Application Notes
Cellometer Publications
Posters
Product LiteratureApplication Notes
Enumeration and Viability of Nucleated Cells from Bone Marrow, Cord Blood, and Mobilized Peripheral Blood PDF
Isolation, Quantitation and Viability Analysis of Neonatal Cardiomyocytes using Cellometer PDF
Image-Based Analysis of Cell Cycle Using PI PDF
Concentration & Viability of PBMCs without Lysing PDF
Development of a Novel Method to Assess Primary Hepatocyte Concentration and Viability PDF
Yeast Concentration and Viability using Image-Based Fluorescence Analysis PDF
Automated Methods for Counting and Analyzing Stem Cell Samples PDF
Quantitative Measurement of GFP Transfection Rates in 60 Seconds - Cellometer Vision PDF
Direct Count of White Blood Cells from Peripheral Blood Sample without Lysing Red Blood Cells PDF
Rapid Detection of Apoptosis in Jurkat Cells with FITC Conjugated Annexin-V - Cellometer Vision PDF
Simple, Fast Determination of Viability by Staining Cells with Propidium Iodide - Cellometer Vision PDF
Automated Counting and Sizing Freshly Isolated Adipocytes with Minimal Sample Preparation PDF
Insect Cell Counting and Size Analysis Using Cellometer Auto T4 PDF
Cellometer Auto T4 Cell Counting Consistency PDF
Cellometer Tips for Counting Clumpy Cells PDF
Counting Yeast, Human Platelet, and Algae Using Cellometer Auto M10 PDF.
Cellattice: Monitoring Primary Neuron Growth and Development PDF
Cellometer Publications
Automated quantification of budding Saccharomyces cerevisiae using a novel image cytometry method
J Ind Microbiol Biotechnol. 2013 Apr 5, Laverty DJ, Kury AL, Kuksin D, Pirani A, Flanagan K, Chan LL.
The measurements of concentration, viability, and budding percentages of Saccharomyces cerevisiae are performed on a routine basis in the brewing and biofuel industries. Generation of these parameters is of great importance in a manufacturing setting, where they can aid in the estimation of product quality, quantity, and fermentation time of the manufacturing process. Specifically, budding percentages can be used to estimate the reproduction rate of yeast populations, which directly correlates with metabolism of polysaccharides and bioethanol production, and can be monitored to maximize production of bioethanol during fermentation. The traditional method involves manual counting using a hemacytometer, but this is time-consuming and prone to human error. In this study, we developed a novel automated method for the quantification of yeast budding percentages using Cellometer image cytometry. The automated method utilizes a dual-fluorescent nucleic acid dye to specifically stain live cells for imaging analysis of unique morphological characteristics of budding yeast. In addition, cell cycle analysis is performed as an alternative method for budding analysis. We were able to show comparable yeast budding percentages between manual and automated counting, as well as cell cycle analysis. The automated image cytometry method is used to analyze and characterize corn mash samples directly from fermenters during standard fermentation. Since concentration, viability, and budding percentages can be obtained simultaneously, the automated method can be integrated into the fermentation quality assurance protocol, which may improve the quality and efficiency of beer and bioethanol production processes. Read More
Accurate measurement of peripheral blood mononuclear cell concentration using image cytometry to eliminate RBC-induced counting error.
J Immunol Methods. 2013 Feb 28, Chan LL, Laverty DJ, Smith T, Nejad P, Hei H, Gandhi R, Kuksin D, Qiu J.
