Advances in Confocal Microscopy and Selected Applications

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2021 May 24

PMID 34028709

Citations 7

Authors

W Matt Reilly

Christopher J Obara

Affiliations

Soon will be listed here.

Abstract

Over the last 30 years, confocal microscopy has emerged as a primary tool for biological investigation across many disciplines. The simplicity of use and widespread accessibility of confocal microscopy ensure that it will have a prominent place in biological imaging for many years to come, even with the recent advances in light sheet and field synthesis microscopy. Since these more advanced technologies still require significant expertise to effectively implement and carry through to analysis, confocal microscopy-based approaches still remain the easiest way for biologists with minimal imaging experience to address fundamental questions about how their systems are arranged through space and time. In this review, we discuss a number of advanced applications of confocal microscopy for probing the spatiotemporal dynamics of biological systems.

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References

Waters J . Accuracy and precision in quantitative fluorescence microscopy. J Cell Biol. 2009; 185(7):1135-48. PMC: 2712964. DOI: 10.1083/jcb.200903097. View

Jonkman J, Brown C, Cole R . Quantitative confocal microscopy: beyond a pretty picture. Methods Cell Biol. 2014; 123:113-34. DOI: 10.1016/B978-0-12-420138-5.00007-0. View

Kneen M, Farinas J, Li Y, Verkman A . Green fluorescent protein as a noninvasive intracellular pH indicator. Biophys J. 1998; 74(3):1591-9. PMC: 1299504. DOI: 10.1016/S0006-3495(98)77870-1. View

Frigault M, Lacoste J, Swift J, Brown C . Live-cell microscopy - tips and tools. J Cell Sci. 2009; 122(Pt 6):753-67. DOI: 10.1242/jcs.033837. View

Schnell U, Dijk F, Sjollema K, Giepmans B . Immunolabeling artifacts and the need for live-cell imaging. Nat Methods. 2012; 9(2):152-8. DOI: 10.1038/nmeth.1855. View

Syrbu S, Cohen M . An enhanced antigen-retrieval protocol for immunohistochemical staining of formalin-fixed, paraffin-embedded tissues. Methods Mol Biol. 2011; 717:101-10. DOI: 10.1007/978-1-61779-024-9_6. View

Rodriguez E, Campbell R, Lin J, Lin M, Miyawaki A, Palmer A . The Growing and Glowing Toolbox of Fluorescent and Photoactive Proteins. Trends Biochem Sci. 2016; 42(2):111-129. PMC: 5272834. DOI: 10.1016/j.tibs.2016.09.010. View

Vera M, Tutucci E, Singer R . Imaging Single mRNA Molecules in Mammalian Cells Using an Optimized MS2-MCP System. Methods Mol Biol. 2019; 2038:3-20. PMC: 8950109. DOI: 10.1007/978-1-4939-9674-2_1. View

Trachman 3rd R, Truong L, Ferre-DAmare A . Structural Principles of Fluorescent RNA Aptamers. Trends Pharmacol Sci. 2017; 38(10):928-939. PMC: 7699827. DOI: 10.1016/j.tips.2017.06.007. View

10.

Thorn K . Genetically encoded fluorescent tags. Mol Biol Cell. 2017; 28(7):848-857. PMC: 5385933. DOI: 10.1091/mbc.E16-07-0504. View

11.

Costantini L, Snapp E . Fluorescent proteins in cellular organelles: serious pitfalls and some solutions. DNA Cell Biol. 2013; 32(11):622-7. PMC: 3806368. DOI: 10.1089/dna.2013.2172. View

12.

Lambert T . FPbase: a community-editable fluorescent protein database. Nat Methods. 2019; 16(4):277-278. DOI: 10.1038/s41592-019-0352-8. View

13.

Bukhari H, Muller T . Endogenous Fluorescence Tagging by CRISPR. Trends Cell Biol. 2019; 29(11):912-928. DOI: 10.1016/j.tcb.2019.08.004. View

14.

Ingram J, Schmidt F, Ploegh H . Exploiting Nanobodies' Singular Traits. Annu Rev Immunol. 2018; 36:695-715. DOI: 10.1146/annurev-immunol-042617-053327. View

15.

Asano S, Gao R, Wassie A, Tillberg P, Chen F, Boyden E . Expansion Microscopy: Protocols for Imaging Proteins and RNA in Cells and Tissues. Curr Protoc Cell Biol. 2018; 80(1):e56. PMC: 6158110. DOI: 10.1002/cpcb.56. View

16.

Lavis L . Teaching Old Dyes New Tricks: Biological Probes Built from Fluoresceins and Rhodamines. Annu Rev Biochem. 2017; 86:825-843. DOI: 10.1146/annurev-biochem-061516-044839. View

17.

Grimm J, Klein T, Kopek B, Shtengel G, Hess H, Sauer M . Synthesis of a Far-Red Photoactivatable Silicon-Containing Rhodamine for Super-Resolution Microscopy. Angew Chem Int Ed Engl. 2015; 55(5):1723-7. PMC: 4736676. DOI: 10.1002/anie.201509649. View

18.

Deo C, Sheu S, Seo J, Clapham D, Lavis L . Isomeric Tuning Yields Bright and Targetable Red Ca Indicators. J Am Chem Soc. 2019; 141(35):13734-13738. DOI: 10.1021/jacs.9b06092. View

19.

Deo C, Lavis L . Synthetic and genetically encoded fluorescent neural activity indicators. Curr Opin Neurobiol. 2018; 50:101-108. DOI: 10.1016/j.conb.2018.01.003. View

20.

Martineau M, Somasundaram A, Grimm J, Gruber T, Choquet D, Taraska J . Semisynthetic fluorescent pH sensors for imaging exocytosis and endocytosis. Nat Commun. 2017; 8(1):1412. PMC: 5680258. DOI: 10.1038/s41467-017-01752-5. View