Zeiss LSM 510 Manual

Also see for LSM 510: Quick start

ANNEX LSM 510 DuoScanCarl Zeiss Monitor Diode LSM 510 META DuoScan8-22 B 45-0021 e 03/06The single steps to find the right setting of all the parameters to be set are listed in the following:• Activate the Range Indicator.• Adjustment of Amplifier Offset: the Offset of the PMT channel and diode channel have to fit to eachother to guarantee the best noise reduction.The best way to do the adjustment is the following:• Choose different colors in the Configuration Control window for PMT and diode channel.• Activate Line Scan.• Switch off all laser lines in the Excitation window.• Activate Cont.• Set values for Ampl. Gain to 1 in each channel.• Set the lines visible to the same level as close to the ground level as possible; the values you find forthe Offset in each channel should be negative.A final adjustment of the offset adjustment is done by visually evaluating the noise reduction in the ratioimage. As the Offset value of the PMT channel influences the range setting of the ratio image much lessthan the Offset value of the diode channel, the fine tuning should be done via the PMT offset, ifrequired.As mentioned before, the calculation of the ratio image is very sensitive to different signaloffsets in the two channels used. As the offset is influenced by the scan speed as well as by theAmplifier Gain used, the offset calibration is not valid any more if the scan speed is changed, orthe Ampl. Gain is set to a new value respectively. In most cases a new fine tuning is necessary. Ifthis doesn’t work, the complete calibration process has to be repeated.Another possibility to calibrate the offset values is to set the values to -0.1 as default for both channels,then perform steps 3 and 4 and finally adjust the noise reduction by varying the PMT offset value.If the ratio application is used and the offset has been set to the best reduction of noise in the ratioimage, it is not allowed to change the offset of the PMT channel to change the reduction of backgroundfluorescence, for example. This can be done only if the diode offset is corrected afterwards.Adjustment of Detector GainThe Gain of the PMT should be set with the help of the range indicator function. No ‘red ’ and no ‘blue’pixels should occur in the image of the PMT.