Visitron Spinning Disk Unit 1

Transcription

1 Startup Procedure and Notes (Last update: Sept. 2014) Switch on the fluorescence illumination for widefield viewing (HXP lamp) 2. Switch on power-bar on the left side of the shelf support 3. Switch on the PC 4. Switch on power-bar on the right side of the shelf support 5. Switch on the stand of the microscope (as soon as the controller of the hardware autofocus displays "Detecting stand") 1 Spinning Disk 1- Startup and Notes MFPL BioOptics - Light Microscopy

2 6. Switch on the laser you will use via the keys 7. Switch on the stage controller/joy stick (at the back side of the unit) 8. Switch on the power bar for the AOTF controller 9. Optional: switch on the power bar for environmental control (water bath, CO 2 and temperature controllers; see below) Shutdown Turn off the lasers (6) followed by the HXP lamp (1). The order of switching off the other components is not important - the usual sequence is (9) Notes: The stand (5) does not need to be switched off. The HXP lamp (1) should cool down for at least 20 min before being turned on again. When you want to use the 405 nm FRAP module, make sure the handle on the left side of the module is pushed in (then no DAPI can be viewed via the eyepiece!). 2 Spinning Disk 1- Startup and Notes MFPL BioOptics - Light Microscopy

3 On screen the movement of x- and y-axis is switched: means when you move in x with the joystick, a y-movement will be displayed on screen and vice versa: the reason is a 90 turned camera chip. When you have run a WF. Macro to get a visual inspection via the eyepieces, make sure to press the "SDC"-button before you start a sequence. Otherwise, the white light from the HXP-lamp will always shine on your sample during acquisition! Environmental Control Water bath for cooled/heated stage insert A B C 9 CO 2 inlet Press the blue button (A, in the center of the controller) and select the desired temperature of the the water bath by turning knob C Press the right blue button (B, "Start") 3 Spinning Disk 1- Startup and Notes MFPL BioOptics - Light Microscopy

4 Press on knob C to start pumping The controller displays two temperature values: the internal temperature of the water bath (T intern ) and the temperature in the insert (T prozess ). Adjust the internal temperature to obtain the desired temperature on stage (T prozess ). Additional insert variants for chambered coverglass slides, ibidi slides and 60 mm dishes are in the box on the shelf behind the computer. This box also contains the screw driver and the screws to fix the inserts (never remove these!). Within the box there is also the 63x objective heater is there inside, which needs to be coupled and controlled via the 37-2 Pecon temp-controller unit. CO2 Controller Check the CO 2 pressure at the pressure reducing valve mounted on the wall behind the microscope (should be approximately 1 bar). Press the "display" button to switch the display between the measured ("real") and set ("nom") CO 2 concentration. Use the "+" or "-" buttons to set the desired concentration. Turn on the flow by pressing the "valve" button. The LED on the button is then either green (flow through the system) or red (no flow). Note that the system does not pump continuously. The LED should turn green in more or less regular intervals. Pressing the "valve" button again turns it off (the LED remains permanently dark). Repeated pressing the "pump" button sets the pump/flow speed (values between 1-5; use 2 or 3). Do not change the knob of the CO 2 reducing valve. 4 Spinning Disk 1- Startup and Notes MFPL BioOptics - Light Microscopy

5 Pecon 37-2 Dual-channel Temperature Controller One channel of the system is used to heat the CO 2 humidifier, the other one can be used to control the 63x objective heater (optional). The display shows the actual temperatures of the channels. Pressing button "1" or "2" (for channel 1 or 2) toggles the display to "Ch 1/2 On/Off" (press "+" to turn a channel on, "-" to turn it off) and the temperature set, respectively (use again the "+" or "-" button to set the value). 5 Spinning Disk 1- Startup and Notes MFPL BioOptics - Light Microscopy