Simple and efficient delivery of cell-impermeable organic fluorescent probes into live cells for live-cell superresolution imaging

Meng Zhang; Meihua Li; Wenting Zhang; Yubing Han; Yu-Hui Zhang

doi:10.1038/s41377-019-0188-0

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Simple and efficient delivery of cell-impermeable organic fluorescent probes into live cells for live-cell superresolution imaging

Article

- Simple and efficient delivery of cell-impermeable organic fluorescent probes into live cells for live-cell superresolution imaging
- Light: Science & Applications Vol. 8, Issue 4, Pages: 627-637(2019)
- Affiliations：
  
  1.Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan, Hubei 430074, China
  2.MoE Key Laboratory for Biomedical Photonics, School of Engineering Sciences, Huazhong University of Science and Technology, Wuhan, Hubei 430074, China
- Author bio：
  
  Yu-Hui Zhang (zhangyh@mail.hust.edu.cn)
- Funds：
- DOI：10.1038/s41377-019-0188-0
  CLC：
- Published：2019，
  
  Published Online：14 August 2019，
  
  Received：10 December 2018，
  
  Revised：25 July 2019，
  
  Accepted：01 August 2019
- Accepted：
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Zhang, M. et al. Simple and efficient delivery of cell-impermeable organic fluorescent probes into live cells for live-cell superresolution imaging. Light: Science & Applications, 8, 627-637 (2019).
DOI：

Zhang, M. et al. Simple and efficient delivery of cell-impermeable organic fluorescent probes into live cells for live-cell superresolution imaging. Light: Science & Applications, 8, 627-637 (2019). DOI： 10.1038/s41377-019-0188-0.

摘要

Abstract

Numerous commercial organic fluorophores with excellent optical properties are precluded from live-cell superresolution imaging due to poor cell permeability. Here

we develop a simple but effective strategy that renders cells permeable to cell-impermeable

organic fluorescent probes by using a novel peptide vehicle

PV-1. By simple coincubation with PV-1

22 different cell-impermeable

organic fluorescent probes were efficiently delivered into live cells and specifically labeled a variety of organelles. Moreover

PV-1 can simultaneously transfer up to three different probes into live cells. By using PV-1 and these cell-impermeable fluorescent probes

we obtained multicolor

long-term

live-cell superresolution images of various organelles

which allowed us to study the dynamic interactions between them. PV-1

together with these organic fluorescent probes

will greatly broaden the applications of superresolution imaging technology in diverse live-cell studies and opens up a new avenue in the design and application of peptide vehicles.

关键词

Keywords

references

Specht, E. A., Braselmann, E.&Palmer, A. E. A critical and comparative review of fluorescent tools for live-cell imaging.Annu. Rev. Physiol.79, 93–117 (2017)..

Dean, K. M.&Palmer, A. E. Advances in fluorescence labeling strategies for dynamic cellular imaging.Nat. Chem. Biol.10, 512–523 (2014)..

Valm, A. M. et al. Applying systems-level spectral imaging and analysis to reveal the organelle interactome.Nature546, 162–167 (2017)..

Sahl, S. J., Hell, S. W.&Jakobs, S. Fluorescence nanoscopy in cell biology.Nat. Rev. Mol. Cell Biol.18, 685–701 (2017)..

Zhanghao, K. et al. Super-resolution dipole orientation mapping via polarization demodulation.Light.: Sci. Appl.5, e16166 (2016)..

Kner, P. et al. Super-resolution video microscopy of live cells by structured illumination.Nat. Methods6, 339–342 (2009)..

Li, D. et al. Extended-resolution structured illumination imaging of endocytic and cytoskeletal dynamics.Science349, aab3500 (2015)..

Guo, Y. T. et al. Visualizing intracellular organelle and cytoskeletal interactions at nanoscale resolution on millisecond timescales.Cell175, 1430–1442 (2018). E17..

Han, Y. B. et al. Cell-permeable organic fluorescent probes for live-cell long-term super-resolution imaging reveal lysosome-mitochondrion interactions.Nat. Commun.8, 1307 (2017)..

Lukinavičius, G. et al. A near-infrared fluorophore for live-cell super-resolution microscopy of cellular proteins.Nat. Chem.5, 132–139 (2013)..

