scholarly journals Handheld Microflow Cytometer Based on a Motorized Smart Pipette, a Microfluidic Cell Concentrator, and a Miniaturized Fluorescence Microscope

Sensors ◽  
2019 ◽  
Vol 19 (12) ◽  
pp. 2761 ◽  
Author(s):  
Byeongyeon Kim ◽  
Dayoung Kang ◽  
Sungyoung Choi
Keyword(s):  
Flow Cytometry ◽  
High Throughput ◽  
Flow Cytometric Analysis ◽  
Sample Volume ◽  
Fluorescence Microscope ◽  
Comprehensive Approach ◽  
Optical Systems ◽  
Simple Protocol ◽  
Microflow Cytometer ◽  
On Chip

Miniaturizing flow cytometry requires a comprehensive approach to redesigning the conventional fluidic and optical systems to have a small footprint and simple usage and to enable rapid cell analysis. Microfluidic methods have addressed some challenges in limiting the realization of microflow cytometry, but most microfluidics-based flow cytometry techniques still rely on bulky equipment (e.g., high-precision syringe pumps and bench-top microscopes). Here, we describe a comprehensive approach that achieves high-throughput white blood cell (WBC) counting in a portable and handheld manner, thereby allowing the complete miniaturization of flow cytometry. Our approach integrates three major components: a motorized smart pipette for accurate volume metering and controllable liquid pumping, a microfluidic cell concentrator for target cell enrichment, and a miniaturized fluorescence microscope for portable flow cytometric analysis. We first validated the capability of each component by precisely metering various fluid samples and controlling flow rates in a range from 219.5 to 840.5 μL/min, achieving high sample-volume reduction via on-chip WBC enrichment, and successfully counting single WBCs flowing through a region of interrogation. We synergistically combined the three major components to create a handheld, integrated microflow cytometer and operated it with a simple protocol of drawing up a blood sample via pipetting and injecting the sample into the microfluidic concentrator by powering the motorized smart pipette. We then demonstrated the utility of the microflow cytometer as a quality control means for leukoreduced blood products, quantitatively analyzing residual WBCs (rWBCs) in blood samples present at concentrations as low as 0.1 rWBCs/μL. These portable, controllable, high-throughput, and quantitative microflow cytometric technologies provide promising ways of miniaturizing flow cytometry.

Quantitative determination of CD4/CD8 molecules by a cell marker ELISA

1994 ◽  
Vol 40 (1) ◽  
pp. 38-42 ◽  
Author(s):  
L Franke ◽  
E Nugel ◽  
W D Döcke ◽  
T Porstmann
Keyword(s):  
Flow Cytometry ◽  
T Lymphocytes ◽  
Flow Cytometric Analysis ◽  
Sample Volume ◽  
Small Sample ◽  
Cell Marker ◽  
Cell Counts ◽  
Flow Cytometric ◽  
A Cell

Abstract Determination of percentages of CD4+ and CD8+ T cells from patients with human immunodeficiency virus infection is usually done by flow cytometric analysis. We compared a cell marker ELISA with flow cytometry for quantitation of CD4 and CD8 molecules on T lymphocytes, and correlated the values both with the number of CD4+ and CD8+ T lymphocytes and with clinical data. Results by cell marker ELISA (y) correlated well with those by flow cytometric analysis (x); r = 0.69, P < 0.001 (y = 0.01x + 3.9) for CD4; r = 0.81, P < 0.001 (y = 0.03x + 5.4) for CD8; n = 343. The ELISA detected changes in numbers of CD8 molecules on the cells earlier than flow cytometry recognized changes in CD8+ T-cell counts. The advantages of the ELISA are the small sample volume required (0.5 mL of blood), its internal standardization by a CD4+/CD8+ cell line, and its simple and fast performance. The cell marker ELISA appears to be an efficient alternative to flow cytometry.

