ANTISERUM TO LUCIFER YELLOW - PREPARATION, CHARACTERIZATION, AND USE FOR IMMUNOCYTOCHEMICAL LOCALIZATION OF DYE-FILLED RETINAL NEURONS

被引:6
作者
BRANDON, C [1 ]
CRISWELL, MH [1 ]
机构
[1] CHICAGO MED SCH,DEPT CELL BIOL & ANAT,3333 GREEN BAY RD,N CHICAGO,IL 60064
关键词
RABBIT; DOGFISH; RETINAL GANGLION CELLS; LUCIFER YELLOW; IMMUNOHISTOCHEMISTRY; ANTISERA;
D O I
10.1177/39.11.1918929
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
We present a new method for the preparation of antisera to Lucifer Yellow, and these antisera are here shown to be particularly suitable for immunocytochemical localization of multiple dye-injected cells in large pieces of vertebrate retina. The method involves the preparation of covalent conjugates of the VS isomer of Lucifer Yellow with keyhole limpet hemocyanin (KLH) or rabbit serum albumin (RSA), and their use as immunogens in rabbits. Both carrier protein conjugates yielded robust antibody responses. Antiserum to the KLH-LY conjugate contained precipitating antibodies against LY and KLH, although activity to the latter did not interfere with immunocytochemical staining. Rabbit antiserum to the RSA-LY conjugate contained precipitating antibody only against LY. When used for immunocytochemical staining of large retinal pieces containing many LY-filled cells, both antisera yielded well-stained, darkly filled cells similar to those seen with the Golgi technique; even very fine dendritic processes of retinal ganglion cells could be followed for long distances. LY immunocytochemistry provides a useful alternative to photooxidation for the analysis of multiple dye injected cells, especially in whole mounts. This approach may also be useful for immunocytochemical identification of cells filled with LY after tissue fixation.
引用
收藏
页码:1547 / 1553
页数:7
相关论文
共 17 条
[2]  
BOYD WC, 1956, FUNDAMENTALS IMMUNOL, P104
[4]   CHOLINERGIC AMACRINE NEURONS OF THE DOGFISH RETINA [J].
BRANDON, C .
VISUAL NEUROSCIENCE, 1991, 6 (06) :553-562
[5]   BURST GENERATING AND REGULAR SPIKING LAYER-5 PYRAMIDAL NEURONS OF RAT NEOCORTEX HAVE DIFFERENT MORPHOLOGICAL FEATURES [J].
CHAGNACAMITAL, Y ;
LUHMANN, HJ ;
PRINCE, DA .
JOURNAL OF COMPARATIVE NEUROLOGY, 1990, 296 (04) :598-613
[6]  
HASHIDA S, 1984, Journal of Applied Biochemistry, V6, P56
[7]  
Haugland R P., 1989, HDB FLUORESCENT PROB
[8]   A VERSATILE MEANS OF INTRACELLULAR LABELING - INJECTION OF BIOCYTIN AND ITS DETECTION WITH AVIDIN CONJUGATES [J].
HORIKAWA, K ;
ARMSTRONG, WE .
JOURNAL OF NEUROSCIENCE METHODS, 1988, 25 (01) :1-11
[10]   PHOTOCONVERSION OF SOME FLUORESCENT MARKERS TO A DIAMINOBENZIDINE PRODUCT [J].
SANDELL, JH ;
MASLAND, RH .
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 1988, 36 (05) :555-559