scholarly journals Non-invasive cell counting of adherent, suspended and encapsulated mammalian cells using optical density

BioTechniques ◽  
2020 ◽  
Vol 68 (1) ◽  
pp. 35-40
Author(s):  
Ayesha Aijaz ◽  
Dylan Trawinski ◽  
Scott McKirgan ◽  
Biju Parekkadan
Keyword(s):  
Cell Count ◽  
Mammalian Cell ◽  
Optical Density ◽  
Mammalian Cells ◽  
Cell Number ◽  
Cell Counting ◽  
Non Invasive ◽  
Mammalian Cell Growth ◽  
Polymeric Microcapsules ◽  
Invasive Cell

In situ measurement to determine mammalian cell number in a non-invasive, non-destructive and reagent-free manner is needed to enable continuous cell manufacturing. An analytical method is presented for non-invasive cell counting by conducting multiwavelength spectral analysis of mammalian cells achieving a minimal detectable cell count of 62,500 at 295 nm. Light absorbance was insensitive to culture volume, giving an absolute cell count rather than a concentration. The activation state of cells was also considered. The study was extended to quantification within polymeric microcapsules as an advanced substrate for mammalian cell growth in bioreactor formats and resulted in an offset directly correlating with the absorbance maxima of the polymer. These studies provide feasibility for optical density as a simple end point to indirectly quantify mammalian cell number for continuous monitoring of cell cultures.

NanoLiterBioReactor: Monitoring of Long-Term Mammalian Cell Physiology at Nanofabricated Scale

2004 ◽  
Vol 820 ◽  
Author(s):  
Ales Prokop ◽  
Zdenka Prokop ◽  
David Schaffer ◽  
Eugene Kozlov ◽  
John Wikswo ◽  
...  
Keyword(s):  
Cell Lines ◽  
Mammalian Cell ◽  
Mammalian Cells ◽  
Cellular Response ◽  
Cell Counting ◽  
Cell Physiology ◽  
Mammalian Cell Lines ◽  
Cell Counts ◽  
Multiple Cell

AbstractThere is a need for microminiaturized cell-culture environments, i.e., NanoLiter BioReactors (NBRs), for growing and maintaining populations of up to several hundred cultured mammalian cells in volumes three orders of magnitude smaller than those contained in standard multi-well screening plates. Reduced NBR volumes would not only shorten the time required for diffusive mixing, for achieving thermal equilibrium, and for cells to grow to confluence, but also simplify accurate cell counting, minimize required volumes of expensive analytical pharmaceuticals or toxins, and allow for thousands of culture chambers on a single instrumented chip. These devices would enable the development of a new class of miniature, automated cell-based bioanalysis arrays for monitoring the immediate environment of multiple cell lines and assessing the effects of drug or toxin exposure. The challenge, beyond that of optimizing the NBR physically, is to detect cellular response, provide appropriate control signals, and, eventually, facilitate closed-loop adjustments of the environment--e.g., to control temperature, pH, ionic concentration, etc., to maintain homeostasis, or to apply drugs or toxins followed by the adaptive administration of a selective toxin antidote. To characterize in a nonspecific manner the metabolic activity of cells, the biosensor elements of the NBR might include planar pH, dissolved oxygen, and redox potential sensors, or even an isothermal picocalorimeter (pC) to monitor thermodynamic response. Equipped with such sensors, the NBR could be used to perform short- and long-term cultivation of several mammalian cell lines in a perfused system, and to monitor their response to analytes in a massively parallel format. This approach will enable automated, parallel, and multiphasic monitoring of multiple cell lines for drug and toxicology screening. An added bonus is the possibility of studying cell populations with low cell counts whose constituents are completely detached from typical tissue environment, or populations in controlled physical and chemical gradients.