A deep learning-based concept for high throughput image flow cytometry

2021 ◽  
Vol 118 (12) ◽  
pp. 123701
Author(s):  
Julie Martin-Wortham ◽  
Steffen M. Recktenwald ◽  
Marcelle G. M. Lopes ◽  
Lars Kaestner ◽  
Christian Wagner ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Deep Learning ◽  
High Throughput ◽  
Image Flow

scholarly journals High-throughput multiparametric imaging flow cytometry: toward diffraction-limited sub-cellular detection and monitoring of sub-cellular processes

Cell Reports ◽  
2021 ◽  
Vol 34 (10) ◽  
pp. 108824
Author(s):  
Gregor Holzner ◽  
Bogdan Mateescu ◽  
Daniel van Leeuwen ◽  
Gea Cereghetti ◽  
Reinhard Dechant ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
High Throughput ◽  
Multiparametric Imaging ◽  
Cellular Processes ◽  
Imaging Flow Cytometry

scholarly journals A “No‐Touch” Antibody‐Staining Method of Adherent Cells for High‐Throughput Flow Cytometry in 384‐Well Microplate Format for Cell‐Based Drug Library Screening

2019 ◽  
Vol 97 (8) ◽  
pp. 845-851 ◽  
Author(s):  
Annelisa M. Cornel ◽  
Celina L. Szanto ◽  
Niek P. Til ◽  
Jeroen F. Velzen ◽  
Jaap J. Boelens ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
High Throughput ◽  
Staining Method ◽  
Adherent Cells ◽  
Library Screening ◽  
Antibody Staining

High-Throughput Raman Flow Cytometry and Beyond

2021 ◽  
Author(s):  
Julia Gala de Pablo ◽  
Matthew Lindley ◽  
Kotaro Hiramatsu ◽  
Keisuke Goda
Keyword(s):  
Flow Cytometry ◽  
High Throughput

scholarly journals Recent Bioinformatics Advances in the Analysis of High Throughput Flow Cytometry Data

2009 ◽  
Vol 2009 ◽  
pp. 1-2
Author(s):  
Raphael Gottardo ◽  
Ryan R. Brinkman ◽  
George Luta ◽  
Matt P. Wand
Keyword(s):  
Flow Cytometry ◽  
High Throughput ◽  
Flow Cytometry Data

scholarly journals Multiparameter Flow Cytometric Analysis of Antibiotic Effects on Membrane Potential, Membrane Permeability, and Bacterial Counts of Staphylococcus aureus andMicrococcus luteus

2000 ◽  
Vol 44 (4) ◽  
pp. 827-834 ◽  
Author(s):  
David J. Novo ◽  
Nancy G. Perlmutter ◽  
Richard H. Hunt ◽  
Howard M. Shapiro
Keyword(s):  
Staphylococcus Aureus ◽  
Flow Cytometry ◽  
Membrane Potential ◽  
Membrane Permeability ◽  
Micrococcus Luteus ◽  
Flow Cytometric Analysis ◽  
Bacterial Counts ◽  
Flow Cytometric ◽  
Permeability Parameter ◽  
Particle Counts

ABSTRACT Although flow cytometry has been used to study antibiotic effects on bacterial membrane potential (MP) and membrane permeability, flow cytometric results are not always well correlated to changes in bacterial counts. Using new, precise techniques, we simultaneously measured MP, membrane permeability, and particle counts of antibiotic-treated and untreated Staphylococcus aureus andMicrococcus luteus cells. MP was calculated from the ratio of red and green fluorescence of diethyloxacarbocyanine [DiOC2(3)]. A normalized permeability parameter was calculated from the ratio of far red fluorescence of the nucleic acid dye TO-PRO-3 and green DiOC2(3) fluorescence. Bacterial counts were calculated by the addition of polystyrene beads to the sample at a known concentration. Amoxicillin increased permeability within 45 min. At concentrations of <1 μg/ml, some organisms showed increased permeability but normal MP; this population disappeared after 4 h, while bacterial counts increased. At amoxicillin concentrations above 1 μg/ml, MP decreased irreversibly and the particle counts did not increase. Tetracycline and erythromycin caused smaller, dose- and time-dependent decreases in MP. Tetracycline concentrations of <1 μg/ml did not change permeability, while a tetracycline concentration of 4 μg/ml permeabilized 50% of the bacteria; 4 μg of erythromycin per ml permeabilized 20% of the bacteria. Streptomycin decreased MP substantially, with no effect on permeability; chloramphenicol did not change either permeability or MP. Erythromycin pretreatment of bacteria prevented streptomycin and amoxicillin effects. Flow cytometry provides a sensitive means of monitoring the dynamic cellular events that occur in bacteria exposed to antibacterial agents; however, it is probably simplistic to expect that changes in a single cellular parameter will suffice to determine the sensitivities of all species to all drugs.

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