![[Translate to chinese:] Cell counts for each biomarker were divided by total number of cells to give a percentage of biomarker positive cells out of total cells for each biomarker. Cell_counts_for_biomarkers_with_Cell_DIVE.jpg](/fileadmin/_processed_/5/3/csm_Cell_counts_for_biomarkers_with_Cell_DIVE_8a0d1d805f.jpg "[Translate to chinese:] Cell counts for each biomarker were divided by total number of cells to give a percentage of biomarker positive cells out of total cells for each biomarker.")
关于案例研究
可重复性是科学研究领域挥之不去的危机，空间生物学研究也不例外。多个因素会影响实验结果的可重复性。抗体验证是癌症研究中的重要步骤，以确保所用抗体具有特异性和可靠性。这些抗体必须能在不同批次和时间的样品实现稳定的染色。近年来，癌症研究越来越多地采用自动化系统，以提高实验的可重复性和效率。
此案例研究中，我们测试了Cell DIVE多重成像解决方案的可重复性。Cell DIVE装备了来自Advanced Solutions Life Sciences（ASLS）的BAB 200自动化系统。我们使用来自Cell Signaling Technology（CST）的验证抗体。Cell DIVE可以在整个组织切片上探测到数十种生物标志物并进行成像。这通过迭代染色和染料灭活工作流程完成。本质上，Cell DIVE致力于提供实验方法的可重复性，从组织制备、抗原修复到样品成像和载玻片储存。Cell DIVE可以使用直接偶联的一抗，而这是变异性的另一个来源。
令人欣慰的是，来自CST的偶联抗体经过严格验证，可以减少变异性。此外，重组抗体的使用、一致的偶联物亮度和抗体标记度可以降低不同批次之间的变异性，确保了空间生物学研究所需的可靠偶联抗体。高分辨率的成像结果源于一致的轮间成像、稳定的校准以及使用ASLS的BAB 200液体处理解决方案减少人为错误。我们还使用以不同批次CST抗体组合探测的相邻组织切片。然后，使用配备BAB 200液体处理器的Cell DIV成像仪，按照时间分批成像。经过可靠性分析，我们报告了不同参数下的可重复性结果。空间生物学的研究方法和实验结果的可重复性对于获取可靠的洞察信息至关重要，有助于让人们对旨在改善患者预后的未来研究的质量充满信心。
![[Translate to chinese:] Multiplexed Cell DIVE imaging of Adult Human Alzheimer’s brain tissue section demonstrating expression of markers specific to astrocytes (GFAP, S100B), microglia (TMEM119, IBA1), AD-associated markers (p-Tau217, β-amyloid) and immune cells such as CD11b+, CD163+, CD4+, and HLA-DRA+, clustered around the β-amyloid plaques.](/fileadmin/_processed_/b/b/csm_Alzheimers_brain_tissue_section_showing_astrocytes_microglia_immune_cells_26ab13cde2.jpg "[Translate to chinese:] Multiplexed Cell DIVE imaging of Adult Human Alzheimer’s brain tissue section demonstrating expression of markers specific to astrocytes , microglia , AD-associated markers and immune cells clustered around the β-amyloid plaques")
![[Translate to chinese:] Multiplexed Cell DIVE imaging to characterize the spatial landscape in Human Alzheimer’s Cortical Tissue](/fileadmin/_processed_/7/2/csm_Human_Alzheimers_Cortical_Tissue_Multiplexed_Cell_DIVE_imaging_b053b78344.jpg "[Translate to chinese:] Multiplexed Cell DIVE imaging to characterize the spatial landscape in Human Alzheimer’s Cortical Tissue")
