Final Project -- Nathan S Lachenmyer
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Revision as of 01:13, 13 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.
Goal
- Minimum: Create a z-stack of images of a 3D sample
- Potential Improvements:
- 3D Reconstruction in Software
- Laser Scanning for CLSM
Proposed Design
- Aperture to block out non-convergent rays
Challenges
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
- Determine Ca$ ^{2+} $ concentration in a biological system
- Potential Improvements:
- Lifetime vs. Intensity Measurements
- FRET Measurements
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
