参数 | 推荐起点 | 调整依据 | 常见错误 |
|---|---|---|---|
物镜/工作距离 | 水镜或长工作距离物镜,WD > 0.6 mm | 球体直径与容器壁厚 [1][3] | 用高 NA 油镜硬拍厚样本 |
校正环 | 按实际盖玻片/壁厚设定 | 深层边缘发虚、对比度下降 [4] | 始终停在出厂位置 |
针孔 | 1 AU(需要更薄切片时 0.7 AU) | 光学切片厚度与信噪比 [2] | 为提亮把针孔开到 2 AU 以上 |
z 步距 | 1 µm,且 ≤ 轴向分辨率一半 | 轴向分辨率 2nλ/NA² [5] | 步距 3–5 µm 后做三维重建 |
主增益 | ≤ 800–850 | 范围指示器无饱和 [2] | 靠拉增益代替延长曝光 |
数字增益 | 保持 1 | 避免噪声被同倍放大 [2] | 用数字增益"提亮" |
位深/格式 | 12 或 16 位 | 定量与灰度层次 [2] | 8 位采集后做强度比较 |
z 轴补偿 | 由上而下线性递增功率/增益 | 深层信号衰减 [1] | 全程固定功率 |
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