scholarly journals Data-Driven Compensation for Flow Cytometry of Solid Tissues

2011 ◽  
Vol 2011 ◽  
pp. 1-11 ◽  
Author(s):  
Nickolaas Maria van Rodijnen ◽  
Math Pieters ◽  
Sjack Hoop ◽  
Marius Nap
Keyword(s):  
Flow Cytometry ◽  
Propidium Iodide ◽  
Dna Content ◽  
Flow Cytometer ◽  
Data Driven ◽  
Color Flow ◽  
Operator Experience ◽  
Solid Tissues

Propidium Iodide is a fluorochrome that is used to measure the DNA content of individual cells, taken from solid tissues, with a flow cytometer. Compensation for spectral cross-over of this fluorochrome still leads to compensation results that are depending on operator experience. We present a data-driven compensation (DDC) algorithm that is designed to automatically compensate combined DNA phenotype flow cytometry acquisitions. The generated compensation values of the DDC algorithm are validated by comparison with manually determined compensation values. The results show that (1) compensation of two-color flow cytometry leads to comparable results using either manual compensation or the DDC method; (2) DDC can calculate sample-specific compensation trace lines; (3) the effects of two different approaches to calculate compensation values can be visualized within one sample. We conclude that the DDC algorithm contributes to the standardization of compensation for spectral cross-over in flow cytometry of solid tissues.

scholarly journals Standardization of 8-color flow cytometry across different flow cytometer instruments: A feasibility study in clinical laboratories in Switzerland

2019 ◽  
Vol 475 ◽  
pp. 112348 ◽  
Author(s):  
Hana Glier ◽  
Ingmar Heijnen ◽  
Mathieu Hauwel ◽  
Jan Dirks ◽  
Stéphane Quarroz ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Feasibility Study ◽  
Flow Cytometer ◽  
Color Flow ◽  
Clinical Laboratories

scholarly journals Evolution of Genome Size in Duckweeds (Lemnaceae)

2011 ◽  
Vol 2011 ◽  
pp. 1-9 ◽  
Author(s):  
Wenqin Wang ◽  
Randall A. Kerstetter ◽  
Todd P. Michael
Keyword(s):  
Flow Cytometry ◽  
Genome Size ◽  
Propidium Iodide ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Continuous Increase ◽  
Spirodela Polyrhiza ◽  
Wolffia Arrhiza ◽  
Basic Reference

To extensively estimate the DNA content and to provide a basic reference for duckweed genome sequence research, the nuclear DNA content for 115 different accessions of 23 duckweed species was measured by flow cytometry (FCM) stained with propidium iodide as DNA stain. The 1C-value of DNA content in duckweed family varied nearly thirteen-fold, ranging from 150 megabases (Mbp) in Spirodela polyrhiza to 1,881 Mbp in Wolffia arrhiza. There is a continuous increase of DNA content in Spirodela, Landoltia, Lemna, Wolffiella, and Wolffia that parallels a morphological reduction in size. There is a significant intraspecific variation in the genus Lemna. However, no such variation was found in other studied species with multiple accessions of genera Spirodela, Landoltia, Wolffiella, and Wolffia.

Simplification of Clinical Flow Cytometry by Adoption of Premixed Antibody Panels

2020 ◽  
Vol 154 (Supplement_1) ◽  
pp. S112-S113
Author(s):  
J M Polski
Keyword(s):  
Flow Cytometry ◽  
Cost Effective ◽  
Standard Of Care ◽  
Flow Cytometer ◽  
Multiparameter Flow Cytometry ◽  
Complete Agreement ◽  
Color Flow ◽  
Additional Advantage ◽  
Medical Technologists ◽  
User Friendly

