Fluorescence in situ hybridization assays were performed using the RiboTM Fluorescent in situ Hybridization Kit and the RiboTM lncRNA FISH Probe Mix (Ribo, Guangzhou, China) according to the manufacturer’s protocols. Briefly, the cells were fixed in 4% formaldehyde for 10 min and then washed with PBS. The cells were incubated with 20 μmol/L lncRNA FISH probe mix at 37°C overnight. After washing, the FISH preparations were counterstained with DAPI observed in confocal microscopy for appropriate fluorescence filter sets (Zeiss, Oberkochen, Germany). The lncRNA probe labeled with Cy3 was designed and synthesized by RiboBio Co., Ltd., RiboTM U6 and RiboTM 18S were used as reference controls for the subcellular localization of lncRNA.
Fluorescence filter sets
Manufactured by Zeiss
Sourced in Germany
Fluorescence filter sets are optical components used in fluorescence microscopy to selectively transmit specific wavelengths of light. They consist of excitation, emission, and dichroic filters that isolate the desired fluorescence signal from the excitation light. These filter sets are designed to work with various fluorescent dyes and proteins to enable the visualization and analysis of biological samples.
Automatically generated - may contain errors
2 protocols using fluorescence filter sets
1
Fluorescent In Situ Hybridization for lncRNA
2
Multimodal Microscopy Imaging Protocol
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Immunofluorescence images were taken with an inverted Zeiss Axio Observer microscope equipped with an ApoTome.2, an Axiocam 702 mono and Colibri 7 illumination system. In addition an inverted Zeiss Axio Imager microscope was used. 100x, 63x, 40x, 20x and 10x objectives with Zeiss fluorescence filter sets (49 DAPI, 38 GFP, 43 HE dsRed, 50 Cy5) were used for immunofluorescence and 40x and 20x objectives for phase-contrast analysis.
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