Advances in Multispectral Live-Cell Imaging
Discover how Chroma’s filters are advancing multispectral live-cell imaging!
A research paper, entitled “Multispectral live-cell imaging with uncompromised spatiotemporal resolution,” describes an innovative method for accurate unmixing of low signal-to-noise datasets, enabling detailed cellular studies.
The Abstract of this paper states:
Multispectral imaging is an established method to extend the number of colours usable in fluorescence imaging beyond the typical limit of three or four, but standard approaches are poorly suited to live-cell imaging. We introduce an approach for multispectral imaging in live cells, comprising an iterative spectral unmixing algorithm and eight channel camera-based image acquisition hardware. This enables the accurate unmixing of low signal-to-noise ratio datasets, typical of live-cell imaging, captured at video rates. We use this approach on a commercial spinning disk confocal microscope and a home-built oblique plane light sheet microscope to image 1–7 spectrally distinct fluorophore species simultaneously, using both fluorescent protein fusions and small-molecule dyes. We further use de novo designed protein-binding proteins (minibinders), labelled with organic fluorophores, and use these in combination with our multispectral imaging approach to study the endosomal trafficking of cell-surface receptors at endogenous levels.
Products from Chroma Technology used in this research include:
- Seven dichroic mirrors to span the visible spectrum in roughly equal steps:
- zt532rdc
- zt594rdc
- zt670rdc-xxrxt
- zt491rdc
- zt458rdc
- zt514rdc
- zt561rdc
- ZET405/470/555/640rpc-UF1 quad-band dichroic mirror
- 525/50 for EGFP single-bandpass filter
About Chroma Technology
Chroma Technology is a leading manufacturer and OEM supplier of high performance optical filters used in a wide range of applications, including health care, biomedical research, remote sensing, machine vision, and aerospace. Chroma offers an extensive selection of optical filter sets and filter accessories, as well as custom solutions tailored to customer-specific needs.
For more information or a quotation for this or other Chroma products, contact IL Photonics.