Enhanced Arabidopsis Live Cell Imaging Obtained Through Affordable and Efficient Heating Devices

Original video: https://youtu.be/SOZBoNkmWBU

Temperature control is crucial for live cell imaging, particularly in studies involving plant responses to high ambient temperatures and thermal stress. This study presents the design, development, and testing of two cost-effective heating devices tailored for confocal microscopy applications: an aluminum heat plate and a wireless mini-heater. The aluminum heat plate, engineered to integrate seamlessly with the standard 160 mm × 110 mm microscope stage, supports temperatures up to 36°C, suitable for studies in the range of non-stressful warm temperatures (e.g., 25-27°C for Arabidopsis thaliana) and moderate heat stress (e.g., 30-36°C for A. thaliana). We also developed a wireless mini-heater that offers rapid, precise heating directly at the sample slide, with a temperature increase rate over 30 times faster than the heat plate. The wireless heater effectively maintained target temperatures up to 50°C, ideal for investigating severe heat stress and heat shock responses in plants. Both devices performed well in controlled studies, including the real-time analysis of heat shock protein accumulation and stress granule formation in A. thaliana. Our designs are effective and affordable, with total construction costs lower than $300. This accessibility makes them particularly valuable for small laboratories with limited funding. Future improvements could include enhanced heat uniformity, humidity control to mitigate evaporation, and more robust thermal management to minimize focus drift during extended imaging sessions. These modifications would further solidify the utility of our heating devices in live cell imaging, offering researchers reliable, budget-friendly tools for exploring plant thermal biology.

The video shows the time-lapse of YFP-AtHSP70-4 accumulation in Arabidopsis epidermis and hypocotyl treated with the heat plate at 36°C as well as the time-lapse of the transgenic Arabidopsis (YFP-eEF1Bβ) containing cytoplasmic foci in Arabidopsis hypocotyl treated with the wireless heater at 42°C. YFP-AtHSP70-4 refers to a genetically engineered construct used for live cell imaging experiments. Specifically, it is an Arabidopsis thaliana heat shock protein 70 (AtHSP70-4) that has been tagged at its N-terminus with a yellow fluorescent protein (YFP) and a 3xHA-tag. The expression of this tagged protein is controlled by its native promoter (AtHSP70-4pro::3 HA-YFP-AtHSP70-4), and it is abbreviated as YFP-HSP70 in the text.

Keywords: Time-Lapse, Arabidopsis, Cost-effective, Confocal microscopy, Live cell imaging, Heat stress, Heat shock protein, Microheater

Citation:

Bajracharya A, Timilsina S, Cao R, Jiang Q, Dickey BA, Wasti A, Xi J, Weingartner M, Baerson SR, Roman GW, Han Y and Qiu Y (2025) Developing affordable and efficient heating devices for enhanced live cell imaging in confocal microscopy. Front. Plant Sci. 15:1499831. doi: https://doi.org/10.3389/fpls.2024.1499831

Published on: 10 January 2025.

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