Methods for analyzing tellurium imaging mass cytometry data

Bassan, Jay; Nitz, Mark

doi:10.1371/journal.pone.0221714

Cited by 6 publications

(6 citation statements)

References 13 publications

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“…Images were received as .txt files from the instrument. Each image was unpacked into separate numpy arrays for each mass channel using teimc package [54] . Numpy, [55] pandas, [56] Matplotlib, [53] and scikit‐image [57] libraries were then used for analysis of raw images.…”

Section: Methodsmentioning

confidence: 99%

An Iodinated DAPI‐Based Reagent for Mass Cytometry

et al. 2020

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Multiparametric single‐cell analysis has seen dramatic improvements with the introduction of mass cytometry (MC) and imaging mass cytometry (IMC™). These technologies expanded the number of biomarkers that can be identified simultaneously by using heavy‐isotope‐tagged antibody reagents. Small‐molecule probes bearing heavy isotopes are emerging as additional useful functional reporters of cellular features. Realizing this, we explored the iodination of DAPI to produce a heavy‐atom‐substituted derivative of the commonly used fluorescent DNA stain. Although exhibiting a drastically reduced fluorescence emission profile, I‐DAPI retains strong binding affinity for DNA. I‐DAPI was used to detect cellular DNA in MC and IMC™ assays with comparable efficiency to known Ir‐containing DNA intercalators. This work suggests repurposing well‐known colorimetric stains through simple reactions could be an effective strategy to develop new, functional MC and IMC™ reagents.

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Section: Methodsmentioning

confidence: 99%

An Iodinated DAPI‐Based Reagent for Mass Cytometry

et al. 2020

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“…Figure 28 shows an IMC image of TePhe incorporation into a mouse jejunum stained with a DNA intercalator and Maxpar-labeled antialpha smooth muscle actin, with a stronger Te signal observed in the crypts than the villi. Bassan and Nitz 410 further optimized the analysis of Te probes by developing protocols for removing overlap with the Te signal and combining Te isotopic signals to improve signal-to-noise. Here, the authors assembled a three-dimensional stack of all isotopic images of Te and applied a Gaussian blur filter to reduce noise.…”

Section: Imaging Mass Cytometrymentioning

confidence: 99%

Laser Ablation–Inductively Coupled Plasma–Mass Spectrometry Imaging in Biology

et al. 2021

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Elemental imaging gives insight into the fundamental chemical makeup of living organisms. Every cell on Earth is comprised of a complex and dynamic mixture of the chemical elements that define structure and function. Many disease states feature a disturbance in elemental homeostasis, and understanding how, and most importantly where, has driven the development of laser ablation−inductively coupled plasma−mass spectrometry (LA-ICP-MS) as the principal elemental imaging technique for biologists. This review provides an outline of ICP-MS technology, laser ablation cell designs, imaging workflows, and methods of quantification. Detailed examples of imaging applications including analyses of cancers, elemental uptake and accumulation, plant bioimaging, nanomaterials in the environment, and exposure science and neuroscience are presented and discussed. Recent incorporation of immunohistochemical workflows for imaging biomolecules, complementary and multimodal imaging techniques, and image processing methods is also reviewed.CONTENTS

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“…Efforts to combine intensities from multiple tellurium channels to improve the signal-to-noise ratio were also unsuccessful. [25] We hypothesized that the processing of the FFPE tissues contributed to loss of tellurium signal as the teniposide interaction with the cells is largely reversible, thus the multiple wash steps required for dewaxing (xylene and ethanol washes) might remove bound 1. Initial attempts using the frozen tissue IMC™ protocol (FLDM-00073) involving fixing with 4 % PFA and washing were also unsuccessful -tellurium signal in xenograft sections from mice treated with 1 was indistinguishable from untreated tissue samples.…”

Section: In Vivo Evaluation Of Tellurium-labelled Teniposidementioning

confidence: 99%

Tellurophene‐Tagging of Teniposide Facilitates Monitoring by Mass Cytometry

et al. 2022

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Target engagement and the biodistribution of exogenously administered small molecules is rarely homogenous. Methods to determine the biodistribution at the cellular level are limited by the ability to detect the small molecule and simultaneously identify the cell types or tissue structures with which it is associated. The highly multiplexed nature of mass cytometry could facilitate these studies provided a heavy isotope label was available in the molecule of interest. Here we show it is possible to append a tellurophene to a known chemotherapeutic, teniposide, to follow this molecule in vivo. A semi-synthetic approach offers an efficient route to the teniposide analogue which is found to have similar characteristics when compared with the parent teniposide in vitro. Using mass cytometry we find the teniposide analogue has significant nonspecific binding to cells. In vivo the tellurium bearing teniposide produces the expected DNA damage in a PANC-1 xenograft model. The distribution of Te in the tissue is near the limits of detection and further work will be required to characterize the localization of this analogue with respect to cell type distributions.

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Methods for analyzing tellurium imaging mass cytometry data

Cited by 6 publications

References 13 publications

An Iodinated DAPI‐Based Reagent for Mass Cytometry

An Iodinated DAPI‐Based Reagent for Mass Cytometry

Laser Ablation–Inductively Coupled Plasma–Mass Spectrometry Imaging in Biology

Tellurophene‐Tagging of Teniposide Facilitates Monitoring by Mass Cytometry

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