Comparing and Correcting Spectral Sensitivities between Multispectral Microscopes: A Prerequisite to Clinical Implementation.
Comparing and Correcting Spectral Sensitivities between Multispectral Microscopes: A Prerequisite to Clinical Implementation.
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DOI：
10.3390/cancers15123109
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发表时间：
2023-06-08
期刊：
影响因子：
5.2
通讯作者：
Szalay, Alexander S.
中科院分区：
文献类型：
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作者：
Eminizer, Margaret;Nagy, Melinda;Engle, Elizabeth L.;Soto-Diaz, Sigfredo;Jorquera, Andrew;Roskes, Jeffrey S.;Green, Benjamin F.;Wilton, Richard;Taube, Janis M.;Szalay, Alexander S.
多光谱、多重免疫荧光（mIF）显微镜是一种表征肿瘤微环境的新兴技术。实现mIF显微图像的高通量收集和分析通常需要使用多个显微镜，但不能保证来自一个显微镜的数据可以与来自另一个显微镜的数据进行比较。我们使用了一组八个黑色素瘤组织样本来测量和校正三个显微镜之间的数据差异。我们在每个显微镜上扫描样品两次，并测量所得图像组中的平均组织通量密度。通过应用一个相对简单的校准模型，考虑到样品和显微镜特定的影响，我们能够减少79%和72%，分别在原始图像亮度和免疫标记物表达测量的变化。这表明，简单的程序可以用来有效地标准化mIF数据从多个显微镜在研究和临床诊断设置的潜在用途。多光谱、多重免疫荧光（mIF）显微镜已在研究中发挥了巨大作用，以识别组织内的细胞共表达谱和空间关系，提供了无数的诊断优势。随着这些技术的成熟，来自mIF显微镜的图像数据在设备上的可再现性和可扩展性至关重要。我们试图表征和校正三个多光谱显微镜之间的照明强度和光谱灵敏度的差异。我们在每个显微镜上扫描八个黑素瘤组织样本两次，并计算其平均组织区域通量强度。我们发现所有显微镜、扫描和样本的基线平均标准差为29.9%，在应用样本特定校正后，该标准差降低至13.9%，说明了每张载玻片上显示的组织差异。我们使用基本校准模型来纠正样本和显微镜特定对整体亮度和相对亮度（作为图像层的函数）的影响。我们测试了校准程序的普遍性，发现对样本的独立验证子集应用校正将变异降低到2.9 ± 0.03%。通过将原始图像校正到单个参考显微镜，未混合标记表达的变化从15.8%减少到4.4%。我们的研究结果表明，MIF显微镜可以使用一个相对简单的校正模型标准化用于临床病理学实验室。
Multispectral, multiplex immunofluorescence (mIF) microscopy is an emerging technology for characterization of the tumour microenvironment. Achieving high-throughput collection and analysis of mIF microscopy images often requires the use of multiple microscopes, but it is not guaranteed that data from one microscope can be compared to data from another. We used a set of eight melanoma tissue samples to measure and correct for data differences between three microscopes. We scanned the samples twice on each microscope and measured the average tissue flux densities in the resulting sets of images. By applying a relatively simple calibration model accounting for sample- and microscope-specific effects, we were able to reduce the variations in raw image brightness and immune marker expression measurements by 79% and 72%, respectively. This shows that simple procedures can be used to effectively standardize mIF data from multiple microscopes for potential use in both research and clinical diagnostic settings. Multispectral, multiplex immunofluorescence (mIF) microscopy has been used to great effect in research to identify cellular co-expression profiles and spatial relationships within tissue, providing a myriad of diagnostic advantages. As these technologies mature, it is essential that image data from mIF microscopes is reproducible and standardizable across devices. We sought to characterize and correct differences in illumination intensity and spectral sensitivity between three multispectral microscopes. We scanned eight melanoma tissue samples twice on each microscope and calculated their average tissue region flux intensities. We found a baseline average standard deviation of 29.9% across all microscopes, scans, and samples, which was reduced to 13.9% after applying sample-specific corrections accounting for differences in the tissue shown on each slide. We used a basic calibration model to correct sample- and microscope-specific effects on overall brightness and relative brightness as a function of the image layer. We tested the generalizability of the calibration procedure and found that applying corrections to independent validation subsets of the samples reduced the variation to 2.9 ± 0.03%. Variations in the unmixed marker expressions were reduced from 15.8% to 4.4% by correcting the raw images to a single reference microscope. Our findings show that mIF microscopes can be standardized for use in clinical pathology laboratories using a relatively simple correction model.
