CVRI Microscopy Core: Summary of Capabilities
The CVRI Microscopy Core consists of 5 well-maintained microscopes. The Core is overseen by Drs. Peng Yao and Linda Callahan. The Microscopy Core works in conjunction with the CVRI Histology core and the microsurgery core for design and interpretation of microscopy-related experiments. The 5 microscopes within the core include a ZOE imaging system (ZOE), an upright epifluorescence microscope (BX51), an inverted epifluorescence microscope (IX 70), a timelapse/FRET microscope (capable of FRET, multipoint timelapse imaging, and TIRF), and a laser point scanning confocal microscope (FV1000 confocal). Dr. Linda Callahan trains all users on each of the microscopes and assists with experimental design and trouble-shooting of imaging experiments. Dr. Callahan also maintains the microscopes, working with vendors as needed to upgrade or maintain instruments. Below is a brief summary of the capabilities of each microscope.
- The ZOE imaging system allows for quick observation of cell cultures and slides. Best used for checking progress of culture expression or slide staining. Not for high resolution work, but excellent for quick checks of staining or expression progress. It is in room G.11300B.
- The BX51 microscope is capable of standard brightfield and epifluorescence microscopy, polarized light microscopy, phase contrast and differential interference microscopy. Full complement of objectives and a DP74 high resolution camera. Mercury lamp excitation and epifluorescence capabilities for blue to near red emitting fluorophores. Software includes measurement, post-processing, and ‘smart montage’ capabilities. Original meta-data with each image. It is in room G.11354.
- The IX70 microscope is capable of observation of cell culture and slides. Standard brightfield and epifluorescence. Mercury lamp excitation. It is in room G.11352.
- The timelapse/FRET microscope is capable of FRET, long-term multi-point timelapse, and TIRF imaging. Easily images live cell culture using glass bottom coverslip dishes or fixed tissue/fixed cultures. Fully automated. Inverted microscope with an enclosed temperature, humidity, and CO2 environmental control chamber. Multiple filter combination capabilities. Xenon lamp excitation and Hamamatsu Orca camera. It is in room G.11006.
- The FV1000 confocal microscope is a laser point scanning confocal microscope with high resolution imaging capabilities including 0.4 um slice capabilities using the 60x NA 1.42 objective. Applications include 2-4 channel 2D and 3D imaging, FRAP, FRET, timelapse, measurement, and post-processing capabilities. Original meta-data with each image. 6 laser lines (405, 440, 488, 515, 559, 635) enable imaging fluorophores emitting in the blue to far red spectrum. Floating table, in-room lighting adjustment capabilities, and secure one-way movement of images from the imaging computer to the CVRI confocal microscope server. Server is secured and designed to give PI full control over their data. It is in room G.11008.
Optimal imaging for the high-resolution microscopes is best when tissue samples or cultures are imaged through 0.17 mm coverslips for slides or 0.17 mm glass bottom dishes for cultures (#1.5 coverslips in catalogs). For more information, please contact Peng_Yao@urmc.rochester.edu and Linda_Callahan@urmc.rochester.edu.
Upper panel: Cellular localization of recombinant FAM210A in HEK293T cells. FAM210A-3xFLAG expression plasmid was transfected into HEK293T cells for 48 hrs. Anti-FLAG antibody was used for immunofluorescence. Mitotracker was used to label mitochondria.
Lower panel: Cellular localization of FAM210A in primary mouse adult cardiomyocytes detected by immunofluorescence using anti-FAM210A antibody. Wu J, Kadiam CVS, Jiang F, Hedaya O, Mohan A, Yang T, Welle K, Ghaemmaghami S, Tang WHW, Small E, Yan C, and Yao, P (2021). MicroRNA‐574 regulates FAM210A expression and influences pathological cardiac remodeling. EMBO Mol Med 13:e12710 https://doi.org/10.15252/emmm.202012710.
The above images have been published in EMBO Molecular Medicine paper on 12/28/2020 online as below. https://www.embopress.org/doi/full/10.15252/emmm.202012710.