Leica SP5 confocal, User Manual

Page 1: ...smbhost microscope users microscope user manuals Leica SP5 Facility Usage Policy 1 You must have the relevant Risk Assessment COSHH form for the work you are undertaking before using imaging facility...

Page 2: ...ting 8 Simultaneous vs Sequential scanning modes 8 Setting up Sequential Scanning Recommended 9 Image Optimisation Procedure 10 Capturing a Multi Wavelength 2D Image Optical Slice 11 Saving Images 11...

Page 3: ...e Laser Emission key to the ON position 3 The PC will boot automatically login using your windows user account If you have been trained on the system a user account will have been created for you 4 La...

Page 4: ...andby 3 Close the LAS AF program and shut down the PC 4 Turn the Laser Emission key to the horizontal position 5 DO NOT SWITCH OFF THE LASER POWER SWITCH UNTIL THE ARGON FAN HAS STOPPED Continue to ne...

Page 5: ...he XY speed of the stage by pressing the black buttons on the left side of the joystick base The black buttons on the right change the speed in Z Finding the sample by eye 1 Choose a fluorescence filt...

Page 6: ...he coverglass 40x lenses can be either a dry or oil type The software menu used to select the lenses tells you if the lens requires oil Configuration Menu Turning the Lasers on 1 Select Configuration...

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Page 8: ...h the width of the slider to the emission spectrum of the dye Avoid overlapping the spectral sliders with the excitation laser lines 5nm min separation 5 Select the Channel colour box for each detecto...

Page 9: ...edure but this time enable the laser line and detector for RFP and switch off the first detector and 488 laser by setting to 0 power 5 Now when you scan the lasers detectors are sequentially turned on...

Page 10: ...each channel a number of times and averaging the resulting signal Only use if you have a noisy image Image capture time is increased as is the likelihood of phoobleaching 7 Rotation allows the whole s...

Page 11: ...re a few blue pixels Press the saturation indicator button again twice to see the channel images in their original colours Capturing a Multi Wavelength 2D Image Optical Slice 1 If you have followed th...

Page 12: ...cus using the joystick Z control until you find the top bottom of your sample Click on the Begin arrow on the diagram to record that position Then move to the opposite end of the sample and click the...

Page 13: ...d box allows you to define how many images should be captured in XY For example type 2x2 for a square grid of 4 tiles 5 Click on the rubbish bin button to remove the scan field setting 6 You can custo...

Page 14: ...itions button will highlight within the stage area the fields of view you have marked for scanning 7 You can delete a single position using the Clear Position button or delete all using the rubbish bi...

Page 15: ...ect the Delete button 5 To save a copy of the image with the scale bar right click on the image then select Snapshot all Exporting Images 1 To export an experiment or single image right click on it in...

Page 16: ...16 FRAP...

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Page 25: ...25 FRET Acceptor Photobleaching...

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Page 32: ...32 FRET Sensitised Emission...

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Page 42: ...42 Spectral Imaging...

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