Multi-photon microscopy of cell types in the viable taste disk of the frog
Multi-photon microscopy of cell types in the viable taste disk of the frog
复制标题
青蛙活味盘中细胞类型的多光子显微镜
DOI：
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复制
发表时间：
2003
影响因子：
3.6
通讯作者：
B. Lindemann
中科院分区：
文献类型：
--
作者：
Jack H. Y. Li;B. Lindemann
用多光子显微镜观察了蛙的活味盘的形态。为了鉴定组织内的单个感觉或支持细胞，我们寻找对细胞群或可能的细胞类型的亚群染色的荧光染料。某些细胞类型确实优先用某些荧光染料染色。一个亚组的胶质样细胞（Ic型）染色BCECF，H+敏感染料，和indo-1，Ca 2+敏感染料，都在膜渗透酯的形式。BCECF-酯也染色III型受体细胞的树突，但indo-1酯没有。II型受体细胞用MQAE染色，MQAE是一种带正电荷的Cl−敏感染料。一个亚群的II型细胞积累阿米洛利，带正电荷的荧光利尿剂。某些支持细胞，即，翼细胞（Ib型）和神经胶质样细胞（Ic型），用带负电荷的染料标记，例如，钙绿色-1葡聚糖。粘液细胞（Ia型）染色的19种染料中只有两种检查，和梅克尔样基底细胞（IV型）只染色膜标记的电压敏感染料，推测通过内吞作用。未发现可染色所有类型细胞或所有受体细胞的染料。这一发现揭示了未来针对群体反应的功能成像的潜在问题，因为未染色细胞的反应将保持未观察到。染料对细胞类型的特异性足以识别支持细胞和受体细胞。然后可以使用光学切片和渲染技术重建细胞形状。因此，装载染料的细长细胞群，特别是Ic、II和III型细胞，可以首次在三维中可视化。
The morphology of viable taste disks of the frog was explored with multi-photon microscopy. In order to identify single sensory or supporting cells within the tissue, we searched for fluorescent dyes that stained subsets of the cell population or possibly cell types. Some cell types indeed stained preferentially with certain fluorescent dyes. A subset of glia-like cells (type Ic) stained with BCECF, a H+-sensitive dye, and indo-1, a Ca2+-sensitive dye, both presented in the membrane-permeant ester form. BCECF-ester also stained the dendrites of type III receptor cells, but indo-1 ester did not. Receptor cells of type II stained with MQAE, a positively charged Cl−-sensitive dye. A subset of type II cells accumulated amiloride, a positively charged fluorescent diuretic. Certain supporting cells, i.e., wing cells (type Ib) and glia-like cells (type Ic), were labeled by negatively charged dyes, e.g., calcium green-1 dextran. Mucus cells (type Ia) were stained with only two of the 19 dyes examined, and Merkel-like basal cells (type IV) were stained only with a membrane-labeling voltage-sensitive dye, presumably by endocytosis. No dye was found which would stain all types of cells or all receptor cells. This finding reveals a potential problem for future functional imaging aiming at population responses, as the responses of unstained cells then would remain unobserved. Specificity of dyes with respect to cell types was sufficient to identify supporting cells and receptor cells. Cell shape could then be reconstructed, using optical slicing and rendering techniques. Thus populations of dye-loaded elongated cells, especially types Ic, II and III, could for the first time be visualized in three dimensions.
影响因子：
3.4
作者：
NICHOLSON, C;TAO, L
通讯作者：
TAO, L