Skip to main content

Microscopy Methods for Imaging MIF and Its Interaction Partners

  • Protocol
  • First Online:
Macrophage Migration Inhibitory Factor

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2080))

Abstract

Fluorescence microscopy has become a powerful tool to investigate proteins in their natural environment. Well-established techniques like widefield and confocal fluorescence microscopy have commonly been used for decades to visualize biomolecules in single cells and tissue sections. Live cell microscopy allows for the investigation of biomolecular trafficking, and other specialized techniques, such as proximity ligation assays (PLA) and fluorescence lifetime imaging microscopy (FLIM), can be used to study interactions between biomolecules of interest. Finally, with the most recent rise of optical super-resolution microscopy, we can investigate target biomolecules in situ with unprecedented detail on the nanometer scale. Here, we discuss various optical microscopy techniques that have successfully been used to image MIF. We highlight applications, advantages, and limitations of each technique. The techniques described here can easily be adapted to investigate other target proteins, their localization, interaction partners, and mechanisms of action.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Combs CA (2010) Fluorescence microscopy: a concise guide to current imaging methods. Curr Protoc Neurosci Chapter 2:Unit2.1. https://doi.org/10.1002/0471142301.ns0201s50

  2. Combs CA, Shroff H (2017) Fluorescence microscopy: a concise guide to current imaging methods. Curr Protoc Neurosci 79:2.1.1–2.1.25. https://doi.org/10.1002/cpns.29

    Article  Google Scholar 

  3. Pawley JB (2006) Handbook of biological confocal microscopy. Springer, New York

    Book  Google Scholar 

  4. Hibbs AR (2004) Confocal microscopy for Biologists. Springer, New York

    Book  Google Scholar 

  5. Calandra T, Roger T (2003) Macrophage migration inhibitory factor: a regulator of innate immunity. Nat Rev Immunol 3(10):791–800. https://doi.org/10.1038/nri1200

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Kim TK, Eberwine JH (2010) Mammalian cell transfection: the present and the future. Anal Bioanal Chem 397(8):3173–3178. https://doi.org/10.1007/s00216-010-3821-6

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Fredriksson S, Gullberg M, Jarvius J, Olsson C, Pietras K, Gústafsdóttir SM, Östman A, Landegren U (2002) Protein detection using proximity-dependent DNA ligation assays. Nat Biotechnol 20:473. https://doi.org/10.1038/nbt0502-473

    Article  CAS  PubMed  Google Scholar 

  8. Olink - Precision Proteomics for Life. https://www.olink.com/

  9. Lang T, Lee JPW, Elgass K, Pinar AA, Tate MD, Aitken EH, Fan H, Creed SJ, Deen NS, Traore DAK (2018) Macrophage migration inhibitory factor is required for NLRP3 inflammasome activation. Nat Commun 9(1):2223. https://doi.org/10.1038/s41467-018-04581-2

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Lakowicz JR (2006) Principles of Fluorescence Spectroscopy. Springer, New York

    Book  Google Scholar 

  11. Becker W (2017) The bh TCSPC handbook. Becker and Hickl GmbH, Berlin

    Google Scholar 

  12. Becker W (2012) Fluorescence lifetime imaging – techniques and applications. J Microsc 247(2):119–136. https://doi.org/10.1111/j.1365-2818.2012.03618.x

    Article  CAS  PubMed  Google Scholar 

  13. Bastiaens PI, Squire A (1999) Fluorescence lifetime imaging microscopy: spatial resolution of biochemical processes in the cell. Trends Cell Biol 9(2):48–52

    Article  CAS  PubMed  Google Scholar 

  14. Bajar BT, Wang ES, Zhang S, Lin MZ, Chu J (2016) A guide to fluorescent protein FRET Pairs. Sensors (Basel, Switzerland) 16(9). https://doi.org/10.3390/s16091488

    Article  PubMed Central  Google Scholar 

  15. Rust MJ, Bates M, Zhuang X (2006) Stochastic optical reconstruction microscopy (STORM) provides sub-diffraction-limit image resolution. Nat Methods 3(10):793–795. https://doi.org/10.1038/nmeth929

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. van de Linde S, Loeschberger A, Klein T, Heidbreder M, Wolter S, Heilemann M, Sauer M (2011) Direct stochastic optical reconstruction microscopy with standard fluorescent probes. Nat Protoc 6(7):991–1009. https://doi.org/10.1038/nprot.2011.336

