Localization of GFP in frozen sections from unfixed mouse tissues: Immobilization of a highly soluble marker protein by formaldehyde vapor

被引:51
作者
Jockusch, H [1 ]
Voigt, S [1 ]
Eberhard, D [1 ]
机构
[1] Univ Bielefeld, D-33501 Bielefeld, Germany
关键词
green fluorescent protein; transgenes; vapor fixation;
D O I
10.1177/002215540305100315
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Green fluorescent protein (GFP) and its variants, such as enhanced GFP (EGFP), have been introduced into mammalian cells by transgenes, e.g., to distinguish donor from host cells after transplantation. Free GFP is extremely soluble and leaks out from liquid-covered cryostat sections so that fixation of whole organs before sectioning has been mandatory. This precludes the analysis of serial sections with respect to fixation-sensitive enzyme activities and antigens. We describe here a vapor fixation for sections from unfixed cryostat blocks of tissue that allows unrestricted enzyme and immunohistochemistry on adjacent sections, as demonstrated for cross-striated muscle and other tissues from EGFP transgenic "green mice" and for a transplantation experiment.
引用
收藏
页码:401 / 404
页数:4
相关论文
共 7 条
[1]   IMMUNOCHEMICAL ANALYSIS OF MYOSIN HEAVY-CHAIN DURING AVIAN MYOGENESIS INVIVO AND INVITRO [J].
BADER, D ;
MASAKI, T ;
FISCHMAN, DA .
JOURNAL OF CELL BIOLOGY, 1982, 95 (03) :763-770
[2]  
Eberhard D., 2002, J MUSCLE RES CELL M, V23, P175
[3]   DIRECT CORRELATION OF PARVALBUMIN LEVELS WITH MYOSIN ISOFORMS AND SUCCINATE-DEHYDROGENASE ACTIVITY ON FROZEN-SECTIONS OF RODENT MUSCLE [J].
FUCHTBAUER, EM ;
ROWLERSON, AM ;
GOTZ, K ;
FRIEDRICH, G ;
MABUCHI, K ;
GERGELY, J ;
JOCKUSCH, H .
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 1991, 39 (03) :355-361
[4]  
JOCKUSCH H, 2000, J MUSCLE RES CELL M, V21, P799
[5]   CYTOCHEMICAL DEMONSTRATION OF SUCCINIC DEHYDROGENASE BY THE USE OF A NEW PARA-NITROPHENYL SUBSTITUTED DITETRAZOLE [J].
NACHLAS, MM ;
TSOU, KC ;
DESOUZA, E ;
CHENG, CS ;
SELIGMAN, AM .
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 1957, 5 (04) :420-436
[6]   'Green mice' as a source of ubiquitous green cells [J].
Okabe, M ;
Ikawa, M ;
Kominami, K ;
Nakanishi, T ;
Nishimune, Y .
FEBS LETTERS, 1997, 407 (03) :313-319
[7]   Rapid and sensitive detection of enhanced green fluorescent protein expression in paraffin sections by confocal laser scanning microscopy [J].
Walter, I ;
Fleischmann, M ;
Klein, D ;
Müller, M ;
Salmons, B ;
Günzburg, WH ;
Renner, M ;
Gelbmann, W .
HISTOCHEMICAL JOURNAL, 2000, 32 (02) :99-103