Image deconvolution algorithms reverse out-of-focus fluorescent blurring in widefield microscopy
Widefield microscopy is a powerful tool that has been utilized to investigate cellular and molecular processes involved in cell signaling. Although fluorescent blurring from out-of-focus light is an innate result from all widefield microscopes, image deconvolution algorithms can reverse this out-of-focus light artifact.
The Digital Confocal Option is a 2D real-time deconvolution technology that helps to restore a cellular image’s resolution and contrast by mathematical deblurring of the image through reversal of the point spread function. As a result, images and fine detail become sharper while maintaining quantitative information.
Significantly increase throughput while maintaining assay quality
The Digital Confocal Option* enhances contrast of images during acquisition, allowing you to decrease exposure time more than two fold and increase throughput while maintaining assay quality. Alternatively, assays with low Z’-factors can be improved more than 25%. This intelligent analysis enables researchers to increase their throughput, allowing acquisition and analysis of more than 10 million cells per hour.
The speed and convenience offered by the new module delivers a real advantage for scientists as it provides the user with real-time deconvolution without the need to post-process the images. For researchers working in high-content screening, significant resources are dedicated to screening millions of samples for rare new drug or gene therapy candidates.
Digital Confocal Option is designed for the ImageXpress Pico and ImageXpress Micro system solutions.
Designed for the ImageXpress Pico System and CellReporterXpress Image Acquisition and Analysis Software
Significantly increase resolution and assay quality with Digital Confocal 2D on-the-fly deconvolution. The Digital Confocal Option* allows you to decrease exposure time while maintaining statistical significance of your observations. Digital Confocal is seamlessly integrated into the ImageXpress Pico system's fluorescent image acquisition workflow allowing you to capture images with higher signal-to-noise data for more precise segmentation and analysis.
*ImageXpress Pico Digital Confocal Option uses AutoQuant 2D Real Time Deconvolution.
Without digital confocal
With digital confocal
Designed for the ImageXpress Micro Systems and MetaXpress High Content Image Acquisition and Analysis Software
The new image deconvolution module delivers a real advantage for scientists working in high-content screening, where significant resource is dedicated to screening millions of samples for rare new drug or gene therapy candidates. The integration of ImageXpress Micro systems with the Digital Confocal Option for MetaXpress software provides unrivaled performance for ease-of-use, while providing high speed and quality images.
Digital Confocal Option enables robust image analysis
Images with Digital Confocal Option enabled during acquisition (bottom) show cell structures more clearly than without (top), enabling more robust image analysis.
Digital Confocal Option sharpens intensity peaks of fluorescent objects
Images of HEK 293 cells containing paraspeckles were acquired with the ImageXpress Micro XL System without (top) and with Digital Confocal Option (K value 0.063) (bottom). The histograms of a line scan through representative images shows how the Digital Confocal Option can sharpen the intensity peaks of the fluorescent objects. A more accurate count of the paraspeckles was achieved when analyzing the digital confocal images with the Granularity Application Module for MetaXpress software (right).
Applications notes
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Improve nuclear translocation assay results using image deconvolution
Assays requiring precise identification of sub-cellular structures or molecules within different compartments of the cell can benefit from the ability of on-the-fly image deconvolution to improve identification of the specific fluorescence signal against potentially high background caused by out-of-focus light. We utilized the ImageXpress Pico system and CellReporterXpress software to assess the translocation of NF-κB from the cytoplasm to the nucleus of HeLa cells upon treatment with TNF-α.
Improve nuclear translocation assay results using image deconvolution
Assays requiring precise identification of sub-cellular structures or molecules within different compartments of the cell can benefit from the ability of on-the-fly image deconvolution to improve identification of the specific fluorescence signal against potentially high background caused by out-of-focus light. We utilized the ImageXpress Pico system and CellReporterXpress software to assess the translocation of NF-κB from the cytoplasm to the nucleus of HeLa cells upon treatment with TNF-α.
High-throughput imaging assays using zebrafish
The ImageXpress® system provides optimal flexibility for acquiring high quality images with a large field of view. MetaXpress® software enables image analysis for a wide range of applications with simple workflows, and in combination with AcuityXpress informatics software for data mining, this end-to-end solution dramatically improves throughput for zebrafish-based in vivo screening.
High-throughput imaging assays using zebrafish
The ImageXpress® system provides optimal flexibility for acquiring high quality images with a large field of view. MetaXpress® software enables image analysis for a wide range of applications with simple workflows, and in combination with AcuityXpress informatics software for data mining, this end-to-end solution dramatically improves throughput for zebrafish-based in vivo screening.
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