BioTech 35: Immunofluorescence & Confocal Microscopy NIH Bldg. 162
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1 NIH Bldg. 162 Tuesday 8:40am 8:45am Welcome Jithesh Veetil (FAES) 8:45am 10:00am Fundamentals of Fluorescence Microscopy Scott Olenych (Zeiss) 10:00am 10:10am 10:10am 11:30am Intro to Immunofluorescence, and Its Practical Application Lynne Holtzclaw (NICHD) 11:30am 12:30pm 12:30pm 2:00pm Wet : Preparing a 3-color immunofluorescence slide for confocal microscopy, Part I Lynne Holtzclaw (NICHD) 2:00pm 3:30pm Basic Confocal Imaging Alma Arnold (Zeiss) 3:30pm 6:00pm Wet : Preparing a 3-color immunofluorescence slide for confocal microscopy, Part II Lynne Holtzclaw (NICHD)
2 Wednesday 9:00am 11:20am Confocal Basics I: Acquiring/Optimizing images: Effect of Pinhole, Gain, Laser Power/Attenuation, Objective Mag, NA, Scan Area, Pixel Size; Z-stacks See Mic Schedule 11:20am 12:20pm 12:20pm 1:20pm Confocal Basics II: Acquiring/Optimizing images: Effect of Pinhole, Gain, Laser Power/Attenuation, Objective Mag, NA, Scan Area, Pixel Size; Z-stacks See Mic Schedule 1:20pm 2:30pm Spinning Disk Confocal Demo See Mic Schedule 2:30pm 6:00pm Computer : Image Presentation/Analysis Primer Intro to Fiji; Viewing & Manipulating today s images Vincent Schram (NICHD)
3 Thursday 9:00am 9:45am Spinning Disk Confocal, Light Sheet Microscopy Valarie Barr (NCI) 9:45am 9:50am 9:50am 11:00am Super-Resolution down to 100nm Deconvolution; SIM (Structured Illumination); isim/airyscan Xufeng Wu (NHLBI) 11:00am 11:10am 11:10am 12:30pm CoLocalization Susan Garfield (NCI) 12:30pm 1:30pm 1:30pm 4:15pm Colocalization; Deconvolution; Spectral Unmixing See Mic Schedule 4:15pm 4:30pm 4:30pm 6:00pm Computer : Colocalization; Deconvolution; Spectral Unmixing Jeff Reece (NIDDK)
4 Friday 9:00am 10:00am FRET and BioSensors Tatiana Karpova (NCI) 10:00am 10:10am 10:10am 11:10am Live Cell Imaging: Fluorescent Proteins, FRAP, FLIP, Photo Activation Dani Cai (NICHD) 11:10am 11:20am Loose Ends; Feedback Jeff Reece / J. Veetil 11:20am 12:20pm 12:20pm 3:30pm FRET; FRAP See Mic Schedule 3:30pm 5:00pm Computer : FRET; FRAP Joe Brzostowski (NIAID) Valentin Jaumouillé (NHLBI)
5 MICROSCOPY LAB SCHEDULE For Wednesday-Friday microscopy labs, students will be divided into groups of no more than 4/group. Some effort will be made when assigning the grouping and the microscopes to accommodate the desires and interests of the students, as indicated on the Initial Questionnaires filled out and returned by Tuesday afternoon. The grouping assignments A-E will be indicated on a separate list passed out on Wednesday morning. The name of each student will have a letter beside it, e.g. Roger Tsien / A. Wednesday 4/19/2017 What Group Where Instructor 9-11:20am and 12:20-1:20pm: A 35/2C403: Zeiss LSM 880 Alma Arnold Confocal Basics: B 50/2315: Nikon A1R+ Jason Hill Acquiring/Optimizing images: Effect of Pinhole, C 5/329: Zeiss LSM 780 Edgar Vogt Gain, Laser Power/Attenuation, D 37/B114: Zeiss LSM 880 Christine Krieger Objective Mag, NA, Scan Area, Pixel Size Multi-color; Z-stacks E 49/3W16: Zeiss LSM 880 Nathanie Trisnadi A 8/B123: MetaMorph Jeff Reece 1:30-2:30pm: Spinning Disk Confocal Demo B 35A/GD922: Nikon Elements Jason Hill C 8/2A24: MetaMorph Simona Rosu D 37/2033B: MetaMorph Jim Paladino / Chris Anderson E 49/3B35: Zeiss ZEN Stephen Wincovitch Thursday 4/20/2017 What Group Where Instructor 1:30-4:15pm: Colocalization; Deconvolution; Spectral Unmixing A 35/2C403: Zeiss LSM 880 Alma Arnold B 37/2033B: Leica SP8 Owen Schwartz C 5/329: Zeiss LSM 780 Jeff Reece D 37/1035: Zeiss LSM 780 Javier Manzella-Lapeira E 35A/GD922: Zeiss LSM 780 Nathanie Trisnadi Friday 4/21/2017 What Group Where Instructor 12:20-3:30pm FRET; FRAP A 5/329: Zeiss LSM 780 Yehong Ye B 37/B114: Zeiss LSM710 NLO Joe Brzostowski C 4/B2-31c: Leica SP8 Val Jaumouillé D 49/3W16: Zeiss LSM 880 Dani Cai E 10/6C-208: Zeiss LSM 880 Javier Manzella-Lapeira
6 Special Thanks to the following Donors of Equipment NCI CCR Confocal Microscopy Core, Susan Garfield oratory of Genitourinary Cancer Pathogenesis Microscopy Core, Ross Lake oratory of Cellular and Molecular Biology Microscopy Core, Valarie Barr NHGRI Cytogenetic and Microscopy Core Facility, Stephen Wincovitch NHLBI Light Microscopy Core Building 10, Chris Combs Light Microscopy Core Building 50, Xufeng Wu NIAID Biological Imaging Facility, Owen Schwartz NICHD Microscopy and Imaging Core (MIC), Vincent Schram NIDDK Advanced Light Microscopy & Image Analysis Core (ALMIAC), Jeff Reece oratory of Cellular and Molecular Biology, John Hanover oratory of Biochemistry and Genetics, Kevin O'Connell NINDS Neurosciences Light Imaging Facility, Carolyn Smith
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