Correlative light microscopy for high-content screening

Biotechniques. 2013 Nov;55(5):243-52. doi: 10.2144/000114099.

Abstract

High-throughput microscopy is an effective tool for rapidly collecting data on a large scale. However, high throughput comes at the cost of low spatial resolution. Here we introduce correlative light microscopy by combining fast automated widefield imaging, confocal microscopy and super-resolution microscopy. We demonstrate the potential of this approach for scalable experiments. The workflow consists of a robust approach for selecting cells of interest on a wide-field screening microscope at low resolution and subsequently re-localizing those cells with micrometer precision for confocal and super-resolution imaging. As a case study, we visualized and quantified cis- and trans-Golgi markers at increasing resolution.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Golgi Apparatus / ultrastructure*
  • Image Processing, Computer-Assisted / methods
  • Kidney / cytology
  • Microscopy, Confocal / methods*
  • Microscopy, Fluorescence / methods*
  • Rats