Abstract
Imaging mass cytometry (IMC) is a technique allowing visualization and quantification of over 40 biological parameters in a single experiment with subcellular spatial resolution, however most IMC experiments are limited to endpoint analysis with antibodies and DNA stains. Small molecules containing tellurium are promising probes for IMC due to their cell permeability, synthetic versatility, and most importantly their application to sequential labelling with isotopologous probes (SLIP) experiments. SLIP experiments with tellurium-containing probes allow quantification of intracellular biology at multiple timepoints with IMC. Despite the promise of tellurium in IMC, there are unique challenges in image processing associated with tellurium IMC data. Here, we address some of these issues by demonstrating the removal of xenon background signal, combining channels to improve signal-to-noise ratio, and calculating isotope transmission efficiency biases. These developments add accuracy to the unique temporal resolution afforded by tellurium IMC probes.
Highlights
The emerging technology of imaging mass cytometry (IMC) has delivered insight into many aspects of biology including the heterogeneity of breast cancer tumours and the tissue distribution of cisplatin. [1, 2] Tissue sections are stained with more than 40 different antibodies, each of which is conjugated to a polymer that chelates a distinct elemental isotope, typically from the lanthanide series
Given the specific benefits of tellurophene probes, we sought to design data processing strategies that would improve the accuracy of research in this field and allow sequential labelling with isotopologous probes (SLIP) experiments to be included as robust methods in the IMC community
We have described strategies that enhance the analysis of IMC experiments that use tellurium probes
Summary
The emerging technology of imaging mass cytometry (IMC) has delivered insight into many aspects of biology including the heterogeneity of breast cancer tumours and the tissue distribution of cisplatin. [1, 2] Tissue sections are stained with more than 40 different antibodies, each of which is conjugated to a polymer that chelates a distinct elemental isotope, typically from the lanthanide series. A multi-channel image is thereby created with many more channels accessible than traditional immunohistochemical or immunofluorescent optical imaging This technique has been limited to the imaging of static biological markers (i.e. proteins), DNA, or certain small molecule probes against which custom antibodies have been raised (e.g. EF5). [4] Probes for specific biological processes can be synthesized by linking tellurophenes, which are aromatic, stable, and non-toxic, to activitybased functional groups that covalently bind cells of a certain phenotype.
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