Peripheral blood mononuclear cells (PBMCs) have been widely researched in the fields of immunology, infectious disease, oncology, transplantation, hematological malignancy, and vaccine development. Specifically, in immunology research, PBMCs have been utilized to monitor concentration, viability, proliferation, and cytokine production from immune cells, which are critical for both clinical trials and biomedical research. The viability and concentration of isolated PBMCs are traditionally measured by manual counting with trypan blue (TB) using a hemacytometer. One of the common issues of PBMC isolation is red blood cell (RBC) contamination. The RBC contamination can be dependent on the donor sample and/or technical skill level of the operator. RBC contamination in a PBMC sample can introduce error to the measured concentration, which can pass down to future experimental assays performed on these cells. To resolve this issue, RBC lysing protocol can be used to eliminate potential error caused by RBC contamination. In the recent years, a rapid fluorescence-based image cytometry system has been utilized for bright-field and fluorescence imaging analysis of cellular characteristics (Nexcelom Bioscience LLC, Lawrence, MA). The Cellometer image cytometry system has demonstrated the capability of automated concentration and viability detection in disposable counting chambers of unpurified mouse splenocytes and PBMCs stained with acridine orange (AO) and propidium iodide (PI) under fluorescence detection. In this work, we demonstrate the ability of Cellometer image cytometry system to accurately measure PBMC concentration, despite RBC contamination, by comparison of five different total PBMC counting methods: (1) manual counting of trypan blue-stained PBMCs in hemacytometer, (2) manual counting of PBMCs in bright-field images, (3) manual counting of acetic acid lysing of RBCs with TB-stained PBMCs, (4) automated counting of acetic acid lysing of RBCs with PI-stained PBMCs, and (5) AO/PI dual staining method. The results show comparable total PBMC counting among all five methods, which validate the AO/PI staining method for PBMC measurement in the image cytometry method. Read More
Rapid quantification of pathogenic fungi by Cellometer image-based cytometry
Journal of Microbiological Methods, September 2012, Berkes CA, Chan LL, Wilkinson A, Paradis B
The objective of this study was to develop an image-based cytometric methodology for the quantification of viable pathogenic yeasts, which can offer
increased sensitivity and efficiency when compared to the traditional colony forming unit (CFU) assay. Live/dead yeast quantification by flow cytometry
has been previously demonstrated, however, adoption of flow cytometric detection of pathogenic yeasts has been limited for a number of practical reasons
including its high cost and biosafety considerations. Our studies focus on detection of two human fungal pathogens: Histoplasma capsulatum and Candida
albicans. H. capsulatum colonizes alveolar macrophages by replicating within the macrophage phagosome. Here, we quantitatively assess the growth of H.
capsulatum yeasts within RAW 264.7 macrophages using acridine orange/propidium iodide staining in combination with Cellometer image-based cytometry;
this method faithfully recapitulates growth trends as measured by traditional CFU enumeration, but with significantly increased sensitivity. Additionally,
we directly assess infection of bone marrow-derived macrophages with a GFP-expressing strain of C. albicans. To demonstrate that image-based cytometry can
be used as a tool to assess the susceptibility of fungi to antifungal drugs, we perform dose response experiments with the antifungal drugs amphotericin B
and itraconazole and show that image-based cytometry allows rapid assessment of the kinetics of cytotoxicity induced by these antifungals. Our methodology
offers a rapid, accurate, and economical means for detection and quantification of important human fungal pathogens, either alone or in association with
host cells. Read More
Rapid Image-based Cytometry for Comparison of Fluorescent Viability Staining Methods
Journal of fluorescence, September 2012. Chan LL, Wilkinson AR, Paradis BD, Lai N.
The ability to accurately measure cell viability is important for any cell-based research. Traditionally,
viability measurements have been performed using trypan blue exclusion method on hemacytometer, which allowed researchers to visually
distinguish viable from nonviable cells. However, the trypan blue method is often limited to only cell lines or primary cells that have
been rigorously purified. In the recent years, small desktop image-based cell counters have been developed for rapid cell concentration and
viability measurement due to advances in imaging and optics technologies as well as novel fluorescent stains. In this work, we employed the
Cellometer image-based cytometer to demonstrate the ability to simplify viability detection compared to the current methods. We compared
various fluorescence viability detection methods using single- or dual-staining technique. Single-staining method using nucleic acid stains
including ethidium bromide, propidium iodide, 7AAD, DAPI, Sytox Green and Sytox Red, and enzymatic stains including CFDA and Calcein AM were
performed. All stains produced comparable results to trypan blue exclusion method for cell line samples. Dual-staining method using AO/PI, CFDA/PI,
Calcein AM/PI and Hoechst 33342/PI that enumerates viable and non-viable cells was tested on primary cell samples with high debris contents. This
method allowed exclusion of cellular debris and non-nucleated cells from analysis, which can eliminate the need to perform purification step during
sample preparation, and improves the efficiency of viability detection method. Overall, these image-based fluorescent cell counters can simplify
assay procedures as well as capture images for visual confirmation. Read More
A novel image-based cytometry method for autophagy detection in living cells
Autophagy, September 2012. Leo Li-Ying Chan, Dee Shen, Alisha R. Wilkinson, Wayne Patton, Ning Lai, Eric Chan, Dmitry Kuksin, Bo Lin and Jean Qiu
Autophagy is an important cellular catabolic process that plays a variety of important roles,
including maintenance of the amino acid pool during starvation, recycling of damaged proteins and organelles, and clearance
of intracellular microbes. Currently employed autophagy detection methods include fluorescence microscopy, biochemical
measurement, SDS-PAGE and western blotting, but they are time consuming, labor intensive, and require much experience for
accurate interpretation. More recently, development of novel fluorescent probes have allowed the investigation of autophagy
via standard flow cytometry. However, flow cytometers remain relatively expensive and require a considerable amount of
maintenance. Previously, image-based cytometry has been shown to perform automated fluorescence-based cellular analysis
comparable to flow cytometry. In this study, we developed a novel method using the Cellometer image-based cytometer in
combination with Cyto-ID® Green dye for autophagy detection in live cells. Read More
Novel image cytometric method for detection of physiological and metabolic changes in Saccharomyces cerevisiae
Society for Industrial Microbiology and Biotechnology, August 2012. Chan LL, Kury A, Wilkinson A, Berkes C, Pirani A.
