Leica SP5C Manual

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[Page 1] Leica SP5C

_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual CALM – Confocal and Advanced Light Microscopy Facility Queen’s Medical Research Institute – College fo...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual Contents 1. Starting up the SP5..................................................................................

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 1 1. Starting up the SP5 1.1. Before you switch on the system, fill in the log book (start time, lasers used, hour...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 2 1.7. The software will start the initialisation of the microscope stand and the scan head, and it cannot be use...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 3 Important notes regarding the use of objective lenses: It is vital for the longevity of the lenses that the co...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 4 2.2. If required, switch on the mercury vapour arc lamp (for detailed user guide see chapter 3. below). Gently ...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 5 Fig. 6 Left side of DMI6000 stand 2.7. Light intensity adjustment In both modes the illumination intensity...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 6 Fig. 7 Front panel of the microscope stand 2.9. To focus on your sample, you can use the two main knobs on...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 7 3.3. Before switching off the lamp at the end of your session, check the booking system to find out whether anot...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 8 Fig. 9 Right hand side of the microscope stand All settings of the DMI6000 microscope stand are shown o...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 9 Open the ‘Configuration’ tab and click the ‘Laser’ icon to open the appropriate window (see Fig. 11). Swit...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 10 Fig. 12 Configuration tab – settings for the USB control panel 4.3. Defining the ‘Beam Path Settings’...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 11 The example in Fig. 13 shows the excitation (dotted lines) and emission spectra (solid lines) of 3 different fl...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 12 4.4. Generating new Beam Path settings The Beam Path window (in the acquisition tab) is a schematic representati...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 13 Fig. 15 Spectroscopic detection of a system with 5 PMTs Fig. 16 Slit and PMT settings control w...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 14 Thumb rules for adjusting the slits: The slits should be set so they do not overlap with any of the laser lines....

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 15 Fig. 18 Control console Fig. 19 display mode, pixels that are close to saturation and over appear ...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 16 4.4.6. Additional transmitted light channel In parallel to acquiring different spectral channels via PMT 1 to 3...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 17 The image acquisition in transmitted light mode requires the use of the 488 nm Argon laser line and can be reco...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 18 If you want to reuse a certain preset at a later time, pull out the drop-down list (Fig. 25 B) and choose from t...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 19 In the ‘Sequential scan’ window activate scan 1 by clicking the appropriate button, which will turn red (see ...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 20 Fig. 26 Main operation buttons If you are ready for recording simply press the ‘Start’ button (see Fi...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 21 Fig. 28 Scan parameters 4.7.2. Pinhole size The pinhole size is given in Airy units (AU) and the diamete...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 22 dimensions of the entire image. The approximate XY-pixel dimensions at maximum resolution using a sampling rate t...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 23 To programme a Z-stack acquisition, pull out the ‘Z-Stack’ panel in the ‘Acquisition’ window on the left ...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 24 it by pressing the arrow next to the ‘End’ position display field. Now you have defined the Z-stack and the s...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 25 bottom-right corner of the frame (see Fig. 33). The system now will only scan and display the area (ROI) defined ...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 26 Fig. 33 ROI selection tool 5. Saving and exporting your data files It is very important to know that a...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 27 This name contains the date in a format that allows the computer to always put it in the right chronological or...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 28 without backing them up. If you have not saved your files properly, you might loose large parts of your work. ...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 29 Log the number of hours you have used the mercury lamp together with the switch off time on the log sheet. State ...

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_____________________________________________________________________________________ Rolly Wiegand – CALM Leica SP5 manual 30 Check whether the laser unit is turned on (switch and key) on the front panel (see Fig. 1). Confirm that the la...