Abstract Introduction/Objective Multiparameter flow cytometry (MFC) is the standard of care for the diagnosis and monitoring of hematopoietic and lymphoid neoplasms. While MFC is a mature technology, new innovations are brought to the market. Such an innovation is the ClearLLab10C, FDA-cleared, 4-tube, 10-color, dry, premixed antibody panels (Beckman Coulter Diagnostics, Brea, California). Our laboratory decided to implement this new reagent system combined with 10-color clinical flow cytometer, Navios EX from the same vendor. This paper describes the clinical validation results for leukemia/lymphoma (LL) evaluation as well as logistical and labor savings in the laboratory. Methods Thirty specimens submitted for LL evaluations were tested by our existing 5-color flow cytometer with custom antibody combinations as well as by the Navios EX instrument with the ClearLLab10C product supplemented by additional antibody combinations for intracellular analysis. The validation cases included normal and abnormal cases, representing bone marrows, blood, lymph nodes, and a few other specimens. Results There was a complete agreement in the qualitative results. The new simplified premixed reagents allow for noticeable simplification of inventory and saving in labor. The existing LL antibody cocktails required stocking 41 antibodies. The new platform requires only 4 LL reagents for surface immunophenotyping and we elected to stock additional 8 antibodies for optional intracellular evaluation. The amount of manual pipetting is greatly reduced. One additional advantage of utilizing the new ClearLLab10C is the available electronic and book publication from the vendor illustrating typical results for 24 normal and abnormal specimens analyzed using ClearLLab10C. This resource together with a user-friendly analysis software Kaluza C greatly facilitate interpretation of 10-color results by pathologists, medical technologists, and especially by the pathologists in training. Conclusion ClearLLab10C product is a cost-effective and user-friendly solution for laboratories seeking a streamlined MFC and a great reduction of inventory and labor.

scholarly journals Energy Transfer between Fluorescein Isothiocyanate and Propidium Iodide – A Problem in the Estimation of Tpotwith the Bromodeoxyuridine–DNA Flow Cytometry Technique?

1999 ◽  
Vol 19 (2) ◽  
pp. 91-98 ◽  
Author(s):  
Maria C. Johansson ◽  
Bo Baldetorp ◽  
Stina M. Oredsson
Keyword(s):  
Flow Cytometry ◽  
Energy Transfer ◽  
Propidium Iodide ◽  
Dna Content ◽  
Fluorescein Isothiocyanate ◽  
Dna Flow Cytometry ◽  
Synthesis Time ◽  
Close Proximity ◽  
Potential Doubling Time ◽  
Potential Doubling

Energy transfer in flow cytometry can occur when two fluorochromes are bound in close proximity (generally within 100 Å) and the emission spectrum of one fluorochrome overlaps significantly with the excitation spectrum of the other. The latter criterium is fullfilled for the fluorochromes fluorescein isothiocyanate and propidium iodide and also the former when they, e.g., are used in bromodeoxyuridine – DNA flow cytometry methods. In the present growth kinetic study using this method, we show that energy transfer does take place between fluorescein isothiocyanate and propidium iodide which results in a detected increase in DNA content with 2–3%. Despite the erroneous increase in the obtained DNA content values, this does not seem to have any influence on the calculation of DNA synthesis time and potential doubling time where the DNA content, based on the relative movement principle of the labelled cells, is used.

Novel Methodology for the Detection of Cryptosporidium parvum: A Comparison of Cooled Charge Couple Devices (CCD) and Flow Cytometry

1993 ◽  
Vol 27 (3-4) ◽  
pp. 89-92 ◽  
Author(s):  
Andrew Campbell ◽  
Lucy Robertson ◽  
Huw Smith
Keyword(s):  
Monoclonal Antibody ◽  
Flow Cytometry ◽  
Cryptosporidium Parvum ◽  
Propidium Iodide ◽  
Three Dimensional ◽  
Flow Cytometer ◽  
Cryptosporidium Oocysts ◽  
Exact Size

Flow cytometry and CCD were assessed for their usefialness in the detection of oocysts of Cryptosporidium. Oocysts were labelled with FITC-monoclonal antibody and with the nuclear stains 4’6-diamidino-2-phenyl indole (DAPI) and propidium iodide (PI) before analysis by flow cytometer and CCD. Although the flow cytometer tested was able to concentrate particles and place them on a slide for subsequent viewing, readily sorting oocysts from contaminating debris, confirmation by fluoresence microscopy was still essential, even when additional parameters such as the inclusion of DAPI is used. Initial observations from the use of CCD, however, suggested that screening samples for oocysts was a possibility. Three dimensional visualisation of individual oocysts can map precisiely both the detailed morphology and the exact size of oocysts, thereby making confirmation by fluoresence microscopy unneccesary.

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