ABOUT THE COURSE:Fluorescence microscopy-based quantitative imaging plays a crucial role in life science research as it makes the cellular processes and structures visible. Recent advancements have made it possible to visualize cells/tissues/organoids in 6 dimensions (x, y, z, time, multi-color, multi-point), visualize cellular structures with super-resolution, quantify the dynamics of DNA and proteins at the single-molecule level, and measure the cellular forces at the piconewton level. This course offers the basics of fluorescence microscopy and its applications in life science research, digital images and camera technologies used in microscopy, and advanced imaging technologies. The practical component of this course will familiarize students with a free web tool, ImageJ/FIJI, for image processing and quantifying biological information from images.INTENDED AUDIENCE :B.Sc. / B.Tech / B.E. / M.Sc ./ M.Tech / M.E. / M.Pharm. / Ph.D. in life sciences / Biotechnology, post-docs, MBBS, any person employed in industry / academic research / quality controlPREREQUISITES: A Bachelor's degree in any area of life sciences (zoology, botany, biochemistry, biotechnology, microbiology, medical biotechnology, genetics, etc.) is preferred. However, final year B.Sc./B.Tech/B.E. students in life sciences/Biotechnology can also enroll.INDUSTRY SUPPORT: Reliance Life Sciences, Dr. Reddy’s Lab, Novartis, Bharat Biotech, Biocon, Syngene, Serum Institute of India, and many more companies in the field of life sciences/biotech.
Overview
Syllabus
Week 1:Lecture 1: Introduction to fluorescence microscopy and its applicationsLecture 2: Introduction to Microscope Optics
Week 2:Lecture 3: Design of a Fluorescence Microscope
Lecture 4: Fluorescent proteins, organic dyes, and protein labeling strategies Week 3:Lecture 5: 6D imaging, live cell imaging, time-lapse imaging, FRAP, FRET, FLIMLecture 6: Illumination strategies (Epi, TIRF, HILO, light sheet, multi-photon)
Week 4:Lecture 7: Confocal MicroscopyLecture 8: Super-resolution microscopy (SIM, STED, STORM/PALM)
Week 5:Live demonstration of a fluorescence microscope, 6D imaging of live cells
Week 6:Lecture 9: Immunofluorescence (IF), Immunohistochemistry (IHC)Lecture 10: Fluorescence in-situ Hybridization (FISH), RNAFISH
Week 7:Lecture 11: Digital images and camera technologies for microscopyLecture 12: CCD, EMCCD, sCMOS camera
Week 8:Lecture 13: ImageJ/FIJI-based image processing and data analysis
(workshop, basic operations of ImageJ/FIJI)
Week 9:Lecture 14: ImageJ/FIJI-based image processing and data analysis
(workshop, image segmentation)
Week 10:Lecture 15: ImageJ/FIJI-based image processing and data analysis (workshop, macros, intensity measurement, scale bar, time scale, montage preparation, colocalization)
Week 11:Lecture 16: Single-molecule imaging and trackingLecture 17: Optical tweezers and traction force microscopy
Week 12:Lecture 18: Spatial Transcriptomics and Proteomics (RNAscope, MERFISH, CODEX)Lecture 19: High-contentimaging
Week 2:Lecture 3: Design of a Fluorescence Microscope
Lecture 4: Fluorescent proteins, organic dyes, and protein labeling strategies Week 3:Lecture 5: 6D imaging, live cell imaging, time-lapse imaging, FRAP, FRET, FLIMLecture 6: Illumination strategies (Epi, TIRF, HILO, light sheet, multi-photon)
Week 4:Lecture 7: Confocal MicroscopyLecture 8: Super-resolution microscopy (SIM, STED, STORM/PALM)
Week 5:Live demonstration of a fluorescence microscope, 6D imaging of live cells
Week 6:Lecture 9: Immunofluorescence (IF), Immunohistochemistry (IHC)Lecture 10: Fluorescence in-situ Hybridization (FISH), RNAFISH
Week 7:Lecture 11: Digital images and camera technologies for microscopyLecture 12: CCD, EMCCD, sCMOS camera
Week 8:Lecture 13: ImageJ/FIJI-based image processing and data analysis
(workshop, basic operations of ImageJ/FIJI)
Week 9:Lecture 14: ImageJ/FIJI-based image processing and data analysis
(workshop, image segmentation)
Week 10:Lecture 15: ImageJ/FIJI-based image processing and data analysis (workshop, macros, intensity measurement, scale bar, time scale, montage preparation, colocalization)
Week 11:Lecture 16: Single-molecule imaging and trackingLecture 17: Optical tweezers and traction force microscopy
Week 12:Lecture 18: Spatial Transcriptomics and Proteomics (RNAscope, MERFISH, CODEX)Lecture 19: High-contentimaging
Taught by
Prof.Gunjan Mehta
