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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.
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.
Name | Company | Catalog Number | Comments |
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 |
This article has been published
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Source: Yoon, J. et al. Label-Free Identification of Lymphocyte Subtypes Using Three-Dimensional Quantitative Phase Imaging and Machine Learning. J. Vis. Exp. (2018)
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