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Chemistry

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Methods Article

Porphyrin-Modified Beads for Use as Compensation Controls in Flow Cytometry

2.0K Views

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10:06 min

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March 24th, 2023

DOI :

10.3791/65294-v

March 24th, 2023

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William Bauta¹, Marcia Grayson¹, Rossella Titone¹, Jennifer Rebeles¹, Vivienne I. Rebel¹

¹bioAffinity Technologies

Transcript

We developed polystyrene beads modified with the porphyrin TCPP as compensation controls and multicolor flow cytometry. The beads used in this protocol have fluorescence intensity comparable to TCPP stain cells and still passed through the flow cytometer. We accomplished that and are currently working on better methods to remove the particulate byproduct.

Porphyrin-modified compensation beads were preferable to TCPP stained cells as compensation controls. These speeds were stable over months without significant loss of fluorescence. Porphyrins are known to stain cancer cells selectively.

The research combines the selectivity of porphyrins with quantitative flow cytometry as a powerful platform for high-throughput analysis of human samples.

Summary

The protocol describes how porphyrin-based compensation beads for flow cytometry are prepared by the reaction of amine-functionalized polystyrene beads with the porphyrin TCPP and the amide coupling reagent EDC. A filtration procedure is used to reduce the particulate byproducts.

Explore More Videos

Porphyrin modified Beads

Compensation Controls

Polystyrene Beads

Fluorescence Intensity

Cancer Cell Staining

Quantitative Analysis

Fluorescent Reagents

Fluorescence Measurements

Compensation Process

Amine functionalized Beads

EDC Coupling Reagent

High throughput Analysis

Chapters in this video

0:00

Introduction

0:53

Preparation of Meso‐Tetra(4‐Carboxyphenyl) Porphine (TCPP) Stock Solution

2:20

Combining Amine‐Functionalized Polystyrene Beads with TCPP Solution

3:15

Labeling Amine‐Functionalized Polystyrene Beads with TCPP

5:21

Quality Check (QC) of the TCPP Beads by Flow Cytometry

7:13

TCPP Bead Filtration

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