    Article  CAS  PubMed  Google Scholar 

  17. Hell SW, Wichmann J (1994) Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy. Opt Lett 19(11):780–782. https://doi.org/10.1364/OL.19.000780

    Article  CAS  PubMed  Google Scholar 

  18. Hein B, Willig KI, Hell SW (2008) Stimulated emission depletion (STED) nanoscopy of a fluorescent protein-labeled organelle inside a living cell. Proc Natl Acad Sci U S A 105(38):14271–14276. https://doi.org/10.1073/pnas.0807705105

    Article  PubMed  PubMed Central  Google Scholar 

  19. Sauer M, Heilemann M (2017) Single-molecule localization microscopy in eukaryotes. Chem Rev 117(11):7478–7509. https://doi.org/10.1021/acs.chemrev.6b00667

    Article  CAS  PubMed  Google Scholar 

  20. Allen JR, Ross ST, Davidson MW (2013) Single molecule localization microscopy for superresolution. J Opt 15:094001. https://doi.org/10.1088/2040-8978/15/9/094001

    Article  CAS  Google Scholar 

  21. Gibson TJ, Seiler M, Veitia RA (2013) The transience of transient overexpression. Nat Methods 10:715. https://doi.org/10.1038/nmeth.2534

    Article  CAS  PubMed  Google Scholar 

  22. Allen JR, Ross ST, Davidson MW (2013) Sample preparation for single molecule localization microscopy. Phys Chem Chem Phys 15(43):18771–18783. https://doi.org/10.1039/c3cp53719f

    Article  CAS  PubMed  Google Scholar 

  23. Whelan DR, Bell TD (2015) Image artifacts in single molecule localization microscopy: why optimization of sample preparation protocols matters. Sci Rep 5:7924. https://doi.org/10.1038/srep07924

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Pleiner T, Bates M (2015) Nanobodies: site-specific labeling for super-resolution imaging, rapid epitope-mapping and native protein complex isolation. Elife 4:e11349. https://doi.org/10.7554/eLife.11349

    Article  PubMed  PubMed Central  Google Scholar 

  25. Traenkle B, Rothbauer U (2017) Under the microscope: single-domain antibodies for live-cell imaging and super-resolution microscopy. Front Immunol 8:1030. https://doi.org/10.3389/fimmu.2017.01030

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Schnitzbauer J, Strauss MT, Schlichthaerle T, Schueder F, Jungmann R (2017) Super-resolution microscopy with DNA-PAINT. Nat Protoc 12(6):1198–1228. https://doi.org/10.1038/nprot.2017.024

    Article  CAS  PubMed  Google Scholar 

  27. Nikic-Spiegel I (2018) Genetic code expansion- and click chemistry-based site-specific protein labeling for intracellular DNA-PAINT imaging. Methods Mol Biol 1728:279–295. https://doi.org/10.1007/978-1-4939-7574-7_18

    Article  CAS  PubMed  Google Scholar 

  28. Korczynski J, Wlodarczyk J (2009) Fluorescence lifetime imaging microscopy (FLIM) in biological and medical research. Postepy Biochem 55(4):434–440

    PubMed  Google Scholar 

  29. van Munster EB, Gadella TW (2005) Fluorescence lifetime imaging microscopy (FLIM). Adv Biochem Eng Biotechnol 95:143–175

    PubMed  Google Scholar 

  30. Wheeler A, Henriques R (2017) Standard and super-resolution bioimaging data analysis: a primer. Wiley, Hoboken, NJ

    Book  Google Scholar 

  31. Lee A, Tsekouras K, Calderon C, Bustamante C, Presse S (2017) Unraveling the thousand word picture: an introduction to super-resolution data analysis. Chem Rev 117(11):7276–7330. https://doi.org/10.1021/acs.chemrev.6b00729

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Kirstin D. Elgass .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2020 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Elgass, K.D., Creed, S.J., Rudloff, I. (2020). Microscopy Methods for Imaging MIF and Its Interaction Partners. In: Harris, J., Morand, E. (eds) Macrophage Migration Inhibitory Factor. Methods in Molecular Biology, vol 2080. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9936-1_9

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-9936-1_9

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9935-4

  • Online ISBN: 978-1-4939-9936-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics