Laser-scanning coherent anti-stokes Raman scattering microscopy and applications to cell biology

被引:343
作者
Cheng, JX
Jia, YK
Zheng, GF
Xie, XS
机构
[1] Harvard Univ, Dept Chem & Biol Chem, Cambridge, MA 02138 USA
[2] Olympus Amer Inc, SEG, Melville, NY 11747 USA
关键词
D O I
10.1016/S0006-3495(02)75186-2
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
Laser-scanning coherent anti-Stokes Raman scattering (CARS) microscopy with fast data acquisition and high sensitivity has been developed for vibrational imaging of live cells. High three-dimensional (3D) resolution is achieved with two collinearly overlapped near infrared picosecond beams and a water objective with a high numerical aperture. Forward-detected CARS (F-CARS) and epi-detected CARS (E-CARS) images are recorded simultaneously. F-CARS is used for visualizing features comparable to or larger than the excitation wavelength, while E-CARS allows detection of smaller features with a high contrast. F-CARS and E-CARS images of live and unstained cells reveal details invisible in differential interference-contrast images. High-speed vibrational imaging of unstained cells undergoing mitosis and apoptosis has been carried out. For live NIH 3T3 cells in metaphase, 3D distribution of chromosomes is mapped at the frequency of the DNA backbone Raman band, while the vesicles surrounding the nucleus is imaged by E-CARS at the frequency of the C-H stretching Raman band. Apoptosis in NIH 3T3 cells is monitored using the CARS signal from aliphatic C-H stretching vibration.
引用
收藏
页码:502 / 509
页数:8
相关论文
共 21 条
[1]   Nonlinear scanning laser microscopy by third harmonic generation [J].
Barad, Y ;
Eisenberg, H ;
Horowitz, M ;
Silberberg, Y .
APPLIED PHYSICS LETTERS, 1997, 70 (08) :922-924
[2]   EFFECTS OF FOCUSING ON 3RD-ORDER NONLINEAR PROCESSES IN ISOTROPIC MEDIA [J].
BJORKLUND, GC .
IEEE JOURNAL OF QUANTUM ELECTRONICS, 1975, QE11 (06) :287-296
[3]   CHARACTERIZATION OF APOPTOTIC PHENOMENA INDUCED BY TREATMENT WITH L-ASPARAGINASE IN NIH3T3 CELLS [J].
BUSSOLATI, O ;
BELLETTI, S ;
UGGERI, J ;
GATTI, R ;
ORLANDINI, G ;
DALLASTA, V ;
GAZZOLA, GC .
EXPERIMENTAL CELL RESEARCH, 1995, 220 (02) :283-291
[4]   High-resolution nonlinear optical imaging of live cells by second harmonic generation [J].
Campagnola, PJ ;
Wei, MD ;
Lewis, A ;
Loew, LM .
BIOPHYSICAL JOURNAL, 1999, 77 (06) :3341-3349
[5]   Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues [J].
Campagnola, PJ ;
Millard, AC ;
Terasaki, M ;
Hoppe, PE ;
Malone, CJ ;
Mohler, WA .
BIOPHYSICAL JOURNAL, 2002, 82 (01) :493-508
[6]   Theoretical and experimental characterization of coherent anti-Stokes Raman scattering microscopy [J].
Cheng, JX ;
Volkmer, A ;
Xie, XS .
JOURNAL OF THE OPTICAL SOCIETY OF AMERICA B-OPTICAL PHYSICS, 2002, 19 (06) :1363-1375
[7]   Polarization coherent anti-Stokes Raman scattering microscopy [J].
Cheng, JX ;
Book, LD ;
Xie, XS .
OPTICS LETTERS, 2001, 26 (17) :1341-1343
[8]   An epi-detected coherent anti-stokes raman scattering (E-CARS) microscope with high spectral resolution and high sensitivity [J].
Cheng, JX ;
Volkmer, A ;
Book, LD ;
Xie, XS .
JOURNAL OF PHYSICAL CHEMISTRY B, 2001, 105 (07) :1277-1280
[9]  
Cooper GM., 2000, The Development and Causes of Cancer, V2nd
[10]   2-PHOTON LASER SCANNING FLUORESCENCE MICROSCOPY [J].
DENK, W ;
STRICKLER, JH ;
WEBB, WW .
SCIENCE, 1990, 248 (4951) :73-76