Final Project -- Nathan S Lachenmyer
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Revision as of 16:54, 15 March 2012 by Nathan S Lachenmyer (Talk | contribs)
I have two project proposals that I am deciding between:
Contents
Confocal Microscopy
Literature
- Imaging in Neuroscience and Development: A Laboratory Manual / editors, Rafael Yuste and Arthur Konnerth
- Confocal microscopy for biologists / Alan R. Hibbs.
- Handbook of biological confocal microscopy / editor, James B. Pawley.
Goals and Checkpoints
- 2 Weeks: Brightfield Confocal Microscopy
- 2 Weeks: Software Image Capture, PSF Characterized, Z-Stack Images
- 3 Weeks: Laser Scanning Features
Proposed Design
Challenges
- Control software
- Getting good resolution
Ca$ ^{2+} $ Imaging
Literature
- Imaging in Neuroscience and Development: A Laboratory Manual / editors, Rafael Yuste and Arthur Konnerth
- Handbook of biological confocal microscopy / editor, James B. Pawley.
- Detection and Measurement of Free Ca2+ in Cells / editor, Ashley Campbell
- Calcium Measurement Methods / editors, Alexei Verkhratsky and Ole Peterson
Goals and Checkpoints
- 1 Weeks: Fluorescence Microscopy Setup (with UV)
- 2 Weeks: Software Image Capture, PSF Characterized
- 2 Weeks: Switching between UV Sources implemented, Image Analysis Done
- 2 Weeks: [Ca] samples characterized
Proposed Design
- Fluorescence Microscope
- 422 nm (brightfield, optical)
- 340, 380 nm (fluorescence; two wavelengths provides ratiometric determination of [Ca])
- Motorized Mirror or Filter (Servo) to switch wavelengths
- Sync motor signal with camera signal (take pictures)
- Post-Processing Analysis to determine [Ca](t)
Challenges
- Syncing up motor with camera for fluorescence imaging
- Acquiring UV Optics
- Biggest Risk: Biology doesn't work out
