A method for direct imaging of x-z cross-sections of fluorescent samples.
A method for direct imaging of x-z cross-sections of fluorescent samples.
复制标题
荧光样品x-z截面的直接成像方法。
DOI：
10.1111/jmi.12965
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发表时间：
2021-03
影响因子：
2
通讯作者：
Lele TP
中科院分区：
文献类型：
--
作者：
Katiyar A;Antani JD;McKee BP;Gupta R;Lele PP;Lele TP
荧光物体的x-z横截面轮廓在共焦显微镜中可能会失真，这在很大程度上是由于典型的高数值孔径物镜的浸没介质的折射率与其中存在样品的介质的折射率之间的失配。在这里，我们介绍了一种方法，安装荧光样品平行于光轴。这种安装允许在显微镜的x-y平面中直接成像通常是物体的x-z横截面。通过这种方法，与从共焦z堆叠重建的x-z横截面相比，荧光珠的x-y横截面被认为具有显著更低的形状失真。我们进一步测试了培养细胞中细胞核和细胞高度成像的方法，发现它们比以前报道的要平坦得多。该方法允许荧光样品的x-z横截面的改进的成像。
The x-z cross-sectional profiles of fluorescent objects can be distorted in confocal microscopy, in large part due to mismatch between the refractive index of the immersion medium of typical high numerical aperture objectives and the refractive index of the medium in which the sample is present. Here, we introduce a method to mount fluorescent samples parallel to the optical axis. This mounting allows direct imaging of what would normally be an x-z cross-section of the object, in the x-y plane of the microscope. With this approach, the x-y cross-sections of fluorescent beads were seen to have significantly lower shape-distortions as compared to x-z cross-sections reconstructed from confocal z-stacks. We further tested the method for imaging of nuclear and cellular heights in cultured cells, and found that they are significantly flatter than previously reported. This approach allows improved imaging of the x-z cross-section of fluorescent samples.
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