Laser Scanning versus Wide-Field—Choosing the Appropriate Microscope in Life Sciences
Source abstract
Abstract
Methods and applications of light microscopy in the life sciences are compared with respect to 3D imaging, resolution, light exposure, sensitivity, and recording time. While conventional wide-field or laser scanning microscopy appear appropriate for smaller samples of only a few micrometers in size with a limited number of light exposures, light sheet microscopy appears to be an optimal method for larger 3D cell cultures, biopsies, or small organisms if multiple exposures or long measuring periods are desired. Super-resolution techniques should be considered in the context of high light exposure possibly causing photobleaching and photo-toxicity to living specimens.
Source: crossref · Quality: baseline
Research context
OpenAlex topics: Advanced Fluorescence Microscopy Techniques · Cell Image Analysis Techniques · Image Processing Techniques and Applications
SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.
Affiliations
Institute of Applied Research, Aalen University, Beethovenstr. 1, 73430 Aalen, Germany
References (73)
Show source-provided references
1. Pawley, J. (1990). Handbook of Biological Confocal Microscopy, Springer. [3rd ed.].10.1007/978-1-4615-7133-9 ↗
2. Confocal optical microscopy10.1088/0034-4885/59/3/003 ↗
3. Method of obtaining optical sectioning by using structured light in a conventional microscope10.1364/OL.22.001905 ↗
4. Single plane illumination module and micro-capillary approach for a wide-field microscope
5. Basic building units and properties of a fluorescence single plane illumination microscope10.1063/1.2428277 ↗
6. Miniaturized modules for light sheet microscopy with low chromatic aberration10.1111/jmi.12439 ↗
7. The mesoSPIM initiative: Open-source light-sheet microscopes for imaging cleared tissue10.1038/s41592-019-0554-0 ↗
8. Preparation strategy and illumination of 3D cell cultures in light–sheet based fluorescence microscopy10.1117/1.JBO.17.10.101518 ↗
9. Imaging intracellular fluorescent proteins at nanometer resolution10.1126/science.1127344 ↗
10. Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM)10.1038/nmeth929 ↗
11. Resolution enhancement techniques in microscopy10.1140/epjh/e2012-20060-1 ↗
12. Heintzmann, R., and Cremer, C. (1999). Laterally modulated excitation microscopy: Improvement of resolution by using a diffraction grating. Optical Biopsies and Microscopic Techniques III, SPIE Press. Proc. SPIE 3568.10.1117/12.336833 ↗
13. Three-dimensional resolution doubling in wide-field fluorescence microscopy by structured illumination10.1529/biophysj.107.120345 ↗
14. Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy10.1046/j.1365-2818.2000.00710.x ↗
15. Simple structured illumination microscope setup with high acquisition speed by using a spatial light modulator10.1364/OE.22.020663 ↗
16. Richter, V., Piper, M., Wagner, M., and Schneckenburger, H. (2019). Increasing Resolution in Live Cell Microscopy by Structured Illumination (SIM). Appl. Sci., 9.10.3390/app9061188 ↗
17. Widefield and total internal reflection fluorescent structured illumination microscopy with scanning galvo mirrors
18. Mechanisms linking obesity to insulin resistance and type 2 diabetes10.1038/nature05482 ↗
19. The many ways to regulate glucose transporter 410.1139/H09-047 ↗
20. Cell-substrate contacts illuminated by total internal reflection fluorescence10.1083/jcb.89.1.141 ↗
21. Total internal reflection fluorescence microscopy: Technical innovations and novel applications10.1016/j.copbio.2004.12.004 ↗
22. Selective imaging of surface fluorescence with very high aperture microscope objectives10.1117/1.1335689 ↗
23. Full-field dual-color 100-nm super-resolution imaging reveals organization and dynamics of mitochondrial and ER networks10.1364/OE.21.026162 ↗
24. A Guide to Structured Illumination TIRF Microscopy at high speed with multiple colors
25. Single-shot super-resolution total internal reflection fluorescence microscopy10.1038/s41592-018-0004-4 ↗
26. Spinning-disk confocal microscopy—A cutting-edge tool for imaging of membrane traffic10.1247/csf.27.349 ↗
27. Schneckenburger, H., Richter, V., and Wagner, M. (2018). Live-Cell Optical Microscopy with Limited Light Doses, SPIE Press.
