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Impact of deuteration on the ultrafast nonlinear optical response of toluene and nitrobenzene

2019, Karras, Christian, Chemnitz, Mario, Heintzmann, Rainer, Schmidt, Markus A.

Nonlinear pulse propagation inside highly nonlinear media requires accurate knowledge on the temporal response function of the materials used particular in the case of liquids. Here we study the impact of deuteration on the ultrafast dynamics of toluene and nitrobenzene via all optical Kerr gating, showing substantially different electronic and molecular contributions, which was quantified by fitting a multichannel decay model to the data points. Specifically we found that deuteration imposes the time-integrated nonlinearities to reduce particular for toluene which could be caused by both reduced electronic hyperpolarizabilities as well as weaker intermolecular interactions. The results achieved reveal that deuterated organic solvents represent promising materials for infrared photonics since they offer extended infrared transmission compared to their non-deuterated counterparts while maintained strong nonlinear responses.

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Patterned illumination single molecule localization microscopy (piSMLM): user defined blinking regions of interest cellSTORM - Cost-effective Super-Resolution on a Cellphone using dSTORM

2018, Chen, S.-Y., Bestvater, F., Heintzmann, Rainer, Cremer, Christoph

Single molecule localization microscopy (SMLM) has been established as an important super-resolution technique for studying subcellular structures with a resolution down to a lateral scale of 10 nm. Usually samples are illuminated with a Gaussian shaped beam and consequently insufficient irradiance on the periphery of the illuminated region leads to artifacts in the reconstructed image which degrades image quality. We present a newly developed patterned illumination SMLM (piSMLM) to overcome the problem of uneven illumination by computer-generated holography. By utilizing a phase-only spatial light modulator (SLM) in combination with a modified Gerchberg-Saxton algorithm, a user-defined pattern with homogeneous and nearly speckle-free illumination is obtained. Our experimental results show that irradiance 1 to 5 kW/cm2 was achieved by using a laser with an output power of 200 mW in a region of 2000 µm2 to 500 µm2, respectively. Higher irradiance of up to 20 kW/cm2 can be reached by simply reducing the size of the region of interest (ROI). To demonstrate the application of the piSMLM, nuclear structures were imaged based on fluctuation binding-activated localization microscopy (fBALM). The super-resolution fBALM images revealed nuclear structures at a nanometer scale.Single molecule localization microscopy (SMLM) has been established as an important super-resolution technique for studying subcellular structures with a resolution down to a lateral scale of 10 nm. Usually samples are illuminated with a Gaussian shaped beam and consequently insufficient irradiance on the periphery of the illuminated region leads to artifacts in the reconstructed image which degrades image quality. We present a newly developed patterned illumination SMLM (piSMLM) to overcome the problem of uneven illumination by computer-generated holography. By utilizing a phase-only spatial light modulator (SLM) in combination with a modified Gerchberg-Saxton algorithm, a user-defined pattern with homogeneous and nearly speckle-free illumination is obtained. Our experimental results show that irradiance 1 to 5 kW/cm2 was achieved by using a laser with an output power of 200 mW in a region of 2000 µm2 to 500 µm2, respectively. Higher irradiance of up to 20 kW/cm2 can be reached by simply reducing the size of the region of interest (ROI). To demonstrate the application of the piSMLM, nuclear structures were imaged based on fluctuation binding-activated localization microscopy (fBALM). The super-resolution fBALM images revealed nuclear structures at a nanometer scale.

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Automated distinction of shearing and distortion artefacts in structured illumination microscopy

2018, Förster, Ronny, Müller, Walter, Richter, Renè, Heintzmann, Rainer

Any motion during an image acquisition leads to an artefact in the final image. Structured illumination microscopy (SIM) combines several raw images into one high-resolution image and is thus particularly prone to these motion artefacts. Their unpredictable shape cannot easily be distinguished from real high-resolution content. We previously implemented a motion detection specifically for SIM, which had two shortcomings which are solved here. First, the brightness dependency of the motion signal is removed. Second, the empirical threshold of the calculated motion signal was not a threshold at a maximum allowed artefact. Here we investigate which artefacts are still acceptable and which linear movement creates them. Thus, the motion signal is linked with the maximal strength of the expected artefact. A signal-to-noise analysis including classification successfully distinguishes between artefact-free imaging, shearing and distortion artefacts in biological specimens. A shearing, as in wide-field microscopy, is the dominant reconstruction artefact, while distortions arise not until surprisingly fast movements.