NanoLiterBioReactor: Monitoring of Long-Term Mammalian Cell Physiology at Nanofabricated Scale

2004 ◽  
Vol 823 ◽  
Author(s):  
Ales Prokop ◽  
Zdenka Prokop ◽  
David Schaffer ◽  
Eugene Kozlov ◽  
John Wikswo ◽  
...  
Keyword(s):  
Cell Lines ◽  
Mammalian Cell ◽  
Mammalian Cells ◽  
Cellular Response ◽  
Cell Counting ◽  
Cell Physiology ◽  
Mammalian Cell Lines ◽  
Cell Counts ◽  
Multiple Cell

AbstractThere is a need for microminiaturized cell-culture environments, i.e., NanoLiter BioReactors (NBRs), for growing and maintaining populations of up to several hundred cultured mammalian cells in volumes three orders of magnitude smaller than those contained in standard multi-well screening plates. Reduced NBR volumes would not only shorten the time required for diffusive mixing, for achieving thermal equilibrium, and for cells to grow to confluence, but also simplify accurate cell counting, minimize required volumes of expensive analytical pharmaceuticals or toxins, and allow for thousands of culture chambers on a single instrumented chip. These devices would enable the development of a new class of miniature, automated cell- based bioanalysis arrays for monitoring the immediate environment of multiple cell lines and assessing the effects of drug or toxin exposure. The challenge, beyond that of optimizing the NBR physically, is to detect cellular response, provide appropriate control signals, and, eventually, facilitate closed-loop adjustments of the environment–e.g., to control temperature, pH, ionic concentration, etc., to maintain homeostasis, or to apply drugs or toxins followed by the adaptive administration of a selective toxin antidote. To characterize in a nonspecific manner the metabolic activity of cells, the biosensor elements of the NBR might include planar pH, dissolved oxygen, and redox potential sensors, or even an isothermal picocalorimeter (pC) to monitor thermodynamic response. Equipped with such sensors, the NBR could be used to perform short- and long-term cultivation of several mammalian cell lines in a perfused system, and to monitor their response to analytes in a massively parallel format. This approach will enable automated, parallel, and multiphasic monitoring of multiple cell lines for drug and toxicology screening. An added bonus is the possibility of studying cell populations with low cell counts whose constituents are completely detached from typical tissue environment, or populations in controlled physical and chemical gradients.

scholarly journals Physicochemical Investigation of Biosynthesis of a Protein Coating on Glass That Promotes Mammalian Cell Growth Using Lactobacillus rhamnosus GG Bacteria

Coatings ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 1410
Author(s):  
Kamil Kaminski ◽  
Karolina Syrek ◽  
Joanna Grudzień ◽  
Magdalena Obloza ◽  
Monika Adamczyk ◽  
...  
Keyword(s):  
Cell Growth ◽  
Mammalian Cell ◽  
Mammalian Cells ◽  
Ex Vivo ◽  
Lactobacillus Rhamnosus ◽  
Vero Cells ◽  
Lactobacillus Rhamnosus Gg ◽  
Composition And Structure ◽  
Mammalian Cell Growth ◽  
Protein Components

Glass surfaces, although the first to be used for culturing ex vivo adherent cells, are not the perfect substrates for this purpose. Today, plastics dominate these applications, but in light of the global trend to reduce the use of synthetic polymers, it is reasonable to consider a return to glass vessels with coatings for these purposes. The ideal surface for cell growth is one that simulates the composition and structure of the mainly protein-based intercellular matrix. The work presented here shows a new idea of preparing porous protein coatings on glass using biosynthesis. The process utilizes the colonization of the gold nanoparticle-coated glass surface with Lactobacillus rhamnosus GG bacteria, followed by permeabilization (using ethanol) of their membrane and partial thermal degradation (at 160 °C in vacuum) of the surface-bound protein components of these microorganisms. It results in a development of coating on the glass that promotes mammalian cell growth, which has been preliminary confirmed using Vero cells. Subsequent steps in the formation of coating components were documented by reflectance ultraviolet and visible spectra and infrared spectroscopy. The presence of microorganisms and mammalian cells was confirmed using scanning electron and optical microscopy and crystalline violet staining.

