Departamento de Óptica y de Optometría y Ciencias de la Visión, Facultad de Física, Universidad de Valencia, C/Doctor Moliner 50, 46100 Burjassot, Spain.
Biosensors (Basel). 2023 Feb 10;13(2):253. doi: 10.3390/bios13020253.
The combination of different imaging modalities into single imaging platforms has a strong potential in biomedical sciences as it permits the analysis of complementary properties of the target sample. Here, we report on an extremely simple, cost-effective, and compact microscope platform for achieving simultaneous fluorescence and quantitative phase imaging modes with the capability of working in a single snapshot. It is based on the use of a single illumination wavelength to both excite the sample's fluorescence and provide coherent illumination for phase imaging. After passing the microscope layout, the two imaging paths are separated using a bandpass filter, and the two imaging modes are simultaneously obtained using two digital cameras. We first present calibration and analysis of both fluorescence and phase imaging modalities working independently and, later on, experimental validation for the proposed common-path dual-mode imaging platform considering static (resolution test targets, fluorescent micro-beads, and water-suspended lab-made cultures) as well as dynamic (flowing fluorescent beads, human sperm cells, and live specimens from lab-made cultures) samples.
将不同的成像模式组合到单个成像平台中在生物医学科学中有很大的潜力,因为它允许分析目标样本的互补特性。在这里,我们报告了一种极其简单、经济高效且紧凑的显微镜平台,用于实现具有单次拍摄能力的荧光和定量相位成像模式的同时采集。它基于使用单个照明波长同时激发样品的荧光并为相位成像提供相干照明。经过显微镜布局后,使用带通滤波器分离两个成像路径,并使用两个数字相机同时获得两种成像模式。我们首先介绍了独立工作的荧光和相位成像模式的校准和分析,然后基于静态(分辨率测试目标、荧光微珠和水悬浮的实验室培养物)和动态(流动荧光珠、人类精子细胞和实验室培养物的活标本)样本对所提出的共路双模成像平台进行了实验验证。