Progress toward imaging biological samples with NSOM

Advancements in near-field scanning optical microscopy (NSOM) tip design as well as an interferometric feedback mechanism are presented for the common goal of imaging living biological samples under physiological conditions. The ability of a cantilevered tip to track the subtle topography changes of a fragile lipid film in an aqueous environment is demonstrated. In order to further the imaging capabilities, the probes have been chemically etched to reduce the spring constants of the tips, thereby lowering the forces imparted on the sample. An optical feedback mechanism used as an alternative to the conventional force feedback is also described. Utilizing this optical feedback mechanism, images have been obtained of fixed cells underwater. Finally, progress towards modifying the NSOM tip for chemical sensor applications is discussed in the context of eventually measuring ion fluxes through single protein channels. Together these advancements demonstrate the potential of NSOM for studying live cells.

[1]  M Ohtsu,et al.  Reproducible fabrication of a fiber probe with a nanometric protrusion for near-field optics. , 1997, Applied optics.

[2]  Robert J. Chichester,et al.  Single Molecules Observed by Near-Field Scanning Optical Microscopy , 1993, Science.

[3]  N. F. van Hulst,et al.  Biological applications of near-field optical microscopy , 1996 .

[4]  Dieter W. Pohl,et al.  Near‐field optical scanning microscopy in reflection , 1988 .

[5]  R. Kopelman,et al.  Submicrometer intracellular chemical optical fiber sensors. , 1992, Science.

[6]  Claude Philipona,et al.  Fiber tips for scanning near-field optical microscopy fabricated by normal and reverse etching , 1998 .

[7]  Ci-Ling Pan,et al.  A simple chemical etching technique for reproducible fabrication of robust scanning near-field fiber probes , 1998 .

[8]  J. Bishop,et al.  Directed establishment of rat brain cell lines with the phenotypic characteristics of type 1 astrocytes. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[9]  Frederick Lanni,et al.  Near‐field fluorescence imaging of cytoskeletal actin , 1993 .

[10]  S. Buratto,et al.  A Reflection Near-Field Scanning Optical Microscope Technique for Subwavelength Resolution Imaging of Thin Organic Films , 1997 .

[11]  B. Streetman,et al.  A design of reflection scanning near-field optical microscope and its application to AlGaAs/GaAs heterostructures , 1996 .

[12]  D. Chemla,et al.  Near-field fluorescence microscopy of cells. , 1998, Ultramicroscopy.

[13]  H. Mcconnell,et al.  Supported phospholipid bilayers. , 1985, Biophysical journal.

[14]  A J Malkin,et al.  Atomic force microscopy studies of living cells: visualization of motility, division, aggregation, transformation, and apoptosis. , 1997, Journal of structural biology.

[15]  Levi A. Gheber,et al.  Domains in cell plasma membranes investigated by near-field scanning optical microscopy. , 1998, Biophysical journal.

[16]  R. Kopelman,et al.  Analytical properties and sensor size effects of a micrometer-sized optical fiber glucose biosensor. , 1996, Analytical chemistry.

[17]  Hirosato Monobe,et al.  Multi-functional SNOM/AFM probe with accurately controlled low spring constant , 1998 .

[18]  Motoichi Ohtsu,et al.  Fabrication of a triple tapered probe for near-field optical spectroscopy in UV region based on selective etching of a multistep index fiber , 1998 .

[19]  R. Weis,et al.  Periodic structures in lipid monolayer phase transitions. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[20]  Robert C. Dunn,et al.  HIGH RESOLUTION FLUORESCENCE IMAGING WITH CANTILEVERED NEAR-FIELD FIBER OPTIC PROBES , 1996 .

[21]  M. Isaacson,et al.  Probe-sample interactions in reflection near-field scanning optical microscopy. , 1995, Applied optics.

[22]  A. H. Klahn,et al.  References and Notes , 2022 .

[23]  J M Simard,et al.  The patch clamp technique. , 1995, Neurosurgery.

[24]  R. Dunn,et al.  Single molecule detection and underwater fluorescence imaging with cantilevered near-field fiber optic probes , 1998 .

[25]  E. Sackmann,et al.  A Fluorescence Microscopic Study Concerning the Phase Diagram of Phospholipids , 1983 .

[26]  S. Schaus,et al.  Cell viability and probe-cell membrane interactions of XR1 glial cells imaged by atomic force microscopy. , 1997, Biophysical journal.

[27]  R. Guckenberger,et al.  Scanning near-field fluorescence microscopy of thin organic films at the water/air interface , 1998 .