Zeiss ICM 405 Inverted Fluorescence Microscope Operating Instructions

Remove cover

2 Turn on Mercury Vapour Lamp, toggle switch on large grey box to right of microscope, orange light will glow to indicate the lamp is on. If it does not switch on check the time switch at the wall socket behind the microscope on the left, it may need to be switched on, down position. 3 If you require a transmission image, turn on the halogen lamp power supply to the left of the microscope, black rocker switch, set the brightness a bit over mid range to get started.

4 Select an objective; fitted are Plan 6.3X NA 0.16 Ph1; F 10X NA 0.25; Plan 16X NA 0.35 Ph2; F-LD 32X NA 0.4 and Plan 40X NA 0.6 Ph2 the 16X Ph2 will do for most simple screening jobs 5 Check that the correct phase ring is in the transillumination path i.e. Ph2 for the 16X is the right hand ring.

6 For an epifluorescent image turn off the halogen light source (left box) and push the epifluorescent shutter (rear right) to the central position (open + heat filter), the second (fully in) position is an unfiltered position, you will not see much if any increase in brightness but will do more photo-damage to your sample. 7 Change between wavelengths by pushing/pulling the filter block slider, under the objectives on the left hand side. To keep things simple it has only a Blue/Green (FITC/GFP) and Green/Red (TRITC/Rhodamine) block, there is a UV/Blue (Hoechst/DAPI) block on the halogen lamp power supply, to change loosen the silver knurled screw at the front of the filter block carrier and withdraw the block, insert the UV/Blue block and tighten (gently) the screw, please replace the original filter block when you are done. 8 When you have finished, switch off the Halogen and UV lamp sources and replace the microscope cover, take care not to cover the UV lamp housing on the back if it is still hot.

A general note regarding UV lamps: To minimise the risk of lamp explosions the minimum on and off time for a mercury vapour bulb should be 30mins, please check the lamp housing temperature before switching on if it has obviously recently gone off best practice would be to wait a while before switching back on. It is better to run a lamp for a little longer than required than to repeatedly cycle it. Check the next bulb change due at sticker on the power supply against the timer if it is near or over inform me and I will change it. In the unlikely event of a lamp explosion, leave the room immediately and dont return for at least 30min, inform me and I will clean up the bits.