Mondal, Partha P and Curthoys, Nikki M and Hess, Samuel T (2016) Clean localization super-resolution microscopy for 3D biological imaging. In: AIP ADVANCES, 6 (1).
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Abstract
We propose clean localization microscopy (a variant of fPALM) using a molecule filtering technique. Localization imaging involves acquiring a large number of images containing single molecule signatures followed by one-to-one mapping to render a super-resolution image. In principle, this process can be repeated for other z-planes to construct a 3D image. But, single molecules observed from off-focal planes result in false representation of their presence in the focal plane, resulting in incorrect quantification and analysis. We overcome this with a single molecule filtering technique that imposes constraints on the diffraction limited spot size of single molecules in the image plane. Calibration with sub-diffraction size beads puts a natural cutoff on the actual diffraction-limited size of single molecules in the focal plane. This helps in distinguishing beads present in the focal plane from those in the off-focal planes thereby providing an estimate of the single molecules in the focal plane. We study the distribution of actin (labeled with a photoactivatable CAGE 552 dye) in NIH 3T3 mouse fibroblast cells. (C) 2016 Author(s).
| Item Type: | Journal Article |
|---|---|
| Publication: | AIP ADVANCES |
| Publisher: | AMER INST PHYSICS |
| Additional Information: | Copy right for this article belongs to the AMER INST PHYSICS, 1305 WALT WHITMAN RD, STE 300, MELVILLE, NY 11747-4501 USA |
| Department/Centre: | Division of Physical & Mathematical Sciences > Instrumentation Appiled Physics |
| Date Deposited: | 03 Mar 2016 05:24 |
| Last Modified: | 03 Mar 2016 05:24 |
| URI: | http://eprints.iisc.ac.in/id/eprint/53364 |
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