Femtosecond Pulsed Laser as a Microscalpel for Microdissection and Isolation of Specific Sections from Biological Samples

The femtosecond laser is a candidate tool for dissecting and obtaining specific tissues or cells from biological samples. In this paper, we report on the dissection of flammable (heat-sensitive) plant tissues (Eucommia ulmoides stem) and that of a transparent zebrafish embryo placed in water. During laser dissection in the plant tissues, we observed that a low repetition rate of the irradiations is crucial for reducing the risk of thermal damage. During laser dissection of the zebrafish embryo, the laser-induced bubble formation at the tissue surface disturbed the laser-induced dissection. We concluded that the "interior laser dissection method", in which the dissection is performed by moving the laser spot inside a biological sample, is an efficient way to dissect a sample that is placed in water. In view of our findings, the advantages of fs laser dissection are discussed on the basis of the physical characteristics of fs laser machining.

[1]  Yaroslava G Yingling,et al.  Computer simulations of laser ablation of molecular substrates. , 2003, Chemical reviews.

[2]  D E Ingber,et al.  Pulse energy dependence of subcellular dissection by femtosecond laser pulses. , 2005, Optics express.

[3]  Masaaki Sakakura,et al.  Initial temporal and spatial changes of the refractive index induced by focused femtosecond pulsed laser irradiation inside a glass , 2005 .

[4]  E. Mazur,et al.  Bulk heating of transparent materials using a high-repetition-rate femtosecond laser , 2003 .

[5]  K. Miura,et al.  Writing waveguides in glass with a femtosecond laser. , 1996, Optics letters.

[6]  A. Vogel,et al.  Mechanisms of pulsed laser ablation of biological tissues. , 2003, Chemical reviews.

[7]  L. Liotta,et al.  Laser Capture Microdissection , 1996, Science.

[8]  R. Storb,et al.  Selective Mitochondrial Damage by a Ruby Laser Microbeam: An Electron Microscopic Study , 1965, Science.

[9]  Kazuyoshi Itoh,et al.  Femtosecond laser disruption of subcellular organelles in a living cell. , 2004, Optics express.

[10]  Karsten König,et al.  Laser tweezers and multiphoton microscopes in life sciences , 2000, Histochemistry and Cell Biology.

[11]  K. Fukui,et al.  Microdissection of plant chromosomes by argon-ion laser beam , 1992, Theoretical and Applied Genetics.

[12]  Iris Riemann,et al.  Intratissue surgery with 80 MHz nanojoule femtosecond laser pulses in the near infrared. , 2002, Optics express.

[13]  Barry Luther-Davies,et al.  Ultrafast ablation with high-pulse-rate lasers. Part I: Theoretical considerations , 1999 .

[14]  D. Kleinfeld,et al.  Targeted insult to subsurface cortical blood vessels using ultrashort laser pulses: three models of stroke , 2006, Nature Methods.

[15]  N. Inada,et al.  Novel tissue preparation method and cell-specific marker for laser microdissection of Arabidopsis mature leaf , 2005, Planta.

[16]  I Itzkan,et al.  The thermoelastic basis of short pulsed laser ablation of biological tissue. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[17]  J. P. Callan,et al.  Three-dimensional optical storage inside transparent materials. , 1996, Optics letters.

[18]  D. Kleinfeld,et al.  All-Optical Histology Using Ultrashort Laser Pulses , 2003, Neuron.

[19]  Günther Paltauf,et al.  Photomechanical processes and effects in ablation. , 2003, Chemical reviews.

[20]  Vikram Kohli,et al.  Cell nanosurgery using ultrashort (femtosecond) laser pulses: Applications to membrane surgery and cell isolation , 2005, Lasers in surgery and medicine.

[21]  A. Vogel,et al.  Mechanisms of femtosecond laser nanosurgery of cells and tissues , 2005 .

[22]  R. Krueger,et al.  First clinical results with the femtosecond neodynium-glass laser in refractive surgery. , 2003, Journal of refractive surgery.

[23]  Fumiyo Yoshino,et al.  Heat accumulation effects in femtosecond laser-written waveguides with variable repetition rate. , 2005, Optics express.

[24]  K König,et al.  Nanodissection of human chromosomes with near-infrared femtosecond laser pulses. , 2001, Optics letters.

[25]  Ernst H. K. Stelzer,et al.  Ultraviolet diffraction limited nanosurgery of live biological tissues , 2004 .