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Filter-Dense Multicolor Microscopy

Artikel i vetenskaplig tidskrift
Författare Siavash Kijani
Ulf Yrlid
Maria Heyden
Malin Levin
Jan Borén
Per Fogelstrand
Publicerad i Plos One
Volym 10
Nummer/häfte 3
ISSN 1932-6203
Publiceringsår 2015
Publicerad vid Wallenberglaboratoriet
Institutionen för biomedicin, avdelningen för mikrobiologi och immunologi
Institutionen för medicin, avdelningen för molekylär och klinisk medicin
Språk en
Länkar dx.doi.org/10.1371/journal.pone.011...
Ämnesord CELL SUBSETS, ATHEROSCLEROSIS, FLUORESCENCE, IMMUNOFLUORESCENCE, MACROPHAGES, DISTINCT, Multidisciplinary Sciences
Ämneskategorier Immunologi inom det medicinska området

Sammanfattning

Immunofluorescence microscopy is a unique method to reveal the spatial location of proteins in tissues and cells. By combining antibodies that are labeled with different fluorochromes, the location of several proteins can simultaneously be visualized in one sample. However, because of the risk of bleed-through signals between fluorochromes, standard multicolor microscopy is restricted to a maximum of four fluorescence channels, including one for nuclei staining. This is not always enough to address common scientific questions. In particular, the use of a rapidly increasing number of marker proteins to classify functionally distinct cell populations and diseased tissues emphasizes the need for more complex multistainings. Hence, multicolor microscopy should ideally offer more channels to meet the current needs in biomedical science. Here we present an enhanced multi-fluorescence setup, which we call Filter-Dense Multicolor Microscopy (FDMM). FDMM is based on condensed filter sets that are more specific for each fluorochrome and allow a more economic use of the light spectrum. FDMM allows at least six independent fluorescence channels and can be applied to any standard fluorescence microscope without changing any operative procedures for the user. In the present study, we demonstrate an FDMM setup of six channels that includes the most commonly used fluorochromes for histology. We show that the FDMM setup is specific and robust, and we apply the technique on typical biological questions that require more than four fluorescence microscope channels.

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