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影响因子：
64.8
作者：
Helmink BA;Reddy SM;Gao J;Zhang S;Basar R;Thakur R;Yizhak K;Sade-Feldman M;Blando J;Han G;Gopalakrishnan V;Xi Y;Zhao H;Amaria RN;Tawbi HA;Cogdill AP;Liu W;LeBleu VS;Kugeratski FG;Patel S;Davies MA;Hwu P;Lee JE;Gershenwald JE;Lucci A;Arora R;Woodman S;Keung EZ;Gaudreau PO;Reuben A;Spencer CN;Burton EM;Haydu LE;Lazar AJ;Zapassodi R;Hudgens CW;Ledesma DA;Ong S;Bailey M;Warren S;Rao D;Krijgsman O;Rozeman EA;Peeper D;Blank CU;Schumacher TN;Butterfield LH;Zelazowska MA;McBride KM;Kalluri R;Allison J;Petitprez F;Fridman WH;Sautès-Fridman C;Hacohen N;Rezvani K;Sharma P;Tetzlaff MT;Wang L;Wargo JA
通讯作者：
Wargo JA
影响因子：
28.2
作者：
Chen PL;Roh W;Reuben A;Cooper ZA;Spencer CN;Prieto PA;Miller JP;Bassett RL;Gopalakrishnan V;Wani K;De Macedo MP;Austin-Breneman JL;Jiang H;Chang Q;Reddy SM;Chen WS;Tetzlaff MT;Broaddus RJ;Davies MA;Gershenwald JE;Haydu L;Lazar AJ;Patel SP;Hwu P;Hwu WJ;Diab A;Glitza IC;Woodman SE;Vence LM;Wistuba II;Amaria RN;Kwong LN;Prieto V;Davis RE;Ma W;Overwijk WW;Sharpe AH;Hu J;Futreal PA;Blando J;Sharma P;Allison JP;Chin L;Wargo JA
通讯作者：
Wargo JA
影响因子：
10.9
作者：
Giraldo NA;Nguyen P;Engle EL;Kaunitz GJ;Cottrell TR;Berry S;Green B;Soni A;Cuda JD;Stein JE;Sunshine JC;Succaria F;Xu H;Ogurtsova A;Danilova L;Church CD;Miller NJ;Fling S;Lundgren L;Ramchurren N;Yearley JH;Lipson EJ;Cheever M;Anders RA;Nghiem PT;Topalian SL;Taube JM
通讯作者：
Taube JM
DOI：
10.1056/nejmoa1504030
发表时间：
2015-07-02
期刊：
The New England journal of medicine
影响因子：
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作者：
Larkin J;Chiarion-Sileni V;Gonzalez R;Grob JJ;Cowey CL;Lao CD;Schadendorf D;Dummer R;Smylie M;Rutkowski P;Ferrucci PF;Hill A;Wagstaff J;Carlino MS;Haanen JB;Maio M;Marquez-Rodas I;McArthur GA;Ascierto PA;Long GV;Callahan MK;Postow MA;Grossmann K;Sznol M;Dreno B;Bastholt L;Yang A;Rollin LM;Horak C;Hodi FS;Wolchok JD
通讯作者：
Wolchok JD
影响因子：
5
作者：
通讯作者：
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