Epi-Fluorescence Microscopy

 

The epi-fluorescence microscope experiments were designed in collaboration with Professor Mathias Lösche of the Institute of Experimental Physics, University of Leipzig, Germany.  The Langmuir film microscope is based on a design of Prof. Lösche's and has been previously described in several papers, for example, M. Lösche and H. Möhwald, "Fluorescence microscope to observe dynamical processes in monomolecular layers at the air/water interface", Rev. Sci. Instrum. 55 (1984) 1968-1972.   The microscope is based on a commercial Zeiss epi-fluorescence microscope arranged on top of a Langmuir trough.  A diagram of the design is shown below.

 

fluorescence diagram 2.eps (7089659 bytes)

 

Examples of some epi-fluorescent monolayer images

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Epi-fluorescence micrograph of phosphocholine (PC) and phosphoglycerol (PG) lipids at the air-water interface.  Surface pressure is approximately 13 mN/m.

 

 

 

 

 

fig3d_mo.JPG (29659 bytes)

 

Epi-fluorescence micrograph of phosphocholine (PC) and phosphoglycerol (PG) lipids at the air-water interface  with 1 wt% of the hydrophobic surfactant proteins SP-B/C added to the monolayer.  Surface pressure is approximately 15 mN/m.

 

 

 

 

 

Publications

We have published several papers that use fluorescence imaging of biophysical monolayers to investigate morphology and mechanisms in biophysical interfaces.  These include:

 

P. Krüger, M. Schalke, Z. Wang, R. Notter, M. Lösche and R.A. Dluhy, "Effect of Hydrophobic Surfactant Proteins SP-B and SP-C on Binary Phospholipid Monolayers. I. Fluorescence and Dark Field Microscopy", Biophysical Journal 1999, 77 903-914.  [pdf]

P. Krüger, J.E. Baatz, R.A. Dluhy and M. Lösche, “Effect of the Hydrophobic Surfactant Peptide SP-C on Binary Phospholipid Monolayers.  Molecular Machinery at the Air-Water Interface”, Biochemical Journal 2002, 99, 209-228.  [pdf]

N. Biswas, S. Shanmukh, A. Waring, F. Walther, Z. Wang, Y. Chang, R. Notter and R.A. Dluhy, “Structure And Morphology Of Phospholipid-Peptide Monolayers Containing Monomeric SP-B1-25.  I.  Phases and Morphology by Epi-Fluorescence Microscopy”, Biophysical Chemistry 2005, 113, 223-232.  [pdf]