A rapid microbiopsy system to improve the preservation of biological samples prior to high-pressure freezing

被引:25
作者
Vanhecke, D
Graber, W
Herrmann, G
Al-Amoudi, A
Eggli, P
Studer, D [1 ]
机构
[1] Univ Bern, Inst Anat, Bern, Switzerland
[2] Univ Lausanne, Lab Anal Ultrastruct, Lausanne, Switzerland
关键词
cryofixation; cryosectioning; electron microscopy; freeze-substitution; high-pressure freezing; microbiopsy;
D O I
10.1046/j.1365-2818.2003.01226.x
中图分类号
TH742 [显微镜];
学科分类号
摘要
A microbiopsy system for fast excision and transfer of biological specimens from donor to high-pressure freezer was developed. With a modified, commercially available, Promag 1.2 biopsy gun, tissue samples can be excised with a size small enough (0.6 mm x 1.2 mm x 0.3 mm) to be easily transferred into a newly designed specimen platelet. A self-made transfer unit allows fast transfer of the specimen from the needle into the specimen platelet. The platelet is then fixed in a commercially available specimen holder of a high-pressure freezing machine (EM PACT, Leica Microsystems, Vienna, Austria) and frozen therein. The time required by a well-instructed (but not experienced) person to execute all steps is in the range of half a minute. This period is considered short enough to maintain the excised tissue pieces close to their native state. We show that a range of animal tissues (liver, brain, kidney and muscle) are well preserved. To prove the quality of freezing achieved with the system, we show vitrified ivy leaves high-pressure frozen in the new specimen platelet.
引用
收藏
页码:3 / 12
页数:10
相关论文
共 31 条
[11]   DIAMONDS ARE A CRYOSECTIONERS BEST FRIEND [J].
MICHEL, M ;
GNAGI, H ;
MULLER, M .
JOURNAL OF MICROSCOPY, 1992, 166 :43-56
[12]  
Monaghan P, 1998, J MICROSC-OXFORD, V192, P248, DOI 10.1046/j.1365-2818.1998.00387.x
[13]  
Moor H, 1987, Cryotechniques in biological electron microscopy, P175, DOI [10.1007/978-3-642-72815-0_8, DOI 10.1007/978-3-642-72815-0_8]
[14]  
MULLER M, 1980, MIKROSKOPIE, V36, P129
[15]  
Muller M., 1984, SCI BIOL SPECIMEN PR, P131
[16]   HIGH-DENSITY MORPHOLOGIES OF ICE IN HIGH-PRESSURE FROZEN BIOLOGICAL SPECIMENS [J].
RICHTER, K .
ULTRAMICROSCOPY, 1994, 53 (03) :237-249
[17]   CUTTING ARTIFACTS ON ULTRATHIN CRYOSECTIONS OF BIOLOGICAL BULK SPECIMENS [J].
RICHTER, K .
MICRON, 1994, 25 (04) :297-308
[18]  
Royer SM, 1996, MICROSC RES TECHNIQ, V35, P385
[19]   STRUCTURAL-CHANGES OF DUODENUM BRUSH-BORDER IN LUNG-CANCER PATIENTS TREATED WITH CISPLATIN PLUS ETOPOSIDE [J].
SARTORI, S ;
NIELSEN, I ;
TREVISANI, L ;
TASSINARI, D ;
ALEOTTI, A ;
GRANDI, E ;
ABBASCIANO, V .
LUNG CANCER, 1993, 10 (1-2) :55-62
[20]  
Shimoni E, 1998, J MICROSC-OXFORD, V192, P236, DOI 10.1046/j.1365-2818.1998.00389.x