Flow cytometer in the infrared: Inexpensive modifications to a commercial instrument

2005 ◽  
Vol 67A (2) ◽  
pp. 104-111 ◽  
Author(s):  
Carleton C. Stewart ◽  
Mitchell L. Woodring ◽  
Edward Podniesinski ◽  
Brian Gray
Keyword(s):  
Flow Cytometer ◽  
Commercial Instrument

Electrostatic optics and aberration correction in the 1940s and today

1992 ◽  
Vol 50 (1) ◽  
pp. 6-7
Author(s):  
Gertrude F. Rempfer
Keyword(s):  
Electron Microscope ◽  
Focal Point ◽  
Focal Length ◽  
High Vacuum ◽  
Electron Optics ◽  
Applied Physics ◽  
Electrostatic Lenses ◽  
Commercial Instrument ◽  
The University ◽  
Theory And Experiment

I became involved in electron optics in early 1945, when my husband Robert and I were hired by the Farrand Optical Company. My husband had a mathematics Ph.D.; my degree was in physics. My main responsibilities were connected with the development of an electrostatic electron microscope. Fortunately, my thesis research on thermionic and field emission, in the late 1930s under the direction of Professor Joseph E. Henderson at the University of Washington, provided a foundation for dealing with electron beams, high vacuum, and high voltage.At the Farrand Company my co-workers and I used an electron-optical bench to carry out an extensive series of tests on three-electrode electrostatic lenses, as a function of geometrical and voltage parameters. Our studies enabled us to select optimum designs for the lenses in the electron microscope. We early on discovered that, in general, electron lenses are not “thin” lenses, and that aberrations of focal point and aberrations of focal length are not the same. I found electron optics to be an intriguing blend of theory and experiment. A laboratory version of the electron microscope was built and tested, and a report was given at the December 1947 EMSA meeting. The micrograph in fig. 1 is one of several which were presented at the meeting. This micrograph also appeared on the cover of the January 1949 issue of Journal of Applied Physics. These were exciting times in electron microscopy; it seemed that almost everything that happened was new. Our opportunities to publish were limited to patents because Mr. Farrand envisaged a commercial instrument. Regrettably, a commercial version of our laboratory microscope was not produced.

Atomic Spectroscopy in the FIM

1986 ◽  
Vol 44 ◽  
pp. 758-761
Author(s):  
J. J. Hren ◽  
S. D. Walck
Keyword(s):  
Electron Microscope ◽  
Electric Fields ◽  
Atom Probe ◽  
Atomic Spectroscopy ◽  
Single Atom ◽  
Statistical Sampling ◽  
Commercial Instrument ◽  
Available Technology ◽  
High Electric Fields ◽  
Field Ion Microscope

The field ion microscope (FIM) has had the ability to routinely image the surface atoms of metals since Mueller perfected it in 1956. Since 1967, the TOF Atom Probe has had single atom sensitivity in conjunction with the FIM. “Why then hasn't the FIM enjoyed the success of the electron microscope?” The answer is closely related to the evolution of FIM/Atom Probe techniques and the available technology. This paper will review this evolution from Mueller's early discoveries, to the development of a viable commercial instrument. It will touch upon some important contributions of individuals and groups, but will not attempt to be all inclusive. Variations in instrumentation that define the class of problems for which the FIM/AP is uniquely suited and those for which it is not will be described. The influence of high electric fields inherent to the technique on the specimens studied will also be discussed. The specimen geometry as it relates to preparation, statistical sampling and compatibility with the TEM will be examined.

Loss of Thrombin-Induced Ca2+ Mobilization in a Subpopulation of Platelets during Storage

1991 ◽  
Vol 66 (03) ◽  
pp. 350-354 ◽  
Author(s):  
Rob Fijnheer ◽  
Christa H E Homburg ◽  
Berend Hooibrink ◽  
Martine N Boomgaard ◽  
Dirk de Korte ◽  
...  
Keyword(s):  
Human Platelets ◽  
Flow Cytometer ◽  
Maximal Concentration ◽  
Gradual Decline ◽  
Platelet Storage ◽  
Functional Defect ◽  
Total Fluorescence ◽  
Induced Changes

