Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
Appl Spectrosc. 2013 Jan;67(1):93-105. doi: 10.1366/11-06568.
The quality of images from an infrared (IR) microscope has traditionally been limited by considerations of throughput and signal-to-noise ratio (SNR). An understanding of the achievable quality as a function of instrument parameters, from first principals is needed for improved instrument design. Here, we first present a model for light propagation through an IR spectroscopic imaging system based on scalar wave theory. The model analytically describes the propagation of light along the entire beam path from the source to the detector. The effect of various optical elements and the sample in the microscope is understood in terms of the accessible spatial frequencies by using a Fourier optics approach and simulations are conducted to gain insights into spectroscopic image formation. The optimal pixel size at the sample plane is calculated and shown much smaller than that in current mid-IR microscopy systems. A commercial imaging system is modified, and experimental data are presented to demonstrate the validity of the developed model. Building on this validated theoretical foundation, an optimal sampling configuration is set up. Acquired data were of high spatial quality but, as expected, of poorer SNR. Signal processing approaches were implemented to improve the spectral SNR. The resulting data demonstrated the ability to perform high-definition IR imaging in the laboratory by using minimally-modified commercial instruments.
传统上,红外(IR)显微镜的图像质量受到吞吐量和信噪比(SNR)的考虑因素限制。为了改进仪器设计,需要从第一性原理上了解仪器参数作为函数的可实现质量。在这里,我们首先根据标量波动理论,为基于光谱学的成像系统的光传播建立了一个模型。该模型从理论上描述了光从光源到探测器的整个光束路径的传播。通过傅里叶光学方法,可以根据可用的空间频率来理解显微镜中各种光学元件和样本的作用,并进行模拟以深入了解光谱成像的形成。计算了样品平面上的最佳像素尺寸,并表明其远小于当前中红外显微镜系统中的像素尺寸。对商业成像系统进行了修改,并给出了实验数据以验证所开发模型的有效性。在此经过验证的理论基础上,建立了最佳采样配置。所获取的数据具有较高的空间质量,但如预期的那样,SNR 较差。实施了信号处理方法来提高光谱 SNR。所得数据证明了仅使用最小修改的商用仪器即可在实验室中进行高清晰度 IR 成像的能力。