Page 7 - Live-cellanalysis handbook

From Images to Answers

Introduction

The nature of cell biology research typically requires that image- At the core of all of these manipulations are numbers – images are
based methods are used to capture moments in time to enable comprised of pixels (picture elements), and each pixel in an image
comparisons between treatment groups and across imaging has a digital value representing the brightness or intensity of that
modalities. Sample information is typically acquired using a portion of the sample, at a specific moment in time. By operating
microscope and a digital camera, and those moments in time are on these values, either in isolation, or while considering nearby
processed and analyzed. Images captured with a typical microscope values, the information in the images can be cleaned of aberrant
camera are digital representations of the analog information information, and data relevant to the imaged sample can be
contained in the sample, providing a means to automatically analyze extracted and measured.
the information in the sample. Once these digital snapshots are
acquired, image processing is used to clean up the data, and image
analysis is used to extract usable information for analysis.

The Image Processing Workflow

Visualization and • Assess data.
Sample Image
Preprocessing • Correct image defects.
• Bleaching.

Restoration and • Restore useful information.
Reconstruction • Kernel filtering.
User driven
or Specify Features • Suppress noise.
automated • Identify regions of interest.

Analysis • Extract parameters like area,
overlap, object number.

Classification • Group objects into
different classes.
• Display population data.
Image processing and analysis is accomplished using a number of techniques, guided by expert knowledge and
software guidance. To ensure processing consistency across static and kinetic data, it is important to establish
a set of image processing parameters which enable operation on all images in an identical manner. This
contextually derived data processing workflow will seamlessly and automatically perform all of the necessary
pre- and post- image processing steps, up to and including object analysis and graphical representation of the
experimental result. Properly designed image analysis workflows are intended to require no human intervention
and processes image archives, generating consistent and actionable results either in real-time, or post-acquisition.

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