Low-cost, scalable laser scanning module for real-time reflectance and fluorescence confocal microscopy.

We present a low-cost, high-speed, retrofitted laser scanning module for microscopy. The cage-mounted system, with various available fiber-coupled sources, offers a real-time imaging alternative to costly commercial systems with capabilities for conventional or confocal reflectance and fluorescence applications as well as advanced laser scanning microscopy implementations. Reflectance images of a resolution target and confocal images of fluorescent polystyrene beads are presented for system characterization. Confocal fluorescence image stacks of T84 epithelial cancer cells are presented to demonstrate application to biological studies. This laser scanning module is a flexible, scalable, high-speed alternative to commercial laser scanning systems suitable for applications requiring a simple imaging tool and for teaching laboratories.

[1]  Satoshi Kawata,et al.  Second-harmonic-generation microscope with a microlens array scanner. , 2002, Optics letters.

[2]  R Richards-Kortum,et al.  Fiber confocal reflectance microscope (FCRM) for in-vivo imaging. , 2001, Optics express.

[3]  S. Hell,et al.  Time-multiplexed multifocal multiphoton microscope. , 2001, Optics letters.

[4]  J. Fujimoto,et al.  Optical coherence microscopy in scattering media. , 1994, Optics letters.

[5]  Karsten König,et al.  Modern laser scanning microscopy in biology, biotechnology and medicine. , 2003, Annals of anatomy = Anatomischer Anzeiger : official organ of the Anatomische Gesellschaft.

[6]  W. Denk,et al.  Two-photon laser scanning fluorescence microscopy. , 1990, Science.

[7]  J. Ploem,et al.  Laser scanning fluorescence microscopy. , 1987, Applied optics.

[8]  K. Shi,et al.  Chromatic confocal microscopy using supercontinuum light. , 2004, Optics express.

[9]  R. Webb,et al.  Spectrally encoded confocal microscopy. , 1998, Optics letters.

[10]  S. Paddock.  Principles and practices of laser scanning confocal microscopy , 2000, Molecular biotechnology.

[11]  D Yelin,et al.  Laser scanning third-harmonic-generation microscopy in biology. , 1999, Optics express.

[12]  Retrofitted confocal laser scanner for a commercial inverted fluorescence microscope , 2001 .

[13]  P. Danielsson,et al.  Three-dimensional microscopy using a confocal laser scanning microscope. , 1985, Optics letters.

[14]  S. Paddock,et al.  Confocal laser scanning microscopy. , 1999, BioTechniques.

[15]  R Richards-Kortum,et al.  Fiber-optic confocal microscopy using a spatial light modulator. , 2000, Optics letters.

[16]  Satoshi Kawata,et al.  Confocal multipoint multiphoton excitation microscope with microlens and pinhole arrays , 2000 .

[17]  Yaron Silberberg,et al.  Multiphoton plasmon-resonance microscopy. , 2003, Optics express.

[18]  J G Fujimoto,et al.  High-resolution optical coherence microscopy for high-speed, in vivo cellular imaging. , 2003, Optics letters.

[19]  Kung-Bin Sung,et al.  Fiber optic confocal reflectance microscopy: a new real-time technique to view nuclear morphology in cervical squamous epithelium in vivo. , 2003, Optics express.

[20]  S W Hell,et al.  Two-photon near- and far-field fluorescence microscopy with continuous-wave excitation. , 1998, Optics letters.

[21]  H. Guyader,et al.  Confocal scanning optical microscopy and three‐dimensional imaging , 1992 .

[22]  Siavash Yazdanfar,et al.  Interferometric second harmonic generation microscopy. , 2004, Optics express.