Summary of 300-Nishiyama-Rdot

SSBD:database
SSBD:database URL
Title
MCF-7 cells stained by 12 different colors of Rdots
Description
-
Relase date
2024-11-25
Updated date
-
License
CC BY
Kind
Image data based on Experiment
Number of Datasets
1 ( Image datasets: 1, Quantitative data datasets: 0 )
Size of Datasets
635.4 MB ( Image datasets: 635.4 MB, Quantitative data datasets: 0 bytes )

Organism(s)
Homo sapiens
Cell lines(s)
MCF7 cell
Gene symbol(s)
NA
Protein name(s)
NA

Datatype
-
Molecular Function (MF)
Biological Process (BP)
Cellular Component (CC)
Biological Imaging Method
inelastic scattering of photons
T scale
-

Image Acquisition
Experiment type
-
Microscope type
-
Acquisition mode
-
Contrast method
-
Microscope model
-
Detector model
-
Objective model
-
Filter set
-

Related paper(s)

Ryo Nishiyama, Kotaro Hiramatsu, Shintaro Kawamura, Kosuke Dodo, Kei Furuya, Julia Gala de Pablo, Shigekazu Takizawa, Wei Min, Mikiko Sodeoka, Keisuke Goda (2023) Color-scalable flow cytometry with Raman tags., PNAS Nexus, Volume 2, Number 2

Published in 2023 Feb (Electronic publication in Jan. 14, 2023, midnight )

(Abstract) Flow cytometry is an indispensable tool in biology and medicine for counting and analyzing cells in large heterogeneous populations. It identifies multiple characteristics of every single cell, typically via fluorescent probes that specifically bind to target molecules on the cell surface or within the cell. However, flow cytometry has a critical limitation: the color barrier. The number of chemical traits that can be simultaneously resolved is typically limited to several due to the spectral overlap between fluorescence signals from different fluorescent probes. Here, we present color-scalable flow cytometry based on coherent Raman flow cytometry with Raman tags to break the color barrier. This is made possible by combining a broadband Fourier-transform coherent anti-Stokes Raman scattering (FT-CARS) flow cytometer, resonance-enhanced cyanine-based Raman tags, and Raman-active dots (Rdots). Specifically, we synthesized 20 cyanine-based Raman tags whose Raman spectra are linearly independent in the fingerprint region (400 to 1,600 cm−1). For highly sensitive detection, we produced Rdots composed of 12 different Raman tags in polymer nanoparticles whose detection limit was as low as 12 nM for a short FT-CARS signal integration time of 420 µs. We performed multiplex flow cytometry of MCF-7 breast cancer cells stained by 12 different Rdots with a high classification accuracy of 98%. Moreover, we demonstrated a large-scale time-course analysis of endocytosis via the multiplex Raman flow cytometer. Our method can theoretically achieve flow cytometry of live cells with >140 colors based on a single excitation laser and a single detector without increasing instrument size, cost, or complexity.

Contact
Kotaro Hiramatsu, Keisuke Goda , Kyusyu University, The University of Tokyo , Faculty of Sciences Department of Chemistry, Department of Chemistry
Contributors
Ryo Nishiyama, Shintaro Kawamura, Kosuke Dodo, Mikiko Sodeoka


Dataset List of 300-Nishiyama-Rdot

#
Dataset ID
Kind
Size
4D View
SSBD:OMERO
Download BDML
Download Images
# 11577
Datast ID Fig2C_Rdots
Dataset Kind Image data
Dataset Size 635.4 MB
4D view
SSBD:OMERO
Download BDML
Download Image data