 8. Enter date and start time. 9. The maximum time allowed is 2 hours. When finished click Send at the upper left-hand corner of the screen.
Operating Instructions Leica Microscope Room NB11-204 Bright field 1. Turn the power on using orange switch on the right-hand side of the base. 2. Pull the Beam Splitter on the upper left-hand side of the scope to 50/50 or Vis. 3. Adjust the light level to 8 by adjusting the Bright-Field Rheostat on the lower left-hand side of the base. 4. Use the correct condenser. One condenser will be mounted on the scope, the other is stored in the top drawer immediately to your left. a. For low magnification objectives (1.25X, 2.5X, 4X or 10X) use the Nikon Achro 0.13 condenser b. For high magnification (20X, 40X, 63X or 100X) use the Leica 0.9 PSI. The condenser is stored in the top drawer to your left. c. To replace the condenser, lower the condenser to the lowest Z positon using the Condenser Z-Axis Focus knob behind the condenser and loosen the silver grub screw on the right side of the condenser and remove the condenser towards you. 5. Replace the chosen condenser by sliding it into the support bracket. When using the high-magnification condenser, be careful to insert the pin on the back of the condenser into the notch at the back of the support. 6. Tighten the grub screw to secure the condenser in place. 7. Establish Kohler Illumination for the high-magnification condenser. a. Raise the condenser to near the top limit without hitting the slide. b. Place a slide on the stage to establish plane of focus using the 20X or 40X objective.
c. Close the Field Diaphragm so that you can see it through the objective. d. Adjust the Z axis of the condenser to bring the edge of the leaves of the field diaphragm into focus. e. Open the field diaphragm just beyond the limits of the field of view and then turn the Condenser Iris to match the color of the ring on objective you are using. 8. When using the low-magnification condenser, position it at the lowest Z position and open the Condenser Iris Diaphragm completely. To Acquire Data Log into computer by clicking on Camera User. Use Password: Password123. To save you files, open Documents, then Work, then open your lab folder.
Operating Instructions Leica Microscope Room NB11-204 Fluorescence 1. Turn the bright-field light source off using orange Power Switch situated at the base on the right side. 2. Feel the X-Cite lamp housing. If it is hot wait ½ hour before using the lamp. 3. Turn the power on using the Illuminator Power Switch. 4. Check the following: a. Illuminator Iris Diaphragm on the lamp housing is open. b. All four light path levers on the right side of the scope housing are adjusted to the top. c. Fluorescence Filter Wheel at the front of the scope is set to position 2. 5. If all is well you now will see blue light coming through the lens. 6. Use 20X or higher objectives. 7. If the emission is dim then pull or push the Beam Splitter (upper left side of the scope housing) to Vis, or if acquiring an image set to Photo. 8. Adjust the Fluorescence Filter Wheel to excite the specific fluorophore you are using. a. #1 UV for Dapi, Hoechst, AMCA b. #2 GFP, FITZ, Alexa 488, Cy2 c. #3 Cy3, Texas Red, Rhodamine, Alexa 525 9. To improve emission, keep the condenser lowered or flipped forward using the lever on the left of the condenser housing. 10. Leave the X-Cite illuminator on for a minimum of 30 minutes. To Acquire Data Log into computer by clicking on Camera User. Use Password: Password123. To save you files, open Documents, then Work, then open your lab folder.