The studying and monitoring of physiological and metabolic changes in Saccharomyces cerevisiae (S. cerevisiae)
has been a key research area for the brewing, baking, and biofuels industries, which rely on these economically important yeasts to produce
their products. Specifically for breweries, physiological and metabolic parameters such as viability, vitality, glycogen, neutral lipid, and
trehalose content can be measured to better understand the status of S. cerevisiae during fermentation. Traditionally, these physiological and
metabolic changes can be qualitatively observed using fluorescence microscopy or flow cytometry for quantitative fluorescence analysis of
fluorescently labeled cellular components associated with each parameter. However, both methods pose known challenges to the end-users.
Specifically, conventional fluorescent microscopes lack automation and fluorescence analysis capabilities to quantitatively analyze large
numbers of cells. Although flow cytometry is suitable for quantitative analysis of tens of thousands of fluorescently labeled cells, the
instruments require a considerable amount of maintenance, highly trained technicians, and the system is relatively expensive to both purchase
and maintain. In this work, we demonstrate the first use of Cellometer Vision for the kinetic detection and analysis of vitality, glycogen,
neutral lipid, and trehalose content of S. cerevisiae. This method provides an important research tool for large and small breweries to study
and monitor these physiological behaviors during production, which can improve fermentation conditions to produce consistent and higher-quality
products. Read More
A novel method for kinetic measurements of rare cell proliferation using Cellometer image-based cytometry
J Immunol Methods, March 2012. Chan LL, Zhong X, Pirani A, Lin B.
Cell proliferation is an important assay for pharmaceutical and biomedical research to
test the effects of a variety of treatments on cultured primary cells or cell lines. For immunological studies, the
ability to perform rapid cell proliferation analysis allows the identification of potential biological reagents for
inducing or inhibiting immune cell proliferation. Current cell proliferation analysis methods employ flow cytometry
for fluorescence detection of CFSE-labeled cells. However, conventional flow cytometers require a considerable amount
of cells per sample, which becomes an issue for kinetic measurements with rare cell population due to the lack of
samples for flow cytometric analyses at multiple time points during proliferation period. Here we report the development
of a novel cell proliferation kinetic detection method for low cell concentration samples using the new Cellometer
Vision system. Read More
A rapid detection method for apoptosis and necrosis measurement using the Cellometer imaging cytometry
Apoptosis, December 2011. Chan LL, Lai N, Wang E, Smith T, Yang X, Lin B.
