Expand this Topic clickable element to expand a topic
Skip to content
Optica Publishing Group
  • Frontiers in Optics 2009/Laser Science XXV/Fall 2009 OSA Optics & Photonics Technical Digest
  • OSA Technical Digest (CD) (Optica Publishing Group, 2009),
  • paper FThM1
  • https://doi.org/10.1364/FIO.2009.FThM1

Intracellular Nanoscale Imaging with Fluorescence Photoactivation Localization Microscopy

Open Access Open Access

Abstract

Diffraction limits resolution in optical microscopy, but many biological phenomena occur on molecular length scales. Recently, methods to circumvent the diffraction limit have been presented. Fluorescence photoactivation localization microscopy (FPALM) uses multiple iterations of photoactivation, imaging, and photobleaching of subsets of photoactivatable fluorescent probe (PAFP) molecules to generate images with resolution of 10-40 nanometers. The three-dimensional position, brightness, polarization, and other single molecule information are obtained from the raw image(s) of each PAFP. The process is repeated for many subsets to build up data on many molecules. The positions of all localized molecules are used to construct an image of the sample with nanoscale resolution. Results will be shown for a variety of biological systems, including live and fixed cells expressing PAFP fusions of membrane, cytoplasmic, cytoskeletal, and nuclear proteins. Bi-plane FPALM can image in three dimensions, and polarization FPALM can image molecular positions and anisotropies simultaneously. Using these powerful capabilities, many potentially interesting biological problems can be addressed.

© 2009 Optical Society of America

PDF Article
More Like This
Optical Nanoscopy: FPALM Breaks the Diffraction Limit

Travis J. Gould, Manasa V. Gudheti, Mudalige S. Gunewardene, Julie A. Gosse, and Samuel T. Hess
LSWA2 Laser Science (LS) 2009

3D Localization in Fluorescence Photoactivation Localization Microscopy and Particle Tracking

Joerg Bewersdorf, Michael J. Mlodzianoski, Stefanie E. K. Kirschbaum, and Manuel F. Juette
LSThB2 Laser Science (LS) 2009

Development of Novel Fluorescent Proteins for Superresolution Imaging

Tao Xu
NSu1C.1 Nanophotonics, Nanoelectronics and Nanosensor (N3) 2013

Select as filters


Select Topics Cancel
© Copyright 2024 | Optica Publishing Group. All Rights Reserved