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Utilisation of Toluidine blue O pH 4.0 and histochemical inferences in plant sections obtained by free-hand

  • New Methods in Cell Biology
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Abstract

For the observation of plant material under a light microscope, free-hand sectioning is usually the fastest and most practical procedure. The staining of the obtained cuts is a crucial step and the action mechanism of the dye is variable. Toluidine blue O is a cationic dye whose metachromatic potential for free-hand sections has long been known. Thus, this work aimed at analysing the performance of Toluidine blue O for this type of sectioning with a systematic and comparative approach. The action of Toluidine blue O pH 4.0 was tested for both fresh and previously clarified free-hand sections from leaves of different species of plant. These sections were compared with sections embedded in glycol-methacrylate resin stained with the same dye or submitted to different histochemical tests. The fresh sections obtained by free-hand and stained with Toluidine blue O permitted the accomplishment of histochemical inferences, with the advantage of maintaining the natural colours from the tissues. The previously clarified sections presented the advantage of having a clearer image, allowing a better cellular delimitation. We concluded that Toluidine blue O is very informative for sections of plant material obtained by free-hand, being, therefore, a procedure that is fast, inexpensive and with excellent results.

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Acknowledgements

We thank Prof. PhD Claudenir Simões Caires, from the Department of Natural Sciences, and Prof. PhD Dalton Longue Júnior from the Department of Phytotechny and Zootechny of the Universidade Estadual do Sudoeste da Bahia for the important suggestions in the manuscript writing; and the Universidade Estadual do Sudoeste da Bahia for granting to Valdir Carvalho Ribeiro the opportunity to work as a voluntary intern in the Botany Laboratory and for all the material provided during the course of this research.

Funding

This work was supported by a voluntary internship granted to Valdir Carvalho Ribeiro by the Universidade Estadual do Sudoeste da Bahia Botany Laboratory.

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Authors and Affiliations

  1. Undergraduate Student of Biological Sciences, Universidade Estadual do Sudoeste da Bahia - UESB, Vitória da Conquista, Bahia State, 45031-900, Brazil

    Valdir Carvalho Ribeiro

  2. Departamento de Ciências Naturais, Universidade Estadual do Sudoeste da Bahia - UESB, Vitória da Conquista, Bahia State, 45031-900, Brazil

    Carlos André Espolador Leitão

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  1. Valdir Carvalho Ribeiro
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  2. Carlos André Espolador Leitão
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Contributions

Valdir Carvalho Ribeiro assisted in the preparation of the slide collection, the analyses of the results, the manuscript writing and translation. Carlos André Espolador Leitão was the supervisor of the work and the one responsible for the idealisation of using Toluidine blue O (TBO) staining as a technique for free-hand sections previously clarified with sodium hypochlorite. He also performed all laboratory procedures and wrote the manuscript together with Valdir Carvalho Ribeiro and provided the needed equipment and reagents.

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Correspondence to Carlos André Espolador Leitão.

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Ribeiro, V.C., Leitão, C.A.E. Utilisation of Toluidine blue O pH 4.0 and histochemical inferences in plant sections obtained by free-hand. Protoplasma 257, 993–1008 (2020). https://doi.org/10.1007/s00709-019-01473-0

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