28. Resolution doubling in fluorescence microscopy with confocal spinning-disk image scanning microscopy10.1073/pnas.1315858110 ↗
29. Image Scanning Microscopy10.1103/PhysRevLett.104.198101 ↗
30. Re-scan confocal microscopy: Scanning twice for better resolution10.1364/BOE.4.002644 ↗
31. Two-photon laser scanning fluorescence microscopy10.1126/science.2321027 ↗
32. Multiphoton microscopy in life sciences
33. A compact STED microscope providing 3D nanoscale resolution10.1111/j.1365-2818.2009.03188.x ↗
34. Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes10.1126/science.aak9913 ↗
35. Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution10.1126/science.1257998 ↗
36. Deep Tissue Imaging with Multiphoton Fluorescence Microscopy10.1016/j.cobme.2017.09.004 ↗
37. Video-rate multi-color structured illumination microscopy with simultaneous real-time reconstruction10.1038/s41467-019-12165-x ↗
38. Spectral imaging and linear unmixing in light microscopy
39. Microspectroscopic fluorescence analysis with prism-based imaging spectrometers: Review and current studies10.1002/cyto.a.20265 ↗
40. Tumor cell differentiation by label-free fluorescence microscopy10.1117/1.JBO.17.10.101508 ↗
41. Multiple frequency fluorescence lifetime imaging microscopy10.1046/j.1365-2818.2000.00651.x ↗
42. Laser-assisted fluorescence microscopy for measuring cell membrane dynamics10.1039/b317047k ↗
43. Time-resolved fluorescence microscopy10.1039/b412924p ↗
44. Modulated CMOS camera for fluorescence lifetime microscopy10.1002/jemt.22587 ↗
45. Fluorescence lifetime imaging—Techniques and applications10.1111/j.1365-2818.2012.03618.x ↗
46. Zwischenmolekulare Energiewanderung und Fluoreszenz10.1002/andp.19484370105 ↗
47. Schneckenburger, H., Weber, P., Wagner, M., Enderle, S., Kalthof, B., Schneider, L., Herzog, C., Weghuber, J., and Lanzerstorfer, P. (2019). Combining TIR and FRET in molecular test systems. Int. J. Mol. Sci., 20.10.3390/ijms20030648 ↗
48. Axial Tomography in Live Cell Laser Microscopy10.1117/1.JBO.22.9.091505 ↗
49. Richter, V., Lanzerstorfer, P., Weghuber, J., and Schneckenburger, H. (2020). Super-resolution live cell microscopy of membrane-proximal fluorophores. Int. J. Mol. Sci., 21.10.3390/ijms21197099 ↗
50. Variable-angle total internal reflection fluorescence microscopy (VA-TIRFM): Realization and application of a compact illumination device10.1046/j.1365-2818.2003.01200.x ↗
51. Nanotopology of cell adhesion upon variable-angle total internal reflection fluorescence microscopy (VA-TIRFM)
52. Multicellular tumor spheroids: A relevant 3D model for the in vitro preclinical investigation of polymer nanomedicines10.1039/C7PY00559H ↗
53. Serial cultivation of normal rat bladder epithelial cells in vitro10.1016/S0022-5347(17)49383-9 ↗
54. Technical bias of microcultivation environments on single cell physiology10.1039/C4LC01270D ↗
55. Monitoring of apoptosis in 3D cell cultures by FRET and light sheet fluorescence microscopy10.3390/ijms16035375 ↗
56. Optical clearing methods: An overview of the techniques used for the imaging of 3D spheroids10.1002/bit.27105 ↗
57. Methods for Optical Skin Clearing in Molecular Optical Imaging in Dermatology
58. Advanced CUBIC protocols for whole-brain and whole-body clearing and imaging10.1038/nprot.2015.085 ↗
59. In-Vivo and ex-vivo optical clearing methods for biological tissues: Review10.1364/BOE.10.005251 ↗
60. Expansion Microscopy: Principles and Uses in Biological Research10.1038/s41592-018-0219-4 ↗
61. Optical projection tomography10.1146/annurev.bioeng.6.040803.140210 ↗
62. Sample holder for axial rotation of specimens in 3D Microscopy10.1111/jmi.12263 ↗
63. Selective plane illumination microscopy techniques in developmental biology10.1242/dev.022426 ↗
64. Light-sheet imaging of mammalian development10.1016/j.semcdb.2015.11.001 ↗
65. Long-term fluorescence live imaging of Tribolium castaneum embryos: Principles, resources, scientific challenges and the comparative approach10.1016/j.cois.2016.08.002 ↗
66. A guide to light-sheet fluorescence microscopy for multiscale imaging10.1038/nmeth.4224 ↗
67. Fluorescence techniques in developmental biology10.1007/s12038-018-9768-z ↗
68. Single-molecule super-resolution light-sheet microscopy10.1002/cphc.201300732 ↗
69. Light sheet microscopy for real-time developmental biology10.1016/j.gde.2011.09.009 ↗
70. Sample Preparation and Mounting of Drosophila Embryos for Multiview Light Sheet Microscopy10.1007/978-1-4939-6371-3_10 ↗
71. High resolution deep view microscopy of cells and tissues10.1070/QEL17204 ↗
72. Photoswitching-Enabled Contrast Enhancement in Light Sheet Fluorescence Microscopy10.1021/acsphotonics.6b00838 ↗
73. Improving image contrast in fluorescence microscopy with nanostructured substrates10.1364/OE.23.029772 ↗