Breaking a Dogma: High-Throughput Live-Cell Imaging in Real-Time with Hoechst 33342.
Hoechst 33342
fluorescent microscopy
high-throughput analysis
real-time live-cell imaging
Journal
Advanced healthcare materials
ISSN: 2192-2659
Titre abrégé: Adv Healthc Mater
Pays: Germany
ID NLM: 101581613
Informations de publication
Date de publication:
08 2023
08 2023
Historique:
revised:
14
03
2023
received:
21
01
2023
medline:
9
8
2023
pubmed:
20
3
2023
entrez:
19
3
2023
Statut:
ppublish
Résumé
Automated high-throughput live cell imaging (LCI) enables investigation of substance effects on cells in vitro. Usually, cell number is analyzed by phase-contrast imaging, which is reliable only for a few cell types. Therefore, an accurate cell counting method, such as staining the nuclei with Hoechst 33342 before LCI, will be desirable. However, since the mid-1980s, the dogma exists that Hoechst can only be used for endpoint analyses because of its cytotoxic properties and the potentially phototoxic effects of the excitation light. Since microscopic camera sensitivity has significantly improved, this study investigates whether this dogma is still justified. Therefore, exposure parameters are optimized using a 4× objective, and the minimum required Hoechst concentration is evaluated, allowing LCI at 30-min intervals over 5 days. Remarkably, a Hoechst concentration of only 57 × 10
Identifiants
pubmed: 36934382
doi: 10.1002/adhm.202300230
doi:
Substances chimiques
bisbenzimide ethoxide trihydrochloride
P976261J69
Benzimidazoles
0
Fluorescent Dyes
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e2300230Informations de copyright
© 2023 The Authors. Advanced Healthcare Materials published by Wiley-VCH GmbH.
Références
a) R. Cole, Cell Adhes. Migr. 2014, 8, 452;
b) V. Fetz, H. Prochnow, M. Bronstrup, F. Sasse, Nat. Prod. Rep. 2016, 33, 655.
G. Lämmler, H. R. Schütze, Naturwissenschaften 1969, 56, 286.
I. Hilwig, Z. Zellforsch. Mikrosk. Anat. 1970, 104, 127.
I. Hilwig, A. Gropp, Exp. Cell Res. 1972, 75, 122.
a) H. D. Preisler, Cancer Treat. Rep. 1978, 62, 1393;
b) M. E. Lalande, R. G. Miller, J. Histochem. Cytochem. 1979, 27, 394;
c) M. R. Loken, J. Histochem. Cytochem. 1980, 28, 36;
d) M. R. Loken, Cytometry 1980, 1, 136;
e) G. Szabo, Jr., A. Kiss, S. Damjanovich, Cytometry 1981, 2, 20.
M. J. Lydon, K. D. Keeler, D. B. Thomas, J. Cell. Physiol. 1980, 102, 175.
a) J. Fried, J. Doblin, S. Takamoto, A. Perez, H. Hansen, B. Clarkson, Cytometry 1982, 3, 42;
b) G. Van Zant, C. G. Fry, Cytometry 1983, 4, 40;
c) R. E. Durand, P. L. Olive, J. Histochem. Cytochem. 1982, 30, 111.
a) A. Bernheim, R. Miglierina, Hum. Genet. 1989, 83, 189;
b) E. Erba, P. Ubezio, M. Broggini, M. Ponti, M. D'Incalci, Cytometry 1988, 9, 1;
c) M. Gregoire, D. Hernandez-Verdun, M. Bouteille, Biol. Cell. 1984, 52, 285.
D. Adamski, J. F. Mayol, N. Platet, F. Berger, F. Herodin, D. Wion, FEBS Lett. 2007, 581, 3076.
B. Steuer, B. Breuer, A. Alonso, Exp. Cell Res. 1990, 186, 149.
a) S. S. Wong, R. A. Sturm, J. Michel, X. M. Zhang, P. A. Danoy, K. McGregor, J. J. Jacobs, A. Kaushal, Y. Dong, I. S. Dunn, P. G. Parsons, Biochem. Pharmacol. 1994, 47, 827;
b) C. Wiezorek, Histochemistry 1984, 81, 493;
c) M. Purschke, N. Rubio, K. D. Held, R. W. Redmond, Photochem. Photobiol. Sci. 2010, 9, 1634.
J. Bucevicius, J. Keller-Findeisen, T. Gilat, S. W. Hell, G. Lukinavicius, Chem. Sci. 2019, 10, 1962.
P. P. Laissue, R. A. Alghamdi, P. Tomancak, E. G. Reynaud, H. Shroff, Nat. Methods 2017, 14, 657.
a) S. Lamouille, J. Xu, R. Derynck, Nat. Rev. Mol. Cell Biol. 2014, 15, 178;
b) D. Y. Shu, E. Butcher, M. Saint-Geniez, Int. J. Mol. Sci. 2020, 21, 4271.
H. R. Fuchs, R. Meister, R. Lotke, C. Framme, Exp. Eye Res. 2020, 201, 108258.
a) A. Przekora, T. Zarnowski, G. Ginalska, Cell. Mol. Biol. Lett. 2017, 22, 5;
b) Y. Seko, N. Azuma, Y. Takahashi, H. Makino, T. Morito, T. Muneta, K. Matsumoto, H. Saito, I. Sekiya, A. Umezawa, PLoS One 2008, 3, e3709.
a) Y. T. Cheung, W. K. Lau, M. S. Yu, C. S. Lai, S. C. Yeung, K. F. So, R. C. Chang, Neurotoxicology 2009, 30, 127;
b) K. Jahn, C. Wieltsch, N. Blumer, M. Mehlich, H. Pathak, A. Q. Khan, H. Hildebrandt, H. Frieling, J. Neural Transm. 2017, 124, 1341.
K. C. Wollert, T. Taga, M. Saito, M. Narazaki, T. Kishimoto, C. C. Glembotski, A. B. Vernallis, J. K. Heath, D. Pennica, W. I. Wood, K. R. Chien, J. Biol. Chem. 1996, 271, 9535.
a) H. D. Jampel, Ophthalmology 1992, 99, 1471;
b) G. Ciancio, A. Pollack, M. A. Taupier, N. L. Block, G. L. Irvin, 3rd, J. Histochem. Cytochem. 1988, 36, 1147.
J. Zheng, S. Qi, H. Zhu, X. Xiao, Cell. Mol. Biol. 2016, 62, 27.
X. Zhang, J. Chen, B. Davis, F. Kiechle, Arch. Pathol. Lab. Med. 1999, 123, 921.
M. Athar, N. K. Chaudhury, M. E. Hussain, R. Varshney, Free Radical Res. 2010, 44, 936.