Apoptosis and development

2003 ◽  
Vol 39 ◽  
pp. 11-24 ◽  
Author(s):  
Justin V McCarthy
Keyword(s):  
Drosophila Melanogaster ◽  
Mammalian Cells ◽  
Cell Number ◽  
Model Organisms ◽  
Biological Processes ◽  
Nematode Caenorhabditis Elegans ◽  
Apoptotic Pathway ◽  
Genetic Pathways ◽  
Evolutionarily Conserved ◽  
Multicellular Organisms

Apoptosis is an evolutionarily conserved process used by multicellular organisms to developmentally regulate cell number or to eliminate cells that are potentially detrimental to the organism. The large diversity of regulators of apoptosis in mammalian cells and their numerous interactions complicate the analysis of their individual functions, particularly in development. The remarkable conservation of apoptotic mechanisms across species has allowed the genetic pathways of apoptosis determined in lower species, such as the nematode Caenorhabditis elegans and the fruitfly Drosophila melanogaster, to act as models for understanding the biology of apoptosis in mammalian cells. Though many components of the apoptotic pathway are conserved between species, the use of additional model organisms has revealed several important differences and supports the use of model organisms in deciphering complex biological processes such as apoptosis.

AlGaN/GaN heterostructures for non-invasive cell electrophysiological measurements

2007 ◽  
Vol 23 (4) ◽  
pp. 513-519 ◽  
Author(s):  
Jinjiang Yu ◽  
Shrawan Kumar Jha ◽  
Lidan Xiao ◽  
Qingjun Liu ◽  
Ping Wang ◽  
...  
Keyword(s):  
Non Invasive ◽  
Electrophysiological Measurements ◽  
Invasive Cell

scholarly journals The Graphical Representation of Cell Count Representation; a New Procedure for the Diagnosis of Periprosthetic Joint Infections

Antibiotics ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 346
Author(s):  
Bernd Fink ◽  
Marius Hoyka ◽  
Elke Weissbarth ◽  
Philipp Schuster ◽  
Irina Berger
Keyword(s):  
Cell Count ◽  
Graphical Representation ◽  
Diagnostic Value ◽  
Cell Counting ◽  
Wear Particles ◽  
Type Iv ◽  
Joint Infections ◽  
Cell Counts ◽  
Matrix Assessment ◽  
Wbc Count

Aim: This study was designed to answer the question whether a graphical representation increase the diagnostic value of automated leucocyte counting of the synovial fluid in the diagnosis of periprosthetic joint infections (PJI). Material and methods: Synovial aspirates from 322 patients (162 women, 160 men) with revisions of 192 total knee and 130 hip arthroplasties were analysed with microbiological cultivation, determination of cell counts and assay of the biomarker alpha-defensin (170 cases). In addition, microbiological and histological analysis of the periprosthetic tissue obtained during the revision surgery was carried out using the ICM classification and the histological classification of Morawietz and Krenn. The synovial aspirates were additionally analysed to produce dot plot representations (LMNE matrices) of the cells and particles in the aspirates using the hematology analyser ABX Pentra XL 80. Results: 112 patients (34.8%) had an infection according to the ICM criteria. When analysing the graphical LMNE matrices from synovia cell counting, four types could be differentiated: the type “wear particles” (I) in 28.3%, the type “infection” (II) in 24.8%, the “combined” type (III) in 15.5% and “indeterminate” type (IV) in 31.4%. There was a significant correlation between the graphical LMNE-types and the histological types of Morawietz and Krenn (p < 0.001 and Cramer test V value of 0.529). The addition of the LMNE-Matrix assessment increased the diagnostic value of the cell count and the cut-off value of the WBC count could be set lower by adding the LMNE-Matrix to the diagnostic procedure. Conclusion: The graphical representation of the cell count analysis of synovial aspirates is a new and helpful method for differentiating between real periprosthetic infections with an increased leukocyte count and false positive data resulting from wear particles. This new approach helps to increase the diagnostic value of cell count analysis in the diagnosis of PJI.

