Chapter 5: Imaging in depth: controversies and opportunities

Methods in Cell Biology
Don O'Malley

Abstract

One enduring challenge of biological imaging is achieving depth of penetration-into cells, tissues, and animals. How deeply can we probe and with what resolution and efficacy? These are critical issues as microscopists seek to push ever deeper, while resolving structural details and observing specific molecular events. In this guide to depth-appropriate modalities, standard optical platforms such as confocal and two-photon microscopes are considered along with complementary imaging modalities that range in depth of penetration. After an introduction to basic techniques, the trade-offs and limitations that distinguish competing technologies are considered, with emphasis on the visualization of subcellular structures and dynamic events. Not surprisingly, there are differences of opinion regarding imaging technologies, as highlighted in a section on point-scanning and Nipkow-disk style confocal microscopes. Confocal microscopy is then contrasted with deconvolution and multi-photon imaging modalities. It is also important to consider the detectors used by current instruments (such as PMTs and CCD cameras). Ultimately specimen properties, in conjunction with instrumentation, determine the depth at which subcellular operations and la...Continue Reading

Citations

Mar 23, 2011·Journal of Biophotonics·Viera CrosignaniEnrico Gratton
Mar 19, 2016·Chemistry, an Asian Journal·Divya Susan PhilipsAyyappanpillai Ajayaghosh

❮ Previous
Next ❯

Related Concepts

Related Papers

Microscopy Research and Technique
John M Girkin, G McConnell
Current Opinion in Microbiology
D C Fung, J A Theriot
Nature Photonics
Erich E Hoover, Jeff A Squier
© 2021 Meta ULC. All rights reserved