TECHNICAL NOTE Speed: Opera Phenix Plus high-content screening system technical performance. As shown by an analysis of HCS-related publications, the number of high-content screens being run each year continues to increase, however a large number of screens are not as “high-content” as they could be (Singh et al. 2014). The majority of high-content screens described in published papers rely on only 1-2 image-based features rather than multi-parametric results for single cells. This is indicative of more target-based approaches being used in drug screening. In very recent times, there is evidence of a reversal towards more phenotypic based screens. Prior to this, reasons for the limited adoption of phenotypic screening approaches might have included insufficient hardware but also the lack of suitable software to analyze complex phenotypes (see also Roberts and Tesdorpf 2015). The Opera Phenix® Plus high-content screening system has been specifically developed to enable users to perform highly multi-parametric phenotypic screens effectively. Authors Angelika Foitzik Ruben Prange Karin Böttcher Alexander Schreiner Revvity, Inc. Waltham, MA The Opera Phenix Plus combines state of the art hardware to allow simultaneous acquisition of up to four fluorescent channels, with Harmony® high-content imaging and analysis software for convenient experimental set-up and analysis. The following table summarizes the new features and main improvements of the Opera Phenix Plus system and explains how they benefit acquisition and analysis speed. For research use only. Not for use in diagnostic proced

Speed: Opera Phenix Plus high-content screening system technical performance. Table 1. Technical advancements of the Opera Phenix Plus high-content screening system compared to one of its predecessors, the Opera QEHS system

Speed: Opera Phenix Plus high-content screening system technical performance. for exceptional image quality at high speed. In particular, efficiency. This leads to shorter exposure times, higher the high numerical aperture of water immersion objectives resolution, less photo bleaching and improved 3D imaging. increases the acquisition speed dramatically. Using a water A further increase in acquisition speed can be achieved by immersion objective lens instead of an air lens reduces the exposure time about 5-fold, due to its better light collecting Figure 1.Speed improvement in a crosstalk-sensitive...

Speed: Opera Phenix Plus high-content screening system technical performance. exposure time per channel, one field per well and confocal Hence, the acquisition time for a given plate type is the mode (Synchrony Optics active). same on a four camera system, regardless of the number of The different plates were measured either with one, two or four camera systems to show the power of Synchrony Optics in combination with multiple cameras. Measured acquisition times for all three plate types are summarized in Fig. 3. The acquisition time for a single channel is the same on all systems. However, whenever...

Acquisition Time in Minutes Speed: Opera Phenix Plus high-content screening system technical performance. Figure 3. Summary of acquisition times for different plate types on 1-, 2- or 4-camera systems. The time in min to record a complete plate on a 1-, 2- and 4-camera system with different numbers of channels (1C = one channel, 2C = two channels, 3C = three channels, 4C = four channels) under standardized conditions (100 ms exposure time per channel, 1 center field per well, 20x water immersion lens, BIN2, confocal acquisition mode) is shown. On a 4-camera system the acquisition time for one...

Speed: Opera Phenix Plus high-content screening system technical performance. conditions, the Opera Phenix Plus data set was analyzed in contains a third of the number of images. This requires less a third of the time. This can be explained by the larger field overhead for loading and decompressing of images during of view of the sCMOS camera, leading to a data set that only Acapella is the high-content imaging and analysis software for the Opera QEHS high-content screening system, which is no longer commercially available. * Figure 4. Opera Phenix Plus delivers results faster due to improved...

Speed: Opera Phenix Plus high-content screening system technical performance. The Opera Phenix Plus high-content screening system is Shantanu Singh, Anne E. Carpenter, Auguste Genovesio: specifically designed to tackle even the most demanding Increasing the Content of High-Content Screening, J Biomol assays, whether the demanding part of the campaign results from the image acquisition side or the analysis side. The system can acquire four channel image sets from 100,000 wells per day and even larger number of images when running z-stacks, which easily adds up to several terabytes of data depending...