Microwaves and heat in aldehyde fixation: Model experiments with bovine serum albumin

被引:4
作者
Hopwood, D [1 ]
机构
[1] Univ Dundee, Ninewells Hosp & Med Sch, Dept Pathol, Gastroenterol Lab, Dundee DD1 9SY, Scotland
关键词
D O I
10.1006/meth.1998.0614
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Most model experiments concerning tissue fixation have used low concentrations of fixatives and proteins. Here, high concentrations (up to 32%) of bovine serum albumin (BSA) were reacted with formaldehyde (1-20%) and glutaraldehyde (0.5-4%). Gels were formed between 16% BSA and 10-20% formaldehyde at room temperature, but not with percentages of formaldehyde lower than 4%. Microwave application or heating in a water bath to 50 degrees C gave a gel from 1 to 20% formaldehyde. Sixteen percent BSA also gave a gel with glutaraldehyde from 0.5 to 4%. Cone and plate viscometry showed rapidly increasing viscosity at 4% formaldehyde and 16% BSA at room temperature. At 50 degrees C, gels formed at a low concentration of formaldehyde. Tissue fixation in which the local concentrations of protein may be in excess of 30% is probably more complete than in vitro experiments in which low concentrations of reagents have been used to permit subsequent spectrometry. This was confirmed by polyacrylamide gel electrophoresis of liver, (C) 1998 Academic Press.
引用
收藏
页码:119 / 122
页数:4
相关论文
共 15 条
[1]   PRESERVATION OF MYELINATED FIBERS FOR ELECTRON-MICROSCOPY - A QUALITATIVE COMPARISON OF ALDEHYDE FIXATION, MICROWAVE STABILIZATION AND OTHER PROCEDURES ALL COMPLETED BY OSMICATION [J].
FEIRABEND, HKP ;
KOK, P ;
CHOUFOER, H ;
PLOEGER, S .
JOURNAL OF NEUROSCIENCE METHODS, 1994, 55 (02) :137-153
[2]  
FEIRABEND HKP, 1991, EUR J MORPHOL, V29, P185
[3]   A COMPARISON OF CROSSLINKING ABILITIES OF GLUTARALDEHYDE FORMALDEHYDE AND ALPHA-HYDROXYADIPALDEHYDE WITH BOVINE SERUM ALBUMIN AND CASEIN [J].
HOPWOOD, D .
HISTOCHEMIE, 1969, 17 (02) :151-+
[4]   DIFFERENTIATING THE EFFECTS OF MICROWAVE AND HEAT ON TISSUE PROTEINS AND THEIR CROSSLINKING BY FORMALDEHYDE [J].
HOPWOOD, D ;
YEAMAN, G ;
MILNE, G .
HISTOCHEMICAL JOURNAL, 1988, 20 (6-7) :341-346
[5]   MICROWAVE FIXATION - ITS POTENTIAL FOR ROUTINE TECHNIQUES, HISTOCHEMISTRY, IMMUNOCYTOCHEMISTRY AND ELECTRON-MICROSCOPY [J].
HOPWOOD, D ;
COGHILL, G ;
RAMSAY, J ;
MILNE, G ;
KERR, M .
HISTOCHEMICAL JOURNAL, 1984, 16 (11) :1171-1191
[6]  
HOPWOOD D, 1991, ELECT MICROSCOPY BIO, P1
[7]  
Kok L. P., 1992, MICROWAVE COOKBOOK M
[8]   CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].
LAEMMLI, UK .
NATURE, 1970, 227 (5259) :680-+
[9]   ANTIGEN PRESERVATION IN MICROWAVE-IRRADIATED TISSUES - A COMPARISON WITH FORMALDEHYDE FIXATION [J].
LEONG, ASY ;
MILIOS, J ;
DUNCIS, CG .
JOURNAL OF PATHOLOGY, 1988, 156 (04) :275-282
[10]   FIXATION FOR BIOLOGICAL ULTRASTRUCTURE .1. A VISCOMETRIC ANALYSIS OF THE INTERACTION BETWEEN GLUTARALDEHYDE AND BOVINE SERUM-ALBUMIN [J].
LUFT, JH .
JOURNAL OF MICROSCOPY, 1992, 167 :247-258