SummaryThrombin-induced changes in cytosolic free Ca2+ ([Ca2+]i) were studied in human platelets that had been stored for up to 6 days. Changes in [Ca2+]i were measured with Indo-1-loaded platelets and quantitated with two different methods: (i) measurement of the changes in total fluorescence; (ii) measurement of the [Ca2+]i changes in individual platelets in a flow cytometer, allowing the detection of non-responding platelets. The maximal concentration of [Ca2+]i after stimulation with 0.5 U of thrombin/ml decreased from 544 ± 58 nM (mean ± SEM, n = 6) on day 0, to 276 ± 9 nM on day 3 and to 203 ± 23 nM on day 6. The percentage of platelets responding to 0.5 U of thrombin/ml declined from 90 ± 2% on day 0 to 72 ± 4% on day 3, and to 47 ± 8% on day 6. Nevertheless, also the responding platelets showed a decreased rise in [Ca2+]i.The study shows that during platelet storage a decrease in the rise in [Ca2+]i upon thrombin stimulation occurs. This decrease is partly due to the formation of a subpopulation of platelets that is completely unresponsive and partly due to a decreased responsiveness in the remainder of the platelets; it is not due to a gradual decline in [Ca2+]i rise in all platelets. This phenomenon provides new insight in the functional defect of stored platelets.

Identification of antibiotics in meat. Flow cytometry capabilities

Vsyo o myase ◽  
2020 ◽  
pp. 56-60
Author(s):  
Zayko E.V. ◽  
◽  
Kuznetsova O.A. ◽  
Bataeva D.S. ◽  
Grudistova M.A. ◽  
...  
Keyword(s):  
Raw Materials ◽  
Animal Husbandry ◽  
Flow Cytometer ◽  
Poultry Meat ◽  
Meat Processing ◽  
Beta Lactams ◽  
Antimicrobial Substances ◽  
Chemotherapy Drugs ◽  
Processing Plants ◽  
Meat Samples

The problem of the uncontrolled use of antibiotics currently remains unresolved. Step-by-step monitoring of meat using modern methods will reduce the risk of using contaminated meat raw materials for food production. Qualitative monitoring will identify samples containing residual antimicrobial substances. The use of methods for identifying groups of antibiotics will help narrow the search for antibiotics by expensive chromatographic methods. A study was carried out of beef, pork and poultry meat, which is used in meat processing plants in the production of raw smoked sausages, using two methods. At the first stage, using a qualitative microbiological method, the raw meat was evaluated for the presence of antimicrobial substances, then their group was determined using a NovoCyte flow cytometer. According to the results of a study on a flow cytometer, it was found that out of 10 groups of antibiotics that can be determined by the tested method, the group of lincosamides was not found in all meat samples. The most common groups of chemotherapy drugs in pork were sulfonamides – 29.6 %, tetracycline group – 18.5 % and beta-lactams – 14.8 %, and in beef aminoglycosides – 36.7 %, phenicols – 30 % and beta-lactams – 13.3 %. In poultry meat samples, the most common were sulfonamides – 23.2 %, fenicols – 23.2 %, and beta-lactams – 16 %. Five groups of antibiotics were found in all studied types of meat: fenicols, β-lactams, macrolides, polypeptide antibiotics, and quinolones. This indicates their widespread use in animal husbandry and poultry farming.

scholarly journals Single Step In Situ Detection of Surface Protein and MicroRNA in Clustered Extracellular Vesicles Using Flow Cytometry

2021 ◽  
Vol 10 (2) ◽  
pp. 319
Author(s):  
Hee Cheol Yang ◽  
Won Jong Rhee
Keyword(s):  
Extracellular Vesicles ◽  
Liquid Biopsy ◽  
Molecular Beacon ◽  
Disease Diagnosis ◽  
Single Step ◽  
Flow Cytometer ◽  
Surface Proteins ◽  
Biomarker Detection ◽  
In Situ Detection

