A simple method of plant sectioning using the agarose embedding technique for screening intracellular green fluorescent protein

It is difficult to observe plant tissue sections transformed using the agroinfiltration method under a fluorescent microscope. This is due to the softness of the post‐transformation plant. This research was conducted to optimize the sectioning of tobacco stems transformed through the agarose embeddi...

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Main Authors: Nisa Ihsani, Fenny Martha Dwivany, Sony Suhandono
Format: Article
Language:English
Published: Universitas Gadjah Mada, Yogyakarta 2023-09-01
Series:Indonesian Journal of Biotechnology
Subjects:
Online Access:https://jurnal.ugm.ac.id/ijbiotech/article/view/80853
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author Nisa Ihsani
Fenny Martha Dwivany
Sony Suhandono
author_facet Nisa Ihsani
Fenny Martha Dwivany
Sony Suhandono
author_sort Nisa Ihsani
collection DOAJ
description It is difficult to observe plant tissue sections transformed using the agroinfiltration method under a fluorescent microscope. This is due to the softness of the post‐transformation plant. This research was conducted to optimize the sectioning of tobacco stems transformed through the agarose embedding technique. Optimization was conducted at various agarose concentrations: 2%, 4%, and 6%, followed by five minutes of incubation at various temperatures: –80 °C, 4 °C, and 25 °C. The stems were then cut using a scalpel and examined under a fluorescence microscope. The results showed that the embedding method using 6% agarose was more effective at producing a tobacco stem section than 2% or 4% agarose. Meanwhile, incubation at 25 °C was better suited to the transformed tobacco stems than at 4 °C or –80 °C. Green Fluorescent Protein (GFP) could be determined under a fluorescent microscope when using the optimum method. Thus, the optimum method for creating sections of transformed tobacco stems by embedding was to use 6% agarose followed by incubation at 25 °C for 5 min. The optimum result can be applied to obtain a slight section of tobacco stem in order to observe a recombinant protein or other anatomical structures.
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spelling doaj.art-090d5e8d1c584e41a67c8bfffcbf96eb2023-10-26T07:03:38ZengUniversitas Gadjah Mada, YogyakartaIndonesian Journal of Biotechnology0853-86542089-22412023-09-0128315315710.22146/ijbiotech.8085334582A simple method of plant sectioning using the agarose embedding technique for screening intracellular green fluorescent proteinNisa Ihsani0Fenny Martha Dwivany1Sony Suhandono2Department of Biotechnology, Faculty of Science and Technology, Universitas Muhammadiyah Bandung. Soekarno‐Hatta Street No.752 Bandung, Cipadung Kidul, Panyileukan, Bandung City, West Java 40614, IndonesiaThe School of Life Sciences and Technology, Institut Teknologi Bandung. Ganesa Street No.10, Bandung 40132The School of Life Sciences and Technology, Institut Teknologi Bandung. Ganesa Street No.10, Bandung 40132It is difficult to observe plant tissue sections transformed using the agroinfiltration method under a fluorescent microscope. This is due to the softness of the post‐transformation plant. This research was conducted to optimize the sectioning of tobacco stems transformed through the agarose embedding technique. Optimization was conducted at various agarose concentrations: 2%, 4%, and 6%, followed by five minutes of incubation at various temperatures: –80 °C, 4 °C, and 25 °C. The stems were then cut using a scalpel and examined under a fluorescence microscope. The results showed that the embedding method using 6% agarose was more effective at producing a tobacco stem section than 2% or 4% agarose. Meanwhile, incubation at 25 °C was better suited to the transformed tobacco stems than at 4 °C or –80 °C. Green Fluorescent Protein (GFP) could be determined under a fluorescent microscope when using the optimum method. Thus, the optimum method for creating sections of transformed tobacco stems by embedding was to use 6% agarose followed by incubation at 25 °C for 5 min. The optimum result can be applied to obtain a slight section of tobacco stem in order to observe a recombinant protein or other anatomical structures.https://jurnal.ugm.ac.id/ijbiotech/article/view/80853agaroseembeddinggreen fluorescent proteinplant sectiontransformation
spellingShingle Nisa Ihsani
Fenny Martha Dwivany
Sony Suhandono
A simple method of plant sectioning using the agarose embedding technique for screening intracellular green fluorescent protein
Indonesian Journal of Biotechnology
agarose
embedding
green fluorescent protein
plant section
transformation
title A simple method of plant sectioning using the agarose embedding technique for screening intracellular green fluorescent protein
title_full A simple method of plant sectioning using the agarose embedding technique for screening intracellular green fluorescent protein
title_fullStr A simple method of plant sectioning using the agarose embedding technique for screening intracellular green fluorescent protein
title_full_unstemmed A simple method of plant sectioning using the agarose embedding technique for screening intracellular green fluorescent protein
title_short A simple method of plant sectioning using the agarose embedding technique for screening intracellular green fluorescent protein
title_sort simple method of plant sectioning using the agarose embedding technique for screening intracellular green fluorescent protein
topic agarose
embedding
green fluorescent protein
plant section
transformation
url https://jurnal.ugm.ac.id/ijbiotech/article/view/80853
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