High Magnification Z-Stacks from Blood Films

All biological samples were collected from participants recruited under the auspices of the CMRG at the 800-bed tertiary hospital, UCH in the city of Ibadan, Nigeria, after after de-identification of the patient information. We trained and tested the CAMI-Fusion network on three different types of samples: (a) Giemsa-stained thick blood films (TBF) and (b) Giemsa-stained peripheral blood smears (PBS) for malaria parasite detection as well as (c) Wright-stained Bone Marrow Aspirates (BMA) for diagnosing blood cancers.<i><br></i><i>Image acquisition and pre-processing:</i> We used an upright brightfield microscope (BX63, Olympus, 100W halogen bulb light source) with a 100X/1.4NA oil immersion objective (MPlanApoN, Olympus) and a color digital camera (Edge 5.5C, PCO) to acquire multiple high-resolution z-stacks with an axial step of a 0.5 µm. White balancing was applied to each focal plane image. For each sample acquisition, an empty field of view (empty region of the slide not containing any blood cells) was acquired and considered as a white object reference.

所有生物样本均采集自尼日利亚伊巴丹市拥有800张床位的三级医院UCH，由CMRG主持招募的受试者，且已完成患者信息的去标识化处理。我们针对三类不同样本开展了CAMI-Fusion网络的训练与测试：(a) 吉姆萨染色厚血膜（Thick Blood Film, TBF）与(b) 吉姆萨染色外周血涂片（Peripheral Blood Smear, PBS），二者均用于疟原虫检测；(c) 瑞氏染色骨髓穿刺物（Bone Marrow Aspirates, BMA），用于血液癌症诊断。<br>图像采集与预处理：我们采用正置明场显微镜（upright brightfield microscope，型号BX63，奥林巴斯，100W卤素灯光源）搭配100X/1.4NA油浸物镜（oil immersion objective，型号MPlanApoN，奥林巴斯）及彩色数码相机（Edge 5.5C，PCO），采集轴向步长为0.5 μm的多幅高分辨率Z轴堆叠图像（z-stacks）。我们对每个焦平面图像均执行白平衡处理。在每次样本采集过程中，我们还会采集一片空白视野（即载玻片上不含任何血细胞的区域），将其作为白色参考对象。