Microfluidic single-cell analysis of oxidative stress in Dictyostelium discoideum

The Analyst ◽  
2018 ◽  
Vol 143 (15) ◽  
pp. 3643-3650 ◽  
Author(s):  
Kathy Rodogiannis ◽  
Jessica T. Duong ◽  
Michelle L. Kovarik
Keyword(s):  
Oxidative Stress ◽  
Single Cell ◽  
Dictyostelium Discoideum ◽  
Single Cell Analysis ◽  
Cell Analysis ◽  
Powerful Technique ◽  
Chemical Cytometry ◽  
Unicellular Organisms ◽  
First Time ◽  
Chemical Contents

Microfluidic chemical cytometry is a powerful technique for examining chemical contents of individual cells. Here, it is applied to study unicellular organisms for the first time.

Single‐Cell Analysis by Chemical Cytometry Combined with Fluorescence Microscopy

2004 ◽  
Vol 32 (1) ◽  
pp. 31-41 ◽  
Author(s):  
Kang Hu ◽  
Arash Zarrine‐Afsar ◽  
Hossein Ahmadzadeh ◽  
Sergey N. Krylov
Keyword(s):  
Fluorescence Microscopy ◽  
Single Cell ◽  
Single Cell Analysis ◽  
Cell Analysis ◽  
Chemical Cytometry

scholarly journals Single Cell Analysis Proves the Coexistence of NOTCH1 and TP53 Mutations within the Same Cancer Cells in Patients with Chronic Lymphocytic Leukemia

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 2913-2913
Author(s):  
Barbara Kantorova ◽  
Jitka Malcikova ◽  
Kamila Brazdilova ◽  
Marek Borsky ◽  
Karla Plevova ◽  
...  
Keyword(s):  
Single Cell ◽  
Tp53 Mutation ◽  
Single Cell Analysis ◽  
Direct Sequencing ◽  
Gene Mutations ◽  
Lymphocytic Leukemia ◽  
Cell Analysis ◽  
Tp53 Mutations ◽  
First Time ◽  
Clonal Composition

Abstract Introduction Mutations in NOTCH1 and especially TP53 genes represent potent drivers of chronic lymphocytic leukemia (CLL) progression and chemo-refractoriness. Although a coexistence of these mutations was reported in CLL, a molecular basis of this phenomenon has not been described yet. To clarify this issue, we performed a detailed analysis of CLL patients with parallel NOTCH1 and TP53 mutations including single cancer cell examination. Methods TP53 mutations were determined based on FASAY analysis coupled with direct sequencing. In a collected cohort of 111 TP53 -mutated patients a presence of hot spot c.7544_7545delCT NOTCH1 mutation was assessed using direct gDNA sequencing. In NOTCH1 -TP53 -mutated patients with available material, the mutations' coexistence was tested in single flow-sorted CD19+ cells (cancer cell proportion > 80 %) using multiplex PCR followed by direct sequencing. Results The NOTCH1 mutation was detected in 19/111 (17 %) of the TP53 -mutated patients. Eleven of the NOTCH1-TP53 -mutated patients carried single TP53 mutation; multiple TP53 mutations were detected in 8 of them. Based on gDNA sequencing, the NOTCH1 and TP53 mutation coexistence in the same cancer cells was evident in 4/19 of the NOTCH1-TP53-mutated patients, as at least one of the gene mutations occurred in 100 % of the DNA. In the remaining 15 NOTCH1-TP53 -mutated patients the clonal composition was not possible to assess using sequencing data only and therefore a single cell analysis was performed in 8 of them with available material. Remarkably, irrespective of the mutation proportion, in all of these patients the NOTCH1 mutation was always present together with at least one of the detected TP53 mutations. Considering both the DNA sequencing and single cell analysis data, the 12 patientswith proven NOTCH1-TP53 mutation coexistence might be stratified into three groups with different clonal composition: i) patients with NOTCH1 and single TP53 mutations showing a comparable mutation proportion (n = 3), in which both gene mutations were always detected in the same cells and never occurred separately; ii) patients with either NOTCH1 or TP53 mutation predominance (n = 6), in which the predominant mutation was present separately as well as in combination with the coexisting mutation(s) in individual cells; iii) patients with NOTCH1 and multiple TP53 mutations showing different mutation proportion (n = 3), in which NOTCH1 mutation was present together with one of the detected TP53 mutations in the same cells, while the other TP53 mutations occurred separately. In two of the NOTCH1-TP53 -mutated patients who received intensive chemo-immunotherapy, the consecutive samples were available for single cell analysis. In one of these patients only single TP53 mutation was detected at first time point. In relapse after rituximab-dexamethasone treatment the clone carrying the original TP53 mutation expanded in parallel with another NOTCH1-TP53-mutated clone. Different situation was noticed in the second patient, in which the NOTCH1-TP53-mutated clone detected at first time point diminished after alemtuzumab treatment, while another TP53-mutated-NOTCH1-wild-type clone expanded in relapse. Conclusion We have shown that in NOTCH1-TP53 -mutated patients the mutations often coexist in the same CLL cells. These patients exhibit a considerable clonal heterogeneity that may be further influenced by chemo-immunotherapy. This study was supported by IGA NT/13493 and NT/13519, MUNI/A/1180/2014, CZ.1.05/1.1.00/02.0068. Disclosures Mayer: Janssen: Research Funding.

Chemical Cytometry: Fluorescence-Based Single-Cell Analysis

2008 ◽  
Vol 1 (1) ◽  
pp. 165-190 ◽  
Author(s):  
Daniella Cohen ◽  
Jane A. Dickerson ◽  
Colin D. Whitmore ◽  
Emily H. Turner ◽  
Monica M. Palcic ◽  
...  
Keyword(s):  
Single Cell ◽  
Single Cell Analysis ◽  
Cell Analysis ◽  
Chemical Cytometry

Author response for "Immunocyte single cell analysis of vaccine‐induced narcolepsy"

2020 ◽  
Author(s):  
Alexander Lind ◽  
Falastin Salami ◽  
Anne‐Marie Landtblom ◽  
Lars Palm ◽  
Åke Lernmark ◽  
...  
Keyword(s):  
Single Cell ◽  
Single Cell Analysis ◽  
Author Response ◽  
Cell Analysis
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