Intravital imaging of green fluorescent protein using two-photon laser-scanning microscopy

被引:89
作者
Potter, SM
Wang, CM
Garrity, PA
Fraser, SE
机构
[1] CALTECH,BECKMAN INST 13974,DIV BIOL,PASADENA,CA 91125
[2] UNIV CALIF LOS ANGELES,HOWARD HUGHES MED INST,MED RES LAB,LOS ANGELES,CA 90095
关键词
confocal; GFP; 2-photon; Drosophila R-cell; rat; hippocampal neuron;
D O I
10.1016/0378-1119(95)00681-8
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
Imaging a fluorophore in a living tissue presents several unique problems, The fluorescence from the labeled cell(s) may be weak, the labeled cells may be buried deep within tissue and the presence of a fluorophore may render the cells photo-sensitive. Two-photon laser-scanning microscopy (TPLSM) offers several advantages in meeting these challenges. We show that TPLSM provides greater sensitivity, better resolution and less photo-bleaching, as compared to confocal laser-scanning microscopy, The dramatically reduced photo-bleaching makes it possible to image cells continuously for long periods of time. Therefore, TPLSM allows a safer and higher-resolution means of imaging living cells labeled with a variety of fluorophores, including green fluorescent protein.
引用
收藏
页码:25 / 31
页数:7
相关论文
共 23 条
[1]  
Ausubel FM., 1994, Curr. Protoc. Mol. Biol
[2]   GREEN FLUORESCENT PROTEIN AS A MARKER FOR GENE-EXPRESSION [J].
CHALFIE, M ;
TU, Y ;
EUSKIRCHEN, G ;
WARD, WW ;
PRASHER, DC .
SCIENCE, 1994, 263 (5148) :802-805
[3]   MESSY FIBER GROWTH AND SYNAPTOGENESIS IN RAT HIPPOCAMPAL SLICES IN-VITRO [J].
DAILEY, ME ;
BUCHANAN, JA ;
BERGLES, DE ;
SMITH, SJ .
JOURNAL OF NEUROSCIENCE, 1994, 14 (03) :1060-1078
[4]   2-PHOTON LASER SCANNING FLUORESCENCE MICROSCOPY [J].
DENK, W ;
STRICKLER, JH ;
WEBB, WW .
SCIENCE, 1990, 248 (4951) :73-76
[5]  
HAY BA, 1994, DEVELOPMENT, V120, P2121
[6]   POWER AND LIMITS OF LASER-SCANNING CONFOCAL MICROSCOPY [J].
LAURENT, M ;
JOHANNIN, G ;
GILBERT, N ;
LUCAS, L ;
CASSIO, D ;
PETIT, PX ;
FLEURY, A .
BIOLOGY OF THE CELL, 1994, 80 (2-3) :229-240
[7]   THE JELLYFISH GREEN FLUORESCENT PROTEIN - A NEW TOOL FOR STUDYING ION-CHANNEL EXPRESSION AND FUNCTION [J].
MARSHALL, J ;
MOLLOY, R ;
MOSS, GWJ ;
HOWE, JR ;
HUGHES, TE .
NEURON, 1995, 14 (02) :211-215
[8]   MUTATIONS DISRUPTING NEURONAL CONNECTIVITY IN THE DROSOPHILA VISUAL-SYSTEM [J].
MARTIN, KA ;
POECK, B ;
ROTH, H ;
EBENS, AJ ;
BALLARD, LC ;
ZIPURSKY, SL .
NEURON, 1995, 14 (02) :229-240
[9]  
NAKAMURA O, 1993, OPTIK, V93, P39
[10]   DYNAMIC CHANGES IN OPTIC FIBER TERMINAL ARBORS LEAD TO RETINOTOPIC MAP FORMATION - AN INVIVO CONFOCAL MICROSCOPIC STUDY [J].
OROURKE, NA ;
FRASER, SE .
NEURON, 1990, 5 (02) :159-171