Research articles

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  • Improved photoactivatable red fluorescent proteins are generated by including properties desirable for photoactivated localization microscopy (PALM) as selection criteria. The PAmCherry proteins are superior tags for one- and two-color PALM in fixed cells, among other applications. Also in this issue, McKinney et al. present an improved version of the green-to-red EosFP protein.

    • Fedor V Subach
    • George H Patterson
    • Vladislav V Verkhusha
    Article
  • A combination of multiphoton laser scanning microscopy and second harmonic generation (SHG) imaging is used to track tumor associated fibroblasts and extracellular matrix remodeling in living mice. The SHG signal is used for image registration over several days.

    • Jean Y Perentes
    • Trevor D McKee
    • Yves Boucher
    Brief Communication
  • Conventional in situ hybridization protocols lead to loss of microRNAs, which diffuse out of the formaldehyde-fixed sample owing to their small size. Adding a carbodiimide that stably links the microRNA with the protein matrix around it prevents this diffusion and allows detection of miRNAs at very low expression levels.

    • John T G Pena
    • Cherin Sohn-Lee
    • Thomas Tuschl
    Brief Communication
  • S-palmitoylation is a protein post-translational modification involved in trafficking, compartmentalization and membrane-tethering of various proteins. A palmitic acid analog, 17-octadecynoic acid, serves as a metabolically incorporated bioorthogonal click chemistry probe for detecting S-palmitoylated human proteins by mass spectrometry or in-gel fluorescence.

    • Brent R Martin
    • Benjamin F Cravatt
    Brief Communication
  • A device that traps cells in micrometer-sized capture cups efficiently achieves cell pairing and subsequent chemically or electrically induced fusion. The authors show that fused NIH 3T3 cells continue to be viable and morphologically normal off the chip, and they also show reprogramming of mouse embryonic fibroblasts after fusion with embryonic stem cells.

    • Alison M Skelley
    • Oktay Kirak
    • Joel Voldman
    Article
  • A ratiometric fluorescent sensor that reports the ATP/ADP concentration ratio in living cells was created by fusing the bacterial regulatory protein GlnK1 to a circularly permuted fluorescent protein. The sensor detected inhibition of cellular metabolism caused by transient removal of glucose from the cellular medium or administration of a glycolytic inhibitor.

    • Jim Berg
    • Yin Pun Hung
    • Gary Yellen
    Article
  • Pseudorabies viruses encoding fluorescent proteins are a powerful method for mapping neuronal circuits. Now a series of pseudorabies virus strains encoding fluorescent sensors and time-shifted florescent proteins allow dissection of complex circuits with concurrent activity analysis while defining an analysis period during which the neurons are still healthy.

    • Zsolt Boldogkői
    • Kamill Balint
    • Botond Roska
    Brief Communication