Apoptosis and necrosis play an important role in various aspects of preclinical pharmaceutical
drug discovery and validation. The ability to quickly determine the cytotoxic effect of chemical compounds on cancer cells
allows researchers to efficiently identify potential drug candidates for further development in the pharmaceutical discovery
pipeline. Recently, a new imaging cytometry system has been developed by Nexcelom Bioscience LLC (Lawrence, MA, USA) for
fluorescence-based cell population analysis. Read More
Cellometer Vision as an alternative to flow cytometry for cell cycle analysis, mitochondrial potential, and immunophenotyping
Cytometry Part A, April 2011. Leo L. Chan, Xuemei Zhong, Jean Qiu, Peter Y. Li, Bo Lin
Cell phenotyping and cell cycle analysis are two commonly used assays in both clinical
diagnosis and biomedical research. Cell phenotyping by identifying different biomarkers is essential for the diagnosis
of hematologic malignancy, sub-classifying diseases, monitoring response to treatment, predicting prognosis, detecting
rare cell populations and residual malignant cells. Read More
Rapid detection of ABC transporter interaction: Potential utility in pharmacology
Journal of Pharmacological and Toxicological Methods, November 2010. Robey RW, Lin B, Qiu J, Chan LL, Bates SE
We have developed a method to characterize the function and inhibition of ABC transporters using
an automated cell counter with fluorescence detection capability. The assay was performed using stably-transfected HEK293 cells
expressing P-gp, MRP1, or ABCG2 and examining transport of fluorescent substrates in the presence or absence of known inhibitors
and compared to results obtained with a flow cytometer. Read more
Direct concentration and viability measurement of yeast in corn mash using a novel imaging cytometry method
Journal of Industrial Microbiology Biotechnology, October 2010. Chan LL, Lyettefi EJ, Pirani A, Smith T, Qiu J, Lin B.
Worldwide awareness of fossil-fuel depletion and global warming has been increasing over
the last 30 years. Numerous countries, including the USA and Brazil, have introduced large-scale
industrial fermentation facilities for bioethanol, biobutanol, or biodiesel production. Most of
these biofuel facilities perform fermentation using standard baker's yeasts that ferment sugar
present in corn mash, sugar cane, or other glucose media. In research and development in the biofuel
industry, selection of yeast strains (for higher ethanol tolerance) and fermentation conditions
(yeast concentration, temperature, pH, nutrients, etc.) can be studied to optimize fermentation performance.
Yeast viability measurement is needed to identify higher ethanol-tolerant yeast strains, which may prolong
the fermentation cycle and increase biofuel output. In addition, yeast concentration may be optimized to
improve fermentation performance. Therefore, it is important to develop a simple method for concentration
and viability measurement of fermenting yeast. In this work, we demonstrate an imaging cytometry method for
concentration and viability measurements of yeast in corn mash directly from operating fermenters. It employs
an [Cellometer] automated cell counter, a dilution buffer, and staining solution from Nexcelom Bioscience to perform
enumeration. The proposed method enables specific fluorescence detection of viable and nonviable yeasts,
which can generate precise results for concentration and viability of yeast in corn mash. This method can
provide an essential tool for research and development in the biofuel industry and may be incorporated into
manufacturing to monitor yeast concentration and viability efficiently during the fermentation process.
Read more
Posters
Accurate PBMC Concentration Measurement using Image Cytometry to Eliminate RBC-Induced Counting Error PDF
A Novel Image-Based Cytometry Method for Autophagy Detection in Living Cells PDF
Rapid Quantification of Pathogenic Fungi by Cellometer Image-Based Cytometry PDF
A Novel Imaging Cytometry Method for Quantitative Cell Viability Assay PDF
Comparison of fluorescence methods for determining yeast viability using a novel automated image-based cell counting and viability system PDF
Surface Marker Based Direct Cell Concentration Measurements Using a High Sensitivity Imaging Cytometry Method PDF
A Novel Method for Kinetic Measurements of Rare Immune Cell Proliferation using Cellometer Image-Based Cytometry PDF
A Rapid Alternative Method for Cell Cycle Analysis Using Cellometer Vision PDF
A Novel Imaging Cytometry Method for Immunophenotyping PDF
Apoptosis Analysis of Jurkat Cells using the Cellometer® Vision PDF
Concentration and Viability Measurement of Yeast in Corn Mash using the Cellometer® Vision PDF
Concentration and Viability Measurement of PBMC using the Cellometer® Vision PDF
Obtaining Consistent and Accurate Cell Counting Results with Cellometer Automatic Cell Counters PDF
Product Literature
Cellometer® Mini Product Flyer PDF
Cellometer® Mini Spec Sheet PDF
Cellometer® Auto 1000 Product Flyer PDF
Cellomete®r Auto 1000 Spec Sheet PDF
Cellometer® Auto T4 Product Flyer PDF
Cellometer® Auto Spec Sheet PDF
