Advanced Fluorescence Microscopy and Image Processing

By Prof.Gunjan Mehta | IIT Hyderabad

Learners enrolled: 1102 | Exam registration: 37

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 control

PREREQUISITES: 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.

Summary

Course Status :	Upcoming
Course Type :	Elective
Language for course content :	English
Duration :	12 weeks
Category :	Biotechnology Biosciences Bioengineering
Credit Points :	3
Level :	Undergraduate/Postgraduate
Start Date :	20 Jan 2025
End Date :	11 Apr 2025
Enrollment Ends :	27 Jan 2025
Exam Registration Ends :	14 Feb 2025
Exam Date :	26 Apr 2025 IST

Note: This exam date is subject to change based on seat availability. You can check final exam date on your hall ticket.

Page Visits

Course layout

Week 1:

Lecture 1: Introduction to fluorescence microscopy and its applications

Lecture 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, FLIM

Lecture 6: Quantifying protein dynamics using FRAP, FRET, and FLIM

Week 4:

Lecture 7: Illumination strategies (Epi, TIRF, HILO, confocal, light sheet, multi-photon)

Lecture 8: Confocal Microscopy

Week 5:

Lecture 9: Super-resolution microscopy (SIM, STED, STORM/PALM)
Lecture 10: Nanomaterials for cellular imaging

Week 6:

Lecture 11: Immunofluorescence (IF), Immunohistochemistry (IHC)

Lecture 12: Fluorescence in-situ Hybridization (FISH), RNAFISH

Week 7:

Lecture 13: Digital images and camera technologies for microscopy

Lecture 14: CCD, EMCCD, sCMOS camera

Week 8:

Lecture 15: ImageJ/FIJI-based image processing and data analysis (workshop, basic operations using ImageJ/FIJI, montage preparation)

Week 9:

Lecture 16: ImageJ/FIJI-based image processing and data analysis (workshop, image segmentation)

Week 10:

Lecture 17: ImageJ/FIJI-based image processing and data analysis (workshop, macros, intensity measurement, scale bar, time scale, colocalization)

Week 11:

Lecture 18: Single-molecule imaging and tracking

Lecture 19: Optical tweezers and traction force microscopy

Week 12:

Lecture 20: Spatial transcriptomics and proteomics (RNAscope, MERFISH, CODEX)

Lecture 21: High-content imaging
Lecture 22: Designing fluorescence microscopy experiments

Books and references

1) Microscope Image Processing by Qiang Wu, Fatima Merchant and Kenneth Castleman, Academic Press, ISBN: 012372578X

2) Fundamentals of Light Microscopy and Electronic Imaging by Douglas Murphy and Michael Davidson, Second Edition, Wiley-Blackwell publisher, ISBN: 047169214X.

3) Introduction to optical microscopy by Jerome Mertz, Cambridge University Press, 2nd Edition, doi: 10.1017/9781108552660, ISBN: 9781108552660

4) Fluorescence Microscopy: From Principles to Biological Applications by Ulrich Kubitscheck, 2nd edition, John Wiley & Sons publisher, eBook ISBN 9783527687725

e-resource: https://www.microscopyu.com/

e-resource: https://www.ibiology.org/online-biology-courses/microscopy-series/

e-resource: http://zeiss-campus.magnet.fsu.edu/index.html

e-resource: https://www.leica-microsystems.com/science-lab/topics/basics-in-microscopy/

e-resource: https://www.olympus-lifescience.com/en/microscope-resource/

e-resource: https://imagej.nih.gov/ij/docs/examples/index.html

Instructor bio

Prof.Gunjan Mehta

IIT Hyderabad

Dr. Mehta earned a B.Sc. (Biochemistry) from St. Xavier’s College, Ahmedabad, Gujarat, and M.Sc. (Microbiology) from the M.S. University of Baroda, Vadodara, Gujarat. During his Ph.D. at IIT Bombay (2009-2015), he used several advanced imaging technologies to study the mechanism of cell division. He worked as a post-doctoral research fellow (2015-2020) at the National Cancer Institute (NCI), National Institutes of Health (NIH), Bethesda, MD, USA. He worked for 5.5 years in the microscopy core facility of NCI and developed single-molecule imaging and tracking methods for quantifying protein dynamics in live yeast cells. He has published several high-impact research publications using advanced imaging technologies. He returned to India in 2020 and joined IIT Hyderabad as an Assistant Professor. Here, his research group develops single-molecule imaging tools to study the molecular mechanism of cell division/chromosome segregation and gene regulation. For more information on his current research projects, please visit www.mehtalab-iith.com.

Course certificate

The course is free to enroll and learn from. But if you want a certificate, you have to register and write the proctored exam conducted by us in person at any of the designated exam centres.

The exam is optional for a fee of Rs 1000/- (Rupees one thousand only).

Date and Time of Exams: April 26, 2025 Morning session 9am to 12 noon; Afternoon Session 2pm to 5pm.

Registration url: Announcements will be made when the registration form is open for registrations.

The online registration form has to be filled and the certification exam fee needs to be paid. More details will be made available when the exam registration form is published. If there are any changes, it will be mentioned then.

Please check the form for more details on the cities where the exams will be held, the conditions you agree to when you fill the form etc.

CRITERIA TO GET A CERTIFICATE

Average assignment score = 25% of average of best 8 assignments out of the total 12 assignments given in the course.

Exam score = 75% of the proctored certification exam score out of 100

Final score = Average assignment score + Exam score

Please note that assignments encompass all types (including quizzes, programming tasks, and essay submissions) available in the specific week.

YOU WILL BE ELIGIBLE FOR A CERTIFICATE ONLY IF AVERAGE ASSIGNMENT SCORE >=10/25 AND EXAM SCORE >= 30/75. If one of the 2 criteria is not met, you will not get the certificate even if the Final score >= 40/100.

Certificate will have your name, photograph and the score in the final exam with the breakup.It will have the logos of NPTEL and IIT Hyderabad .It will be e-verifiable at nptel.ac.in/noc.

Only the e-certificate will be made available. Hard copies will not be dispatched.

Once again, thanks for your interest in our online courses and certification. Happy learning.

- NPTEL team