scholarly journals Application of flow cytometry and cell sorting to megakaryocytopoiesis

Blood ◽  
1979 ◽  
Vol 53 (4) ◽  
pp. 732-745 ◽  
Author(s):  
A Nakeff ◽  
F Valeriote ◽  
JW Gray ◽  
RJ Grabske
Keyword(s):  
Flow Cytometry ◽  
Cell Sorting ◽  
Significant Advance ◽  
Glass Slides ◽  
Acetylcholinesterase Staining ◽  
Fcm Analysis ◽  
Cell Fractions ◽  
Kinetics Of ◽  
Supravital Staining

Abstract We have employed flow cytometry (FCM) and cell sorting to quantitate and study megakaryocytes in mouse and rat femoral marrow following their 20- to 30-fold concentration by centrifugal elutriation (CE). This enrichment of megakaryocytes permitted the first determination of their DNA-related fluorescence by FCM analysis following DNA staining. Fluorescence distributions of CE-enriched cell fractions following supravital staining with Hoechst 33342 were similar to those following chromomycin A3 staining of ethanol-fixed cells. Microscopic examination of cells sorted onto glass slides on the basis of their DNA-related fluorescence following supravital staining together with specific acetylcholinesterase staining for megakaryocytes indicated that megakaryocytes generally increased in cell size with increasing DNA content. This technologic application represents a significant advance in the study of megakaryocytopoiesis, since the kinetics of either the normal or perturbed population can now be studied rapidly and quantitatively.

scholarly journals Application of flow cytometry and cell sorting to megakaryocytopoiesis

Blood ◽  
1979 ◽  
Vol 53 (4) ◽  
pp. 732-745 ◽  
Author(s):  
A Nakeff ◽  
F Valeriote ◽  
JW Gray ◽  
RJ Grabske
Keyword(s):  
Flow Cytometry ◽  
Cell Sorting ◽  
Significant Advance ◽  
Glass Slides ◽  
Acetylcholinesterase Staining ◽  
Fcm Analysis ◽  
Cell Fractions ◽  
Kinetics Of ◽  
Supravital Staining

We have employed flow cytometry (FCM) and cell sorting to quantitate and study megakaryocytes in mouse and rat femoral marrow following their 20- to 30-fold concentration by centrifugal elutriation (CE). This enrichment of megakaryocytes permitted the first determination of their DNA-related fluorescence by FCM analysis following DNA staining. Fluorescence distributions of CE-enriched cell fractions following supravital staining with Hoechst 33342 were similar to those following chromomycin A3 staining of ethanol-fixed cells. Microscopic examination of cells sorted onto glass slides on the basis of their DNA-related fluorescence following supravital staining together with specific acetylcholinesterase staining for megakaryocytes indicated that megakaryocytes generally increased in cell size with increasing DNA content. This technologic application represents a significant advance in the study of megakaryocytopoiesis, since the kinetics of either the normal or perturbed population can now be studied rapidly and quantitatively.

scholarly journals KD determination from time-resolved experiments on live cells with LigandTracer and reconciliation with end-point flow cytometry measurements

2021 ◽  
Author(s):  
Diana Spiegelberg ◽  
Jonas Stenberg ◽  
Pascale Richalet ◽  
Marc Vanhove
Keyword(s):  
Flow Cytometry ◽  
Live Cells ◽  
Time Resolved ◽  
Surface Receptors ◽  
Full Characterization ◽  
End Point ◽  
Titration Curves ◽  
Kinetics Of

AbstractDesign of next-generation therapeutics comes with new challenges and emulates technology and methods to meet them. Characterizing the binding of either natural ligands or therapeutic proteins to cell-surface receptors, for which relevant recombinant versions may not exist, represents one of these challenges. Here we report the characterization of the interaction of five different antibody therapeutics (Trastuzumab, Rituximab, Panitumumab, Pertuzumab, and Cetuximab) with their cognate target receptors using LigandTracer. The method offers the advantage of being performed on live cells, alleviating the need for a recombinant source of the receptor. Furthermore, time-resolved measurements, in addition to allowing the determination of the affinity of the studied drug to its target, give access to the binding kinetics thereby providing a full characterization of the system. In this study, we also compared time-resolved LigandTracer data with end-point KD determination from flow cytometry experiments and hypothesize that discrepancies between these two approaches, when they exist, generally come from flow cytometry titration curves being acquired prior to full equilibration of the system. Our data, however, show that knowledge of the kinetics of the interaction allows to reconcile the data obtained by flow cytometry and LigandTracer and demonstrate the complementarity of these two methods.

scholarly journals GENOME SIZE DETERMINATION OF CUCUMBER (CUCUMIS SATIVUS), HONEYDEW (CUCUMIS MELO INODORUS) AND ROCK MELON (CUCUMIS MELO CANTALUPENSIS) VIA FLOW CYTOMETRY

