These technical notes cover hardware related aspects related of our microscope products such as the MicroTime 200 or the LSM Upgrade Kit. This includes, for example, combinations of the MicroTime 200 with AFMs or spectrographs, as well as more detailed descriptions of upgrades of specifc LSMs.
Implementation of a Sample Temperature Control System for the MicroTime 200
The technical note describes the implementation of a sample temperature control system from Bioptechs into the MicroTime 200 for single molecule experiments. It explains the pre-requisites of the microscope setup and technical details of the Bioptechs temperature control system. Raster scanning with the objective heater using objective scanning configuration was tested as well as the temperature accuracy and performance of the complete system. As an application example the temperature dependent RNA folding was studied via FRET. In addition, the temperature dependence on the diffusion coefficient was investigated by two-focus FCS (2fFCS). The appendix explains the main installation steps of the sample temperature controller.
ROI Scanning Using Pulsed Lasers and an Olympus FluoView FV1000 or FluoView FV300
The techical note describes the performance of the PicoQuant pulsed lasers for ROI scanning and bleaching in the Olympus FluoView FV1000. It explains the setup and working principle of the underlying electronic modules and the precise control by the Olympus LSM software laser management system. In addition two typical applications of selective photobleaching and laser ablation are shown.
FLIM: Scanning Speed vs. Excitation Rate
FLIM using TCSPC requires pulsed excitation. This technical note explains the limitations of fast scanning speeds at typical excitation rates for the acquisition of FLIM images. Countermeasures for artifact-free data acquisition like reduced line frequency, increased excitation rate and decreased number of pixels are discussed.