Because cancers are heterogeneous, it is evident that multiplexed detection is required to achieve disease diagnosis with high accuracy and specificity. Extracellular vesicles (EVs) have been a subject of great interest as sources of novel biomarkers for cancer liquid biopsy. However, EVs are nano-sized particles that are difficult to handle; thus, it is necessary to develop a method that enables efficient and straightforward EV biomarker detection. In the present study, we developed a method for single step in situ detection of EV surface proteins and inner miRNAs simultaneously using a flow cytometer. CD63 antibody and molecular beacon-21 were investigated for multiplexed biomarker detection in normal and cancer EVs. A phospholipid-polymer-phospholipid conjugate was introduced to induce clustering of the EVs analyzed using nanoparticle tracking analysis, which enhanced the detection signals. As a result, the method could detect and distinguish cancer cell-derived EVs using a flow cytometer. Thus, single step in situ detection of multiple EV biomarkers using a flow cytometer can be applied as a simple, labor- and time-saving, non-invasive liquid biopsy for the diagnosis of various diseases, including cancer.

scholarly journals Sources of Variability in the Response of Labeled Microspheres and B Cells during the Analysis by a Flow Cytometer

2021 ◽  
Vol 22 (15) ◽  
pp. 8256
Author(s):  
Adolfas K. Gaigalas ◽  
Yu-Zhong Zhang ◽  
Linhua Tian ◽  
Lili Wang
Keyword(s):  
B Cells ◽  
Laser Beam ◽  
Cross Section ◽  
Mean Fluorescence Intensity ◽  
Numerical Aperture ◽  
Flow Cytometer ◽  
Fluorescence Signal ◽  
Minor Importance ◽  
Reliable Measure ◽  
The Mean

A stochastic model of the flow cytometer measurement process was developed to assess the nature of the observed coefficient of variation (CV%) of the mean fluorescence intensity (MFI) from a population of labeled microspheres (beads). Several sources of variability were considered: the total number of labels on a bead, the path through the laser beam, the optical absorption cross-section, the quantum yield, the numerical aperture of the collection optics, and the photoelectron conversion efficiency of the photomultiplier (PMT) cathode. The variation in the number of labels on a bead had the largest effect on the CV% of the MFI of the bead population. The variation in the path of the bead through the laser beam was minimized using flat-top lasers. The variability in the average optical properties of the labels was of minor importance for beads with sufficiently large number of labels. The application of the bead results to the measured CV% of labeled B cells indicated that the measured CV% was a reliable measure of the variability of antibodies bound per cell. With some modifications, the model can be extended to multicolor flow cytometers and to the study of CV% from cells with low fluorescence signal.

scholarly journals Cord Blood Extracellular Vesicles Analyzed by Flow Cytometry with Thresholding Using 405 nm or 488 nm Laser Leads to Concurrent Results

Diagnostics ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 1320
Author(s):  
Kristýna Pekárková ◽  
Jakub Soukup ◽  
Marie Kostelanská ◽  
Jan Širc ◽  
Zbyněk Straňák ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Cord Blood ◽  
Extracellular Vesicles ◽  
Correlation Coefficients ◽  
Flow Cytometer ◽  
Liquid Biopsies ◽  
Physiological Conditions ◽  
Flow Cytometric ◽  
Term Births ◽  
And Control

Extracellular vesicles (EVs) from liquid biopsies are extensively analyzed by flow cytometry, a technology that is continuously evolving. Thresholding utilizing a violet 405 nm laser side scatter (VSSC) has recently been implemented. Here, we collected set of large EV (lEV) samples from cord blood, which we analyzed using a standard flow cytometer improved via a 405 nm laser side scatter. Samples were analyzed using two distinct thresholding methods—one based on VSSC, and one based on VSSC combined with fluorescence thresholding on stained phosphatidylserine. Through these thresholding methods, we compared lEVs from pre-term births and control cord blood. Double-labeled lEVs with platelet CD36+/CD41+, activated platelet CD41+/CD62P+ and endothelial CD31+/CD105+ antibodies were used. Apart from comparing the two groups together, we also correlated measured lEVs with the thresholding methods. We also correlated the results of this study with data analyzed in our previous study in which we used a conventional 488 nm laser SSC. We did not find any difference between the two cord blood groups. However, we found highly concurrent data via our correlation of the thresholding methods, with correlation coefficients ranging from 0.80 to 0.96 even though the numbers of detected lEVs differed between thresholding methods. In conclusion, our approaches to thresholding provided concurrent data and it seems that improving the cytometer with the use of a VSSC increases its sensitivity, despite not being particularly critical to the validity of flow cytometric studies that compare pathological and physiological conditions in liquid biopsies.

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