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MICROSCOPY 87


sample, catching the fastest responses. Synchronising excitation and imaging with this SIM scanner setup is ideal for quantitative photomanipulation experiments and also enables advanced optogenetics studies.


Tis neuromodulation technique controls and monitors neuronal activity using light sensitive proteins channelrhodopsin and halorhodopsin, and with the FVMPE-RS, laser light stimulation of these optogenetic ‘switches’ is achieved alongside simultaneous real-time imaging of neuronal cell activity.


Also of importance in many biological processes are the interactions of cells within tissues or organs. Measuring rapid fluctuations in groups of cells with a high signal-to- noise ratio is now achieved with both precision and speed using multipoint mapping, an approach where dedicated scanning patterns allow measurements of fluorescence and electrical responses without crosstalk from neighbouring pixels – ideal for electrophysiology, optogenetics or studies with a systems biology focus.


For example, this capability can generate information into how specific cell types contribute to the overall network function of an organ such as the brain.


Precise multicolour excitation and imaging Providing further insights into cell connectivity and how different cellular structures interact, the FVMPE-RS is optimised for tracking multiple molecular species simultaneously.


Trough multi-wavelength excitation, crosstalk is minimised to produce a clearer definition between multiple fluorophores. Moreover, with the new system, Olympus has introduced a four- axis auto-alignment system for


precise alignment of multiple laser beams, eliminating pixel shifts caused by excitation beam angle mismatches. Tis results in clear separation of multiple fluorophores, enabling multiphoton excitation multicolour experiments of unmatched quality.


Deep-tissue observation has revolutionised many areas of life science, for example enabling in vivo studies of brain function – reaching depths of up to 1.3mm under in vivo conditions.


Depths of 8mm can also be reached in non-living tissue treated with the Scaleview tissue transparency reagent or other methods (Fig. 1.) shows visualisation of neuronal plasticity at a depth of 2mm). Such investigations have been made possible not only by the deep penetration of IR light through tissue, but also by improvements in detection sensitivity.


From dedicated multiphoton objectives to a high-sensitivity GaAsP detector, optical efficiency is prioritised at every point of the FVMPE-RS’ light path, achieving bright, high- resolution images deep within the sample. Allowing the use of


low laser power protects living cells against phototoxicity, and precisely adapting the laser to match demanding sample conditions is also realised in ‘Deep Focus Mode’, which is ideal for in vivo samples with heavy scattering.


Expanding the IR range to accommodate a wider range of fluorophores, the FVMPE-RS now offers optimal multiphoton excitation up to 1300nm, with the ability to support wavelengths of 1600nm. Tese longer wavelengths are also well suited to studies utilising third- and second-harmonic generation techniques (Fig. 2). Such label- free imaging allows researchers to visualise structures such as collagen and haemoglobin in their natural state – a powerful technique when combined with in vivo imaging.


Uniting speed, precision and sensitivity with an extended IR range, the FVMPE-RS system is ideally suited to a variety of deep- observation, in vivo imaging applications.


For more information ✔ at www.scientistlive.com/eurolab


Bülent Peker is product manager of Laser Scanning Microscopy, Olympus Europa. www.olympus-europa.com/microscopy


www.scientistlive.com


Fig. 2. Tail muscles of a zebra fish visualised with the FVMPE-RS via second harmonic generation (SHG; green).


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