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Dumont #5SF forceps (super-fine)

Company: WPI
Catalog#: 500085
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Preparation of Precisely Oriented Cryosections of Undistorted Drosophila Wing Imaginal Discs for High Resolution Confocal Imaging
[Abstract]  The combination of immunofluorescence and laser scanning confocal microscopy (LSM) is essential to high-resolution detection of molecular distribution in biological specimens. A frequent limitation is the need to image deep inside a tissue or in a specific plane, which may be inaccessible due to tissue size or shape. Recreating high-resolution 3D images is not possible because the point-spread function of light reduces the resolution in the Z-axis about 3-fold, compared to XY, and light scattering obscures signal deep in the tissue. However, the XY plane of interest can be chosen if embedded samples are precisely oriented and sectioned prior to imaging (Figure 1). Here we describe the preparation of frozen tissue sections of the Drosophila wing imaginal disc, which allows us to ... [摘要]  免疫荧光和激光扫描共聚焦显微镜(LSM)的组合是高分辨率检测生物样品中分子分布的关键。频繁的限制是需要在组织内或在特定的平面深处进行成像,这可能由于组织大小或形状而不可接近。因为与XY相比,光的点扩散函数将Z轴的分辨率降低了约3倍,并且光散射使组织中的深层信号模糊,所以不可能重新创建高分辨率3D图像。然而,如果嵌入的样品在成像之前被精确地定向和切片,则可以选择感兴趣的XY平面(图1)。在这里,我们描述的果蝇翅成像光盘的冰冻组织切片的准备,这使得我们能够获得高分辨率的图像,在整个这个折叠上皮的深度。

图1.上皮结构和未畸变的折叠模式在发育果蝇翅膀的这个冰冻部分的整个深度中都被揭示出来。通过机翼囊横向背腹节。 A.冷冻切片显示贯穿上皮深度的信号的α-连环蛋白(A',A“,洋红色)的细胞核(A,绿色)和亚细胞分布。基底表面清晰可辨(箭头)。 A是“A的数字增强图像”。 B.在显示为XZ正交视图的自顶向下视图中收集的图像的Z-堆叠揭示了α-连环蛋白(B',B“)甚至数字增强图像(B”)的细胞核(B)但很少可辨别的细节。未能揭示基底上皮表面(箭头)。 ...