Advantage of menadione-catalyzed chemiluminescent assay for the determination of viable mammalian cell number

2012 ◽  
Vol 421 (2) ◽  
pp. 428-432 ◽  
Author(s):  
Shiro Yamashoji ◽  
Naoko Yoshikawa ◽  
Masayuki Kirihara ◽  
Toshihiro Tsuneyoshi
Keyword(s):  
Mammalian Cell ◽  
Cell Number

scholarly journals Blood Cell Count Using Deep Learning Semantic Segmentation

2019 ◽  
Author(s):  
Thanh Tran ◽  
Lam Binh Minh ◽  
Suk-Hwan Lee ◽  
Ki-Ryong Kwon
Keyword(s):  
Deep Learning ◽  
Blood Cell ◽  
Cell Count ◽  
Blood Cells ◽  
Blood Smear ◽  
White Blood Cells ◽  
Semantic Segmentation ◽  
Peripheral Blood Smear ◽  
Cell Counting ◽  
Blood Cell Count

Clinically, knowing the number of red blood cells (RBCs) and white blood cells (WBCs) helps doctors to make the better decision on accurate diagnosis of numerous diseases. The manual cell counting is a very time-consuming and expensive process, and it depends on the experience of specialists. Therefore, a completely automatic method supporting cell counting is a viable solution for clinical laboratories. This paper proposes a novel blood cell counting procedure to address this challenge. The proposed method adopts SegNet - a deep learning semantic segmentation to simultaneously segment RBCs and WBCs. The global accuracy of the segmentation of WBCs, RBCs, and the background of peripheral blood smear images obtains 89% when segment WBCs and RBCs from the background of blood smear images. Moreover, an effective solution to separate grouped or overlapping cells and cell count is presented using Euclidean distance transform, local maxima, and connected component labeling. The counting result of the proposed procedure achieves an accuracy of 93.3% for red blood cell count using dataset 1 and 97.38% for white blood cell count using dataset 2.

scholarly journals Cell-Tak coating may cause mis-normalization of Seahorse metabolic flux data

2021 ◽  
Author(s):  
Michal Šíma ◽  
Stanislava Martínková ◽  
Anežka Kafková ◽  
Jan Pala ◽  
Jan Trnka
Keyword(s):  
Metabolic Flux ◽  
Cell Number ◽  
Cell Counting ◽  
Fluorescent Detection ◽  
Data Normalization ◽  
Tissue Samples ◽  
Flux Data ◽  
Research Areas ◽  
Healthy State ◽  
A Cell

Metabolic flux investigations of cells and tissue samples are a rapidly advancing tool in diverse research areas. Reliable methods of data normalization are crucial for an adequate interpretation of results and to avoid a misinterpretation of experiments and incorrect conclusions. The most common methods for metabolic flux data normalization are to cell number, DNA and protein. Data normalization may be affected by a variety of factors, such as density, healthy state, adherence efficiency, or proportional seeding of cells. The mussel-derived adhesive Cell-Tak is often used to immobilize poorly adherent cells. Here we demonstrate that this coating may strongly affect the fluorescent detection of DNA leading to an incorrect and highly variable normalization of metabolic flux data. Protein assays are much less affected and cell counting can virtually completely remove the effect of the coating. Cell-Tak coating also affects cell shape in a cell line-specific manner and may change cellular metabolism. Based on these observations we recommend cell counting as a gold standard normalization method for Seahorse metabolic flux measurements with protein content as a reasonable alternative.

Mammalian cell growth regulation (reply)

Nature ◽  
1976 ◽  
Vol 263 (5577) ◽  
pp. 532-532
Author(s):  
HOLLEY
Keyword(s):  
Cell Growth ◽  
Mammalian Cell ◽  
Growth Regulation ◽  
Cell Growth Regulation ◽  
Mammalian Cell Growth
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