Difference between revisions of "Final Project -- Nathan S Lachenmyer"

From Course Wiki
Jump to: navigation, search
(Ca^{2+} Imaging)
Line 32: Line 32:
  
 
==Proposed Design==
 
==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=
 
==Challenges=
 
* Syncing up motor with camera for fluorescence imaging
 
* Syncing up motor with camera for fluorescence imaging

Revision as of 01:11, 13 March 2012

I have two project proposals that I am deciding between:

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

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