Publications and News

Contents

April 6, 2014

"Fluorescence nanoscopy by polarization modulation and polarization angle narrowing” by Hafi et al Nature Methods, describe a novel approach to optical superresolution imaging that utilizes rotating polarization of the excitation beam. This method is unique among superresolution technologies in that it does not depend on photoswitching or stochastic switching between fluorescence states, but instead singles out individual fluorophores by their relative orientation. The authors demonstrate the technique in cultured neurons using membrane-targeted EGFP — success with this FP suggests that newer, brighter FPs may perform even better with this method.

April 3, 2014

"FALCON: fast and unbiased reconstruction of high-density super-resolution microscopy data” by Min et al., Scientific Reports, describes a novel high-speed algorithm for reconstructing PALM/STORM data. Compared with previous techniques, this algorithm is especially effective at maintaining localization accuracy higher labeling densities. The authors demonstrate very fast (2.5 second resolution) live-cell PALM imaging of endoplasmic reticulum dynamics, opening the door for wider use of localization imaging in live cells.

March 26, 2014

"Nanoscopic spine localization of Norbin, an mGluR5 accessory protein” by Westin et al., BMC Neuroscience, is one recent example of the practical applications of superresolution imaging. The authors used STED (which has lower spatial resolution than PALM/STORM) to clarify the co-localization of several proteins in synapses. They found that Norbin co-localizes with actin but not PSD-95, consistent with co-IP data. Standard confocal microscopy has insufficient resolution to show that Norbin and PSD-95 do not co-localize. This is one of many examples of the power of superresolution imaging to reveal cellular architecture in more detail than has previously been possible.

March 2, 2014

In a recent paper in Nature Photonics, Jia et al. describe a novel approach to increasing localization precision in 3D superresolution imaging using self-bending point spread functions. In this work, they were able to localize single molecules with a precision of 10-15 nm over a depth of 3 μm, and demonstrated the technique for imaging microtubules and mitochondria in fixed cells.

January 13, 2014

"Single-molecule evaluation of fluorescent protein photoactivation efficiency using an in vivo nano template” by Durisic et al.,Nature Methods, presents a comparison of the important photoactivation and photoconversion properties of the most widely used tags in PALM/STORM imaging: mEos2, mEos3.1, mEos3.2, Dendra2, mClavGr2, mMaple, PA-GFP and PA-mCherry. Among the findings reported, both mEos2 and Dendra2 contain subpopulations that are resistant to photoconversion from green to red, and that mEos2 has a high reversible blinking rate that could interfere with imaging at higher densities. Moreover, they found that PA-GFP, the photoactivatable FP with the longest usage history, undergoes a substantial amount of activation with 488 nm illumination, presenting a major practical limitation for PALM imaging.



January 10, 2014

Researchers at IFCO recently described a method using PALM techniques to measure the photoactivation efficiency of various FPs used for superresolution imaging. Their Nature paper was reviewed in GEN.

July 30, 2013

Researchers from EMBL and Max Planck Institute describe the use of stochastic single-molecule superresolution imaging coupled with averaging of thousands of images to elucidate a component of the nuclear pore complex structure down to 1 nm resolution! Check out the article in Science and this commentary in BioTechniques.

July 8, 2013

Check out this new article in BioTechniques describing improved STED imaging.

June 12, 2013

Superresolution imaging is catching on in the popular science media! Check out this new article on superresolution microscopy in ScienceNews

March 24, 2013

Allele is excited to announce the release of our newest fluorescent protein, mNeonGreen, which is a super-performer in superresolution imaging!

mNeonGreen was developed in collaboration with our partner non-profit research institute, the Scintillon Institute.

Shown below to the right is a stochastic single-molecule superresolution (SSMS) image of mNeonGreen fused to the C terminus of human keratin. On the left is the same cell imaged with widefield TIRF microscopy.


Publications

Allele Publications

1. Shaner, N.C. et al. "A bright monomeric green fluorescent protein derived from Branchiostoma lanceolatum." Nature Methods 2013, ePub March 24. Download PDF

This paper describes mNeonGreen and shows several examples of its use in superresolution (SSMS) microscopy.


2. McEvoy A.L. et al. "mMaple: a photoconvertible fluorescent protein for use in multiple imaging modalities." (2012) PLoS One 2012, 7(12):e51314. Download PDF

This paper describes our newest photoconvertible protein, mMaple, and gives examples of its use in different modes of superresolution imaging (STORM and STED).


3. Hoi H. et al. "A monomeric photoconvertible fluorescent protein for imaging dynamic protein localization." J Mol Biol 2010, Sep 3;401(5):776-91. Download PDF

This paper describes the development of mClavGR2, our first photoconvertible protein.

Other Useful References

A comprehensive list of highly useful references for every aspect of superresolution imaging and general microscopy can be found on the Carl Zeiss Microscopy Online Campus.

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