Developing affordable and efficient heating devices for enhanced live cell imaging in confocal microscopy

Authors: BAJRACHARYA, ABHISHESH - University Of Mississippi; TIMILSINA, SAMPADA - University Of Mississippi; CAO, RUOFAN - University Of Mississippi; JIANG, QINGRUI - University Of Mississippi; DICKEY, BERRY - University Of Mississippi; WASTI, ANUPA - University Of Mississippi; XI, JING - Orise Fellow; WEINGARTNER, MAGDALENA - University Of Hamburg; Baerson, Scott; QIU, YONGJIAN - University Of Mississippi; ROMAN, GREGG - University Of Mississippi; HAN, YIWEI - University Of Mississippi

Submitted to: Frontiers in Plant Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/20/2024
Publication Date: 1/10/2025
Citation: Bajracharya, A., Timilsina, S., Cao, R., Jiang, Q., Dickey, B.A., Wasti, A., Xi, J., Weingartner, M., Baerson, S.R., Qiu, Y., Roman, G.W., Han, Y. 2025. Developing affordable and efficient heating devices for enhanced live cell imaging in confocal microscopy. Frontiers in Plant Science. 15:1499831. https://doi.org/10.3389/fpls.2024.1499831.
DOI: https://doi.org/10.3389/fpls.2024.1499831

Interpretive Summary: Global warming represents unique challenges for biological investigations, particularly in the area of confocal microscopy-based live cell imaging requiring the precise application of increased temperatures to specific target cells. The present manuscript describes the development of heating devices designed to provide precise localized heat applications to live plant tissues during confocal microscopy. Test experiments were successfully performed using the wireless heating devices, and their effectiveness was demonstrated in an experiment examining heat shock protein accumulation and stress granule formation in Arabidopsis thaliana. These innovations will provide plant researchers with novel, cost-effective and reliable tools for exploring responses to elevated temperatures in live plant tissues in real time.

Technical Abstract: 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 (25-29°C) and moderate heat stress (30-36°C). 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 Arabidopsis 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.