2021 ◽  
Vol 5 (1) ◽  
pp. 14-16
Author(s):  
Raden Muhamad Imaduddin Yumni ◽  
Mohd Fauzihan Karim ◽  
Mohd Razik Midin
Keyword(s):  
Flow Cytometry ◽  
Genome Size ◽  
Cucumis Melo ◽  
As Species ◽  
External Reference ◽  
The Family ◽  
Cucumis Species ◽  
Fcm Analysis ◽  
Interspecific And Intraspecific Variation

The family of Cucurbitaceae consists of species with economical and nutritional value. Morphologically, there are only few differences between Cucumis species. The interspecific and intraspecific variation in the genome size of the Cucumis species are not discovered yet. Due to this, this study aims to determine the genome size of C. sativus, C. melo inodorus and C. melo cantalupensis using flow cytometry (FCM) method. Nuclei suspension of selected Cucumis species were extracted using LBO1 lysis buffer by manual chopping technique and stained by propidium iodide priot to FCM analysis. Genome size of C. sativus, C. melo inodorus (Honeydew) and C. melo cantalupensis (Rockmelon) were determined by using Glycine max (Soybean) as an external reference standard (2C = 2.5 pg). This study found that the genome size of C. sativus, C. melo inodorus and C. melo cantalupensis estimated to be 2.83 pg, 3.00 pg and 3.47 pg respectively. The genome size data obtained from this study can be used in future genome studies as well as species characterization.

scholarly journals Nuclear Genome Size Determination Of Christia Vespertilionis Via Flow Cytometry

2020 ◽  
Vol 4 (2) ◽  
pp. 72-75
Author(s):  
Mohd Razik Midin ◽  
Muhammad Irfan Fikri ◽  
Siti Sarah Zailani
Keyword(s):  
Flow Cytometry ◽  
Genome Size ◽  
Nuclear Genome ◽  
Size Determination ◽  
Size Estimation ◽  
Butterfly Wing ◽  
External Reference ◽  
Fcm Analysis ◽  
Suspension Preparation

AbstractChristia vespertilionis (butterfly wing plant) is an ornamental plant originated from South East Asia with reported usage in traditional medicine practice and potential as an anticancer and antitumor. This research aims to estimate the genome size of C. vespertilionis via flow cytometry (FCM) method. The research was conducted with the optimisation of nuclear suspension preparation followed by the genome size estimation. Two chopping techniques [manual chopping (MC) and BDTM Medimachine (MM)] and two lysis buffers (Otto and LBO1) were tested. Otto buffer with manual chopping was found to be the most suitable method, generated fine DNA peak with minimum debris background, and coefficient of variation (CV) value less than 3%. Five replicates of the FCM analysis were made for the genome size determination. The estimated genome size of C. vespertilionis was found to be 3.22 pg by using Glycine max cv. Polanka (2C=2.5pg) as an external reference standard. Further comparison with other Christia species was not possible due to the lack of data on genome size. The genome size data of C. vespertilionis can be useful for future morphology and genetics studies of Christia species.

Determination of microbiological activity during the processing of frost damaged sugar beets

2014 ◽  
pp. 228-231 ◽  
Author(s):  
Maciej Wojtczak ◽  
Aneta Antczak-Chrobot ◽  
Edyta Chmal-Fudali ◽  
Agnieszka Papiewska
Keyword(s):  
Sugar Beet ◽  
Microbiological Activity ◽  
Microbiological Contamination ◽  
Sugar Beets ◽  
Bacterial Metabolites ◽  
Kinetics Of

The aim of the study is to evaluate the kinetics of the synthesis of dextran and other bacterial metabolites as markers of microbiological contamination of sugar beet.

Studies on the Behavior Some Paints in Electro-insulating Fluid Based on Vegetable Esters

2018 ◽  
Vol 69 (5) ◽  
pp. 1139-1144
Author(s):  
Iosif Lingvay ◽  
Adriana Mariana Bors ◽  
Livia Carmen Ungureanu ◽  
Valerica Stanoi ◽  
Traian Rus
Keyword(s):  
Structural Changes ◽  
Oxidation Processes ◽  
High Oleic ◽  
Painting Materials ◽  
Insulating Paper ◽  
Different Types ◽  
Mechanism And Kinetics ◽  
Kinetics Of ◽  
Natural Ester

For the purpose of using three different types of painting materials for the inner protection of the transformer vats, their behavior was studied under actual conditions of operation in the transformer (thermal stress in electro-insulating fluid based on the natural ester in contact with copper for electro-technical use and electro-insulating paper). By comparing determination of the content in furans products (HPLC technique) and gases formed (by gas-chromatography) in the electro-insulating fluid (natural ester with high oleic content) thermally aged at 130 �C to 1000 hours in closed glass vessels, it have been found that the presence the investigated painting materials lead to a change in the mechanism and kinetics of the thermo-oxidation processes. These changes are supported by oxygen dissolved in oil, what leads to decrease both to gases formation CO2, CO, H2, CH4, C2H4 and C2H6) and furans products (5-HMF, 2-FOL, 2 -FAL and 2-ACF). The painting materials investigated during the heat treatment applied did not suffer any remarkable structural changes affecting their functionality in the electro-insulating fluid based on vegetable esters.

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