Three-Dimensional Quantitative Phase Imaging for Characterizing Lymphocyte Subtypes

Published: November 30, 2023

Abstract

Source: Yoon, J. et al. Label-Free Identification of Lymphocyte Subtypes Using Three-Dimensional Quantitative Phase Imaging and Machine Learning. J. Vis. Exp.  (2018)

This video demonstrates label-free identification of lymphocyte morphology and biochemistry through 3D quantitative phase imaging. This technique allows for a thorough analysis of lymphocytes, uncovering their internal structures and properties without the need for labeling procedures.

Protocol

1. Flow Cytometry and Sorting of Lymphocyte Subtypes NOTE: Sorting lymphocytes depending on cell type is essential for establishing the ground-truth (i.e., correct) cell type labels to train and test a cell type classifier in supervised learning. Flow cytometry, a gold standard method, is used to identify and separate lymphocytes. Make a mixture of surface marker staining antibodies in 100 µL of fresh RPMI-1640 medium [with 10% FBS, 0.1 µg of CD3…

開示

The authors have nothing to disclose.

Materials

Flow cytometry BD Biosciences Aria II or III
Imaging chamber Tomocube, Inc. TomoDish
Holotomography Tomocube, Inc HT-1H
Holotomography imaging software Tomocube, Inc. TomoStudio
Image-processing software MathWorks Matlab R2017b
Falcon conical centrifuge tube ThermoFisher Scientific 14-959-53A 15 mL
Phosphate-buffered saline Sigma-Aldrich 806544-500ML
Ammonium-chloride-potassium lysing buffer ThermoFisher Scientific A1049201
RPMI-1640 medium Sigma-Aldrich R8758
Fetal bovine serum ThermoFisher Scientific 10438018

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記事を引用
Three-Dimensional Quantitative Phase Imaging for Characterizing Lymphocyte Subtypes. J. Vis. Exp. (Pending Publication), e21841, doi: (2023).

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