![]() ![]() The automated microscope was then programmed to collect a z-stack of 32 images ( z = 0 at the optimal focal plane, 16 images above the focal plane, 16 below) with 2 µm between slices. For each site, the optimal focus was found using laser autofocusing on the Hoechst channel. The 384-well plates containing U2OS cells stained with Hoechst 33342 and Alexa Fluor 594 phalloidin markers were imaged with an exposure of ms for Hoechst and phalloidin, respectively, at 20× magnification and 2× binning. HCS microscopy images were acquired using an ImageXpress Micro automated cellular imaging system (Molecular Devices, Sunnyvale, CA). Solutions are needed that package together data metrics and visualizations in a user-friendly software tool compatible with any automated microscope, to enable automatic detection of image aberrations that would otherwise corrupt the data and diminish the experimental results. ![]() Commercial automated systems do not package QC metrics and visualizations as a cohesive toolset readily accessible to users researchers may resort to custom software solutions 1 or to visual exploration of hits to exclude obvious artifacts. Analysis of these data sets requires the researcher to explore and visualize gigabytes of image and extracted feature information simple spreadsheet programs (e.g., Microsoft Excel) are inadequate for the task. Anomalies in image quality can pollute high-content microscopy data and in some cases even preclude certain types of experiments. However, these advances have also generated the need for robust metrics and workflows for quality control (QC) of HCS images, typically to reduce the number of images that are spuriously scored as screening hits. All subjects Allied Health Cardiology & Cardiovascular Medicine Dentistry Emergency Medicine & Critical Care Endocrinology & Metabolism Environmental Science General Medicine Geriatrics Infectious Diseases Medico-legal Neurology Nursing Nutrition Obstetrics & Gynecology Oncology Orthopaedics & Sports Medicine Otolaryngology Palliative Medicine & Chronic Care Pediatrics Pharmacology & Toxicology Psychiatry & Psychology Public Health Pulmonary & Respiratory Medicine Radiology Research Methods & Evaluation Rheumatology Surgery Tropical Medicine Veterinary Medicine Cell Biology Clinical Biochemistry Environmental Science Life Sciences Neuroscience Pharmacology & Toxicology Biomedical Engineering Engineering & Computing Environmental Engineering Materials Science Anthropology & Archaeology Communication & Media Studies Criminology & Criminal Justice Cultural Studies Economics & Development Education Environmental Studies Ethnic Studies Family Studies Gender Studies Geography Gerontology & Aging Group Studies History Information Science Interpersonal Violence Language & Linguistics Law Management & Organization Studies Marketing & Hospitality Music Peace Studies & Conflict Resolution Philosophy Politics & International Relations Psychoanalysis Psychology & Counseling Public Administration Regional Studies Religion Research Methods & Evaluation Science & Society Studies Social Work & Social Policy Sociology Special Education Urban Studies & Planning BROWSE JOURNALS
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |