scholarly journals Sensitive detection of RNAs in single cells by flow cytometry

1992 ◽  
Vol 20 (1) ◽  
pp. 83-88 ◽  
Author(s):  
Hong Yu ◽  
Lauren Ernst ◽  
Marc Wagner ◽  
Alan Waggoner
Keyword(s):  
Flow Cytometry ◽  
Single Cells ◽  
Sensitive Detection

scholarly journals Sensitive detection of RNAs in single cells by flow cytometry

1992 ◽  
Vol 20 (20) ◽  
pp. 5518-5518
Author(s):  
H. Yu ◽  
L. Ernst ◽  
M. Wagner ◽  
A. Waggoner
Keyword(s):  
Flow Cytometry ◽  
Single Cells ◽  
Sensitive Detection

Highly sensitive detection of multiple proteins from single cells by MoS2-FET biosensors

Talanta ◽  
2021 ◽  
pp. 122839
Author(s):  
Junqing Wei ◽  
Zhihan Zhao ◽  
Kuibo Lan ◽  
Zhi Wang ◽  
Guoxuan Qin ◽  
...  
Keyword(s):  
Single Cells ◽  
Sensitive Detection ◽  
Highly Sensitive ◽  
Highly Sensitive Detection

scholarly journals A selective cytotoxic adenovirus vector for concentration of pluripotent stem cells in human pluripotent stem cell-derived neural progenitor cells

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Takamasa Hirai ◽  
Ken Kono ◽  
Rumi Sawada ◽  
Takuya Kuroda ◽  
Satoshi Yasuda ◽  
...  
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Stem Cell ◽  
Progenitor Cells ◽  
Pluripotent Stem Cells ◽  
Pluripotent Stem Cell ◽  
Neural Progenitor Cells ◽  
Neural Progenitor ◽  
Sensitive Detection ◽  
Quality And Safety

AbstractHighly sensitive detection of residual undifferentiated pluripotent stem cells is essential for the quality and safety of cell-processed therapeutic products derived from human induced pluripotent stem cells (hiPSCs). We previously reported the generation of an adenovirus (Ad) vector and adeno-associated virus vectors that possess a suicide gene, inducible Caspase 9 (iCasp9), which makes it possible to sensitively detect undifferentiated hiPSCs in cultures of hiPSC-derived cardiomyocytes. In this study, we investigated whether these vectors also allow for detection of undifferentiated hiPSCs in preparations of hiPSC-derived neural progenitor cells (hiPSC-NPCs), which have been expected to treat neurological disorders. To detect undifferentiated hiPSCs, the expression of pluripotent stem cell markers was determined by immunostaining and flow cytometry. Using immortalized NPCs as a model, the Ad vector was identified to be the most efficient among the vectors tested in detecting undifferentiated hiPSCs. Moreover, we found that the Ad vector killed most hiPSC-NPCs in an iCasp9-dependent manner, enabling flow cytometry to detect undifferentiated hiPSCs intermingled at a lower concentration (0.002%) than reported previously (0.1%). These data indicate that the Ad vector selectively eliminates hiPSC-NPCs, thus allowing for sensitive detection of hiPSCs. This cytotoxic viral vector could contribute to ensuring the quality and safety of hiPSCs-NPCs for therapeutic use.

Fast intracellular pH determination in single cells by flow-cytometry

1981 ◽  
Vol 68 (5) ◽  
pp. 265-266 ◽  
Author(s):  
G. Valet ◽  
A. Raffael ◽  
L. Moroder ◽  
E. W�nsch ◽  
G. Ruhenstroth-Bauer
Keyword(s):  
Flow Cytometry ◽  
Intracellular Ph ◽  
Single Cells

scholarly journals Glyoxal fixation facilitates transcriptome analysis after antigen staining and cell sorting by flow cytometry

PLoS ONE ◽  
2021 ◽  
Vol 16 (1) ◽  
pp. e0240769
Author(s):  
Prasanna Channathodiyil ◽  
Jonathan Houseley
Keyword(s):  
Flow Cytometry ◽  
Transcriptome Analysis ◽  
Mammalian Cells ◽  
Single Cells ◽  
Rna Extraction ◽  
Cyclin B1 ◽  
Immunofluorescent Staining ◽  
Rna Seq ◽  
Simple Method ◽  
Staining Methods

A simple method for extraction of high quality RNA from cells that have been fixed, stained and sorted by flow cytometry would allow routine transcriptome analysis of highly purified cell populations and single cells. However, formaldehyde fixation impairs RNA extraction and inhibits RNA amplification. Here we show that good quality RNA can be readily extracted from stained and sorted mammalian cells if formaldehyde is replaced by glyoxal—a well-characterised fixative that is widely compatible with immunofluorescent staining methods. Although both formaldehyde and glyoxal efficiently form protein-protein crosslinks, glyoxal does not crosslink RNA to proteins nor form stable RNA adducts, ensuring that RNA remains accessible and amenable to enzymatic manipulation after glyoxal fixation. We find that RNA integrity is maintained through glyoxal fixation, permeabilisation with methanol or saponin, indirect immunofluorescent staining and flow sorting. RNA can then be extracted by standard methods and processed into RNA-seq libraries using commercial kits; mRNA abundances measured by poly(A)+ RNA-seq correlate well between freshly harvested cells and fixed, stained and sorted cells. We validate the applicability of this approach to flow cytometry by staining MCF-7 cells for the intracellular G2/M-specific antigen cyclin B1 (CCNB1), and show strong enrichment for G2/M-phase cells based on transcriptomic data. Switching to glyoxal fixation with RNA-compatible staining methods requires only minor adjustments of most existing staining and sorting protocols, and should facilitate routine transcriptomic analysis of sorted cells.

scholarly journals Flow Cytometric Assessment of Susceptibilities of Streptococcus pyogenes to Erythromycin and Rokitamycin

2003 ◽  
Vol 47 (1) ◽  
pp. 408-412 ◽  
Author(s):  
Pier Carlo Braga ◽  
Cinzia Bovio ◽  
Maria Culici ◽  
Monica Dal Sasso
Keyword(s):  
Flow Cytometry ◽  
Fluorescence Microscopy ◽  
Streptococcus Pyogenes ◽  
Single Cells ◽  
Membered Ring ◽  
Flow Cytometric ◽  
Live Bacteria ◽  
Erythromycin A ◽  
Set Up ◽  
Syto 9

ABSTRACT The effects of erythromycin (a 14-membered ring macrolide) and rokitamycin (a 16-membered ring macrolide) on the viability of the Streptococcus pyogenes M phenotype were studied by means of flow cytometry and fluorescence microscopy by using a combination of two fluorochromes (syto 9 and propidium iodide) that stains live bacteria green and dead bacteria red. In order to apply the flow cytometry, a bacterial sonication procedure was expressly set up to separate single cells from the long, intralaced S. pyogenes chains of up to 30 to 40 cells that have previously prevented the application of flow cytometry to this type of bacteria. The association of flow cytometry using an appropriate sonication procedure, together with a combination of fluorescent probes, offered the possibility of very quickly investigating the different microbiological effects of rokitamycin at 2 μg/ml, which was active on the S. pyogenes M phenotype, and of erythromycin at doses of up to 32 μg/ml, which was not.

scholarly journals Estimating RNA numbers in single cells by RNA fluorescent tagging and flow cytometry

2019 ◽  
Vol 166 ◽  
pp. 105745 ◽  
Author(s):  
Mohamed N.M. Bahrudeen ◽  
Vatsala Chauhan ◽  
Cristina S.D. Palma ◽  
Samuel M.D. Oliveira ◽  
Vinodh K. Kandavalli ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Single Cells ◽  
Fluorescent Tagging

Flow cytometry: a powerful tool in analysis of biomass distributions in phytoplankton

1995 ◽  
Vol 32 (4) ◽  
pp. 177-182 ◽  
Author(s):  
R. R. Jonker ◽  
J. T. Meulemans ◽  
G. B. J. Dubelaar ◽  
M. F. Wilkins ◽  
J. Ringelberg
Keyword(s):  
Flow Cytometry ◽  
Large Range ◽  
Single Cells ◽  
Cell Types ◽  
Spectrometric Analysis ◽  
Analysis Techniques ◽  
Field Samples ◽  
Phytoplankton Monitoring ◽  
Correlated Information ◽  
Taxonomic Groups

The large range in concentrations and cell-sizes of algal cells and colonies and the large variety of cell types are the main reasons for developing a dedicated cytometer for the analysis of phytoplankton. A European Community funded consortium has developed the EurOPA cytometer, which is easily transported and can be operated at sea. With the EurOPA, both small single cells and large colonies of cyanobacteria can be analyzed in one run. This provides correlated information on optical characteristics, pigments contents and taxonomy. The resulting distribution of (chlorophyll) biomass over taxonomic groups can be inter-calibrated with standard spectrometric analysis techniques. The EurOPA can be used successfully for analysis of field samples and phytoplankton cultures. It is well suited for phytoplankton monitoring and grazing studies.

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