Adler H J, Mantooth J, Raphael Y
Kresge Hearing Research Institute, Ann Arbor, MI 48109, USA.
Scanning Microsc. 1995 Sep;9(3):825-30; discussion 830-1.
Neonatal chicks were exposed to an octave band noise with a center frequency of 1.5 kHz at 116 dB SPL for 4 hours. Seven days following overstimulation, the birds were sacrificed. Their basilar papillae were removed, fixed in 4% paraformaldehyde, and processed in two steps. First, the ears were immunostained with a supernatant of mouse anti-tectorial membrane antibodies, followed by a diaminobenzidine process. Examinations of the papillae under an optical stereo microscope revealed a patch site with a partially regenerated tectorial membrane (referred to as the honeycomb). After the optical studies, the same ears were post-fixed in 1% osmium tetroxide, dehydrated in ethanol, and processed for scanning electron microscopy (SEM). SEM examinations demonstrated a honeycomb-covered patch lesion in the papilla. Patch lesion perimeters were traced from both the optical and SEM images, and patch areas were calculated. Also, papilla height was measured at the midpoint of the inner ear in both groups. These calculations showed that the patch area and papilla height had shrunk by approximately 37% and 33%, respectively, following the SEM methodology. The decrease in these dimensions may be attributed to several steps required for the SEM specimen preparation, such as critical point drying.
将新生雏鸡暴露于中心频率为1.5 kHz、声压级为116 dB SPL的倍频程带噪声中4小时。过度刺激7天后，处死这些雏鸡。取出它们的基底乳头，固定在4%多聚甲醛中，并分两步进行处理。首先，用小鼠抗盖膜抗体的上清液对耳朵进行免疫染色，然后进行二氨基联苯胺处理。在光学立体显微镜下检查乳头，发现一个盖膜部分再生的斑块部位（称为蜂窝状）。光学研究后，将同一只耳朵再固定在1%四氧化锇中，用乙醇脱水，并进行扫描电子显微镜（SEM）处理。SEM检查显示乳头中有一个被蜂窝状覆盖的斑块病变。从光学图像和SEM图像中追踪斑块病变的周长，并计算斑块面积。此外，在两组内耳的中点测量乳头高度。这些计算表明，按照SEM方法，斑块面积和乳头高度分别缩小了约37%和33%。这些尺寸的减小可能归因于SEM标本制备所需的几个步骤，如临界点干燥。
