20.500.12556/RUL-184867 Correlative light and electron microscopy in cell biology accessible pre-embedding and post-embedding strategies Correlative light and electron microscopy (CLEM) combines the molecular specificity of fluorescence light microscopy with the ultrastructural resolution of electron microscopy (EM), enabling molecularly defined cellular structures to be localised and interpreted within their cellular ultrastructural context. Although cryo-CLEM, super-resolution CLEM, and volume EM have substantially expanded the spatial, molecular, and three-dimensional capabilities of CLEM, their routine implementation remains constrained by specialised instrumentation, demanding sample-preparation workflows, and the need for expertise in image acquisition, registration, and analysis. This review therefore focuses on conventional CLEM strategies that are accessible to laboratories equipped with standard widefield or confocal fluorescence microscopes and transmission electron microscopes. Pre-embedding and post-embedding approaches are compared, highlighting how the timing of labelling and fluorescence imaging influences key trade-offs among live-cell imaging, correlation accuracy, probe preservation, and ultrastructural integrity. Probes used in pre-embedding workflows are further evaluated, including dual-modality probes such as FluoroNanogold antibodies and quantum dots, fluorescent proteins like GFP (rendered EM-visible through secondary immunolabelling or photoconversion), and luminescent metal complexes, alongside selected protocol examples for both pre-embedding and post-embedding experimental approaches. Overall, successful CLEM depends on careful experimental design, appropriate probe selection, and optimised sample preparation. Conventional workflows remain practical and robust options for routine cell biology studies, particularly when accessibility, reproducibility, and compatibility with existing microscopy infrastructure are priorities. correlative light and electron microscopy pre-embedding post-embedding correlative imaging multimodal probes sample preparation korelativna svetlobna in elektronska mikroskopija pred vklopitvijo po vklopitvi korelativno slikanje multimodalne sonde priprava vzorcev true false true Angleški jezik Slovenski jezik Članek v reviji 2026-07-16 09:47:50 2026-07-16 09:48:01 2026-07-17 04:30:51 0000-00-00 00:00:00 2026 0 0 Str. 141-154 no. 2 Vol. 45 Jun. 2026 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 57.086 1580-3139 10.5566/ias.4011 283977219 RAZ_Janev_Aleksandar_2026.pdf RAZ_Janev_Aleksandar_2026.pdf 1 8DB3F16146DB4C4D6062E291B99E946D b35c1a691c57ad50f21ba22dc9a5794acb2caef8324f6c7fe24b52002c7cc7cb bc0b9b74-80ea-11f1-9b0d-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=240240 https://www.ias-iss.org/ojs/IAS/article/view/4011 1 7da8df37-80ea-11f1-9b0d-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=240239 Medicinska fakulteta 0 0 0