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

被引:0
|
作者
Bajracharya, Abhishesh [1 ]
Timilsina, Sampada [1 ]
Cao, Ruofan [2 ]
Jiang, Qingrui [3 ]
Dickey, Berry A. [1 ]
Wasti, Anupa [1 ]
Xi, Jing [4 ]
Weingartner, Magdalena [5 ]
Baerson, Scott R. [4 ]
Roman, Gregg W. [2 ]
Han, Yiwei [3 ]
Qiu, Yongjian [1 ]
机构
[1] Univ Mississippi, Dept Biol, University, MS 38677 USA
[2] Univ Mississippi, Sch Pharm, Dept Biomol Sci, University, MS 38677 USA
[3] Univ Mississippi, Dept Mech Engn, University, MS 38677 USA
[4] US Dept Agr, Agr Res Serv, Nat Prod Utilizat Res Unit, University, MS USA
[5] Univ Hamburg, Inst Plant Sci & Microbiol, Hamburg, Germany
来源
基金
美国国家科学基金会; 美国国家卫生研究院;
关键词
cost-effective; low-maintenance; confocal microscopy; live cell imaging; heat stress; heat shock protein; stress granules (SG); microheater; TEMPERATURE CONTROL; RESPONSES; STRESS;
D O I
10.3389/fpls.2024.1499831
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
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 x 110 mm microscope stage, supports temperatures up to 36 degrees C, suitable for studies in the range of non-stressful warm temperatures (e.g., 25-27 degrees C for Arabidopsis thaliana) and moderate heat stress (e.g., 30-36 degrees 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 degrees 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.
引用
收藏
页数:16
相关论文
共 50 条
  • [1] Swept Field Laser Confocal Microscopy for Enhanced Spatial and Temporal Resolution in Live-Cell Imaging
    Castellano-Munoz, Manuel
    Peng, Anthony Wei
    Salles, Felipe T.
    Ricci, Anthony J.
    MICROSCOPY AND MICROANALYSIS, 2012, 18 (04) : 753 - 760
  • [2] Live Confocal Imaging of Developing Arabidopsis Flowers
    Prunet, Nathanael
    JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, 2017, (122):
  • [3] Label-Free Live-Cell Imaging with Confocal Raman Microscopy
    Klein, Katharina
    Gigler, Alexander M.
    Aschenbrenne, Thomas
    Monetti, Roberto
    Bunk, Wolfram
    Jamitzky, Ferdinand
    Morfill, Gregor
    Stark, Robert W.
    Schlegel, Juergen
    BIOPHYSICAL JOURNAL, 2012, 102 (02) : 360 - 368
  • [4] Combined FLIM, Confocal Microscopy, and STED Nanoscopy for Live-Cell Imaging
    Benard, Magalie
    Chamot, Christophe
    Schapman, Damien
    Lebon, Alexis
    Galas, Ludovic
    BIO-PROTOCOL, 2025, 15 (04):
  • [5] Time-lapse live-cell imaging of pyroptosis by confocal microscopy
    Jiang, Shuai
    Qin, Kunpeng
    Sun, Li
    STAR PROTOCOLS, 2023, 4 (04):
  • [6] Photonic crystal enhanced microscopy for imaging of live cell adhesion
    Chen, Weili
    Long, Kenneth D.
    Lu, Meng
    Chaudhery, Vikram
    Yu, Hojeong
    Choi, Ji Sun
    Polans, James
    Zhuo, Yue
    Harley, Brendan A. C.
    Cunningham, Brian T.
    ANALYST, 2013, 138 (20) : 5886 - 5894
  • [7] Live Cell Visualization of Golgi Membrane Dynamics by Super-resolution Confocal Live Imaging Microscopy
    Kurokawa, Kazuo
    Ishii, Midori
    Suda, Yasuyuki
    Ichihara, Akira
    Nakano, Akihiko
    METHODS FOR ANALYSIS OF GOLGI COMPLEX FUNCTION, 2013, 118 : 235 - 242
  • [8] Live cell imaging using confocal microscopy induces intracellular calcium transients and cell death
    Knight, MM
    Roberts, SR
    Lee, DA
    Bader, DL
    AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY, 2003, 284 (04): : C1083 - C1089
  • [9] Enhanced live cell imaging via photonic crystal enhanced fluorescence microscopy
    Chen, Weili
    Long, Kenneth D.
    Yu, Hojeong
    Tan, Yafang
    Choi, Ji Sun
    Harley, Brendan A.
    Cunningham, Brian T.
    ANALYST, 2014, 139 (22) : 5954 - 5963
  • [10] Physical chemistry in a single live cell: confocal microscopy
    Amin, Md. Asif
    Nandi, Somen
    Mondal, Prasenjit
    Mahata, Tanushree
    Ghosh, Surajit
    Bhattacharyya, Kankan
    PHYSICAL CHEMISTRY CHEMICAL PHYSICS, 2017, 19 (20) : 12620 - 12627