TUK Solution for multicolor, HB Solution
Correlative light-electron microscopy (CLEM) is designed to observe the same sample under both fluorescence microscopy and electron microscopes, allowing comparison of the respective images in order to analyze the localization and morphology of organelles and molecules in cells. Conventionally, the fluorescence of fluorescent proteins was attenuated by osmium oxide used to prepare electron microscopy samples, resulting in chemical/physical strain, which made it difficult to observe the same section under both fluorescence microscopy and electron microscopes.
Fluorescence recovery reagents for CLEM (TUK Solution) can be used together with specific fluorescent proteins (CLEM-Red and CLEM-Green) to recover osmium oxide-quenched fluorescence in samples. As a result, the same section can be observed under both fluorescence microscopy and electron microscopes, leading to more precise CLEM.
Difference between Conventional CLEM and Precision CLEM
Application
Observation of mitochondria
Data by courtesy of Dr. Uchiyama and Dr. Tanida, Graduate School of Medicine, Juntendo University
Figure 1. Mitochondria with recovered fluorescence in CLEM-Red expressing HeLa cells.
Observation of nucleus and mitochondria
Data by I. Tanida.et al. : Scientific Reports ,10,21871 (2020) / Licenced under CC BY 4,0
Figure 2. Nucleus and mitochondria with recovered fluorescence in CLEM-Green and CLEM-Red expressing HEK293 cells.
Immuno-in-resin CLEM
It has been reported that immunological and affinity-labeling using fluorescent dyes applied to in-resin CLEM of Epon-embedded cells.
HB Solution is used for antibody dilution solution and blocking solution.
Data by I. Tanida.et al. : Heliyon, 9(6), e17394 (2023) / Licenced under CC BY 4,0
Figure 3. Two-color in-resin CLEM of intracellular organelles in Epon-embedded cells was performed using Alexa Fluor 647-conjugated WGA and rabbit polyclonal anti-TOM20.
Anti-TOM20 was labeled with SPICARed-streptavidin after biotin labeling.
References
- Tanida, I., Furuta, Y., Yamaguchi, J., Kakuta, S., Oliva Trejo, J. A. and Uchiyama, Y. : Sci. Rep., 10(1), 21871 (2020).
- Sanada, T., Yamaguchi, J., Furuta, Y., Kakuta, S., Tanida, I. and Uchiyama, Y. : Sci. Rep., 12, 11130 (2022).
- Tanida, I., Yamaguchi, J., Suzuki, C., Kakuta, S. and Uchiyama, Y. : Heliyon, . 9(6), e17394, (2023).
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Fluorescence recovery reagent for CLEM
Wash reagent for CLEM
For research use or further manufacturing use only. Not for use in diagnostic procedures.
Product content may differ from the actual image due to minor specification changes etc.
If the revision of product standards and packaging standards has been made, there is a case where the actual product specifications and images are different.