Palladium Barcoding
Palladium Barcoding - Related Videos
06:19
Palladium-Catalyzed Cross Coupling
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2023
Source: Vy M. Dong and Faben Cruz, Department of Chemistry, University of California, Irvine, CA This experiment will demonstrate the concept of a palladium-catalyzed cross coupling. The set-up of a typical Pd-catalyzed cross coupling reaction will be illustrated. Pd-catalyzed cross coupling reactions have had a profound effect on how synthetic chemists create molecules. These reactions have enabled chemists to construct bonds in new and more efficient ways. Such reactions have found widespread...
05:42
A Barcoding Assay for Phenotypic and Functional Characterization of Immune Cells
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2025
This video highlights a single-cell proteomic method that employs heavy-metal-conjugated antibodies for distinct cell barcoding. Subsequently, immunostaining and mass cytometry are applied to evaluate immune phenotypic and functional alterations in human whole blood-derived immune cells.
13:14
Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications
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Cited by 4 •
2015
Since the discovery of the green fluorescent protein gene, fluorescent proteins have impacted molecular cell biology. This protocol describes how expression of distinct fluorescent proteins through genetic engineering is used for barcoding individual cells. The procedure enables tracking distinct populations in a cell mixture, which is ideal for multiplexed applications.
11:59
Competitive Genomic Screens of Barcoded Yeast Libraries
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Cited by 44 •
2011
We have developed comprehensive, unbiased genome-wide screens to understand gene-drug and gene-environment interactions. Methods for screening these mutant collections are presented.
12:59
BEST: Barcode Enabled Sequencing of Tetrads
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Cited by 4 •
2014
Barcode Enabled Sequencing of Tetrads (BEST) replaces the manual processes of isolating, disrupting and spacing tetrads. BEST isolates tetrads by fluorescence-activated cell sorting onto agar plates, separates the spores by agitation with glass beads, and determines which randomly arrayed colonies were derived from the same original tetrad using molecular barcodes.
FAQs
Each sample receives a distinct combination of palladium isotope tags rather than relying on a single label. After the barcoded samples are pooled, time-of-flight mass spectrometry measures their isotope patterns and assigns events back to the appropriate sample. This decoding step preserves sample identity while allowing pooled material to undergo downstream high-dimensional analysis in one tube.
Fixation and permeabilization create the conditions needed for the reagents to enter cells and react with intracellular proteins. The reactive palladium complexes bind those proteins, producing an isotope signature associated with the corresponding sample during pooled analysis. This chemistry is important because the workflow encodes sample identity inside cellular material rather than depending only on separate sample handling.
Pooling barcoded conditions in one tube makes them undergo the same subsequent sample handling and instrument measurement. That arrangement supports direct comparisons while reducing staining and instrument variability between separately processed conditions. It also increases throughput because several conditions can be analyzed together, making the approach useful when experiments require broad, high-dimensional immune profiling.
Once sample identities can be recovered from pooled data, antibody measurements from many conditions can be examined together without losing the condition associated with each cell. High-dimensional staining then supports separation of diverse immune-cell phenotypes and detection of changes within complex populations. This is particularly valuable when a response is confined to a rare subset rather than the overall cell mixture.
Cells or samples are first assigned distinct palladium isotope combinations, then fixed and permeabilized so the reagents can enter and bind intracellular proteins. Barcoded material from the conditions is pooled into one tube. The pooled sample can undergo antibody staining and is subsequently measured by time-of-flight mass spectrometry, which uses isotope patterns to recover sample identity.
The approach can connect immune-cell phenotype with signaling responses and pathogen-induced changes within the same high-dimensional experiment. Antibody staining supplies the measurements used to characterize immune cells, while time-of-flight mass spectrometry records the isotope information needed to assign observations to their original conditions. The result is a multiplexed view of how different samples or treatments affect immune populations.
It is useful when many experimental conditions must be compared directly and researchers want to limit variability introduced by separate staining or instrument runs. By combining conditions in one tube, the method improves throughput and supports coordinated high-dimensional measurements. This makes it relevant for experiments examining multiple immune states or infection-related responses.
Immune responses to infection can vary across cell types and conditions, so retaining each sample’s identity during pooled, high-dimensional measurement is important. Palladium barcoding allows phenotypes, signaling responses, and pathogen-induced changes to be compared across those conditions. Its ability to reveal heterogeneous or rare populations adds resolution when infection-associated effects are not uniform across immune cells.