18350427161Table: Instruments and their usage in the FACS Laboratory (2020)Instrument typesFACS cell analyzerFACS cell sorterMass cytometer(1)(4)Figure: Leica SP8 confocal microscope (1), Leica SP8 multiphoton micro-scope (2), Nikon N-SIM/N-STORM super-resolution microscope (4), GE Healthcare DeltaVision Elite system (5), Keyence BZ-X700 microscope (6), and Hitachi Regulus8420 FE-SEM (7)with access to the most advanced equipment and technolo-gies. Central Facilities consist of four sections; the FACS Labora-tory managed by Dr. Takashi Saito, the Microscope Laboratory cytometry and cell sorting techniques that are essential for nearly all experiments in immunology, genome research and dis-ease studies. The Laboratory supports both population and sin-gle-cell analysis and has upgraded all FACS Aria instruments, in-cluding two Aria Fusions, for multi-color analyses. In addition to FACS instruments, the lab possesses a mass spectrometry-based cytometer, HELIOS, which has the potential to analyze more than 40 markers simultaneously with metal-labeled antibodies.In 2020, even during the difficult and restricted period due to COVID-19, 510 analytical and 925 sorting experiments with FACS and 5 analyses with HELIOS were performed in the Laboratory.In the FACS laboratory, a specialized staff member offers vari-ous services for users of the equipment (cell analyzers and cell sorters): (1) Technical support and training: In 2020, the facility offered eight technical courses (four for cell sorting and four for cell analysis). The courses were held at three different levels, Cali-bur basic (1), Canto II (2) and Aria basic (4). An Aria course was also held in English. A total of 42 researchers participated in these tissue imaging and coordinates technical support. There are 6 fluorescence microscopes and 1 scanning electron microscope available to researchers at IMS.(1) Inverted Leica SP8 system equipped with hybrid detectors and the LIGHTNING super-resolution image extraction module.(2) Inverted Leica SP8 system with two femtosecond Ti:Sa lasers for multiphoton excitation. This system is equipped with two types of scanners (resonant and galvano) and hybrid detectors. One of the two Ti:Sa lasers is connected to an optical paramet-ric oscillator (OPO), which enables two-photon imaging by long wavelength excitation.(3) Inverted Leica SP5 system with hybrid detectors.(4) Inverted Nikon N-SIM/N-STORM super-resolution micro-scope for dual color imaging.(5) GE Healthcare DeltaVision Elite system.(6) Keyence BZ-X700 all-in-one fluorescence microscope.(7) Hitachi field emission scanning electron microscope (FE-SEM) Regulus8240.managed by Dr. Takaharu Okada, the Genomics Laboratory man-aged by Drs. Yukihide Momozawa and Jun Seita, and the Animal Facility managed by Dr. Haruhiko Koseki.courses in 2020. (2) Cell sorting operation service: The Laboratory provides a cell sorting operation service, in which researchers can ask an experienced operator to conduct the sorting experiment. In 2020, we provided 120 such services. Advanced cell sorting techniques, such as single cell sorting, have also been performed. (3) Management/ maintenance of FACS instruments: FACS ma-chines are available for registered users around the clock and reservations are accepted up to one month in advance through an internal website. In addition to the in-house FACS Laboratory staff, engineers from Becton Dickinson visit once a week to pro-vide maintenance and technical support.ModelmachinesCaliburCanto IIAria IIIu/III/FusionCyTOF2(2)(5)# of # of users55059255# of training sessions(6)(7)FACS LaboratoryThe FACS Laboratory provides a range of support for flow Microscope LaboratoryThe Microscope Laboratory provides equipment for cell and Central FacilitiesCentral Facilities in IMS provide all researchers in the Center
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