The multiplexing of fluorescence correlation spectroscopy (FCS), especially in imaging FCS using fast, sensitive array detectors, requires the handling of large amounts of data. One can easily collect in excess of 100,000 FCS curves a day, too many to be treated manually. Therefore, ImFCS, an open-source software which relies on standard image files was developed and provides a wide range of options for the calculation of spatial and temporal auto- and cross-correlations, as well as differences in Cross-Correlation Functions (ΔCCF). ImFCS permits fitting of standard models to correlation functions and provides optimized histograms of fitted parameters. Applications include the measurement of diffusion and flow with Imaging Total Internal Reflection FCS (ITIR-FCS) and Single Plane Illumination Microscopy FCS (SPIM-FCS) in biologically relevant samples. As a compromise between ITIR-FCS and SPIM-FCS, we extend the applications to Imaging Variable Angle-FCS (IVA-FCS) where sub-critical oblique illumination provides sample sectioning close to the cover slide.

Objectives A user interface for transfer function (TF) definition in volume rendering (VR) was developed that allows the user to intuitively assign color and opacity to the original image intensities. This software may surpass solutions currently deployed in clinical practice by simplifying the use of TFs beyond predefined settings that are not always applicable. Materials and methodsThe TF definition is usually a cumbersome task that requires the user to manipulate graphical representations of the TF (e.g. trapezoids). A new method that allows the user to place markers at points of interest directly on CT and MRI images or orthogonal reformations was developed based on two-dimensional region growing and a few user-definable marker-related parameters. For each user defined image marker, a segment of the transfer function is computed. The resulting TF can also be applied to the slice image views. Results were judged subjectively. Results Each individualized TF can be defined interactively in a few simple steps. For every user interaction, immediate visual feedback is given. Clinicians who tested the application appreciated being able to directly work on familiar slice images to generate the desired 3D views. M. Teistler (B) · W. L. NowinskiConclusion Interactive TF definition can increase the actual utility of VR, help to understand the role of the TF with its variations, and increase the acceptance of VR as a clinical tool.