Lukinavičius, G. et al. Fluorogenic probes for multicolor imaging in living cells.J. Am. Chem. Soc.138, 9365–9368 (2016)..

Panchuk-Voloshina, N. et al. Alexa dyes, a series of new fluorescent dyes that yield exceptionally bright, photostable conjugates.J. Histochem. Cytochem.47, 1179–1188 (1999)..

Mujumdar, R. B. et al. Cyanine dye labeling reagents: sulfoindocyanine succinimidyl esters.Bioconjugate Chem.4, 105–111 (1993)..

Grimm, J. B. et al. A general method to improve fluorophores for live-cell and single-molecule microscopy.Nat. Methods12, 244–250 (2015)..

Jones, S. A. et al. Fast, three-dimensional super-resolution imaging of live cells.Nat. Methods8, 499–505 (2011)..

Hennig, S. et al. Instant live-cell super-resolution imaging of cellular structures by nanoinjection of fluorescent probes.Nano Lett.15, 1374–1381 (2015)..

Erazo-Oliveras, A. et al. Protein delivery into live cells by incubation with an endosomolytic agent.Nat. Methods11, 861–867 (2014)..

Soomets, U. et al. Deletion analogues of transportan.Biochim. et. Biophys. Acta (BBA) - Biomembr.1467, 165–176 (2000)..

Li, W. J., Nicol, F.&Szoka, F. C.Jr. GALA: a designed synthetic pH-responsive amphipathic peptide with applications in drug and gene delivery.Adv. Drug Delivery Rev.56, 967–985 (2004)..

Derossi, D. et al. The third helix of the Antennapedia homeodomain translocates through biological membranes.J. Biol. Chem.269, 10444–10450 (1994)..

Copolovici, D. M. et al. Cell-penetrating peptides: design, synthesis, and applications.ACS Nano8, 1972–1994 (2014)..

Pan, D. et al. A general strategy for developing cell-permeable photo-modulatable organic fluorescent probes for live-cell super-resolution imaging.Nat. Commun.5, 5573 (2014)..

Dubois, J. et al. Fluorescent and biotinylated analogues of docetaxel: synthesis and biological evaluation.Bioorg. Med. Chem.3, 1357–1368 (1995)..

Ma, Q. J. et al. A fluorescent sensor for low pH values based on a covalently immobilized rhodamine–napthalimide conjugate.Sens. Actuators B: Chem.166–167, 68–74 (2012)..

Li, B. et al. A lysosomal probe for monitoring of pH in living cells and ovarian tumour.Dyes Pigments139, 318–325 (2017)..

Georgiev, N. I., Bojinov, V. B.&Venkova, A. I. Design, synthesis and pH sensing properties of novel PAMAM light-harvesting dendrons based on rhodamine 6G and 1, 8-naphthalimide.J. Fluoresc.23, 459–471 (2013)..

Angeles-Boza, A. M. et al. Generation of endosomolytic reagents by branching of cell-penetrating peptides: tools for the delivery of bioactive compounds to live cells in cis or trans.Bioconjugate Chem.21, 2164–2167 (2010)..

Ma, Y. et al. Direct cytosolic delivery of cargoes in vivo by a chimera consisting of D- and L-arginine residues.J. Control. Release162, 286–294 (2012)..

Erazo-Oliveras, A. et al. Improving the endosomal escape of cell-penetrating peptides and their cargos: strategies and challenges.Pharmaceuticals5, 1177–1209 (2012)..

Bucci, C. et al. The small GTPase rab5 functions as a regulatory factor in the early endocytic pathway.Cell70, 715–728 (1992)..

Keppler, A. et al. A general method for the covalent labeling of fusion proteins with small molecules in vivo.Nat. Biotechnol.21, 86–89 (2003)..

Keppler, A. et al. Labeling of fusion proteins ofO6-alkylguanine-DNA alkyltransferase with small molecules in vivo and in vitro.Methods32, 437–444 (2004)..

Keppler, A. et al. Labeling of fusion proteins with synthetic fluorophores in live cells.Proc. Natl Acad. Sci. USA101, 9955–9959 (2004)..

Gautier, A. et al. An engineered protein tag for multiprotein labeling in living cells.Chem. Biol.15, 128–136 (2008)..

Friedman, J. R. et al. ER tubules mark sites of mitochondrial division.Science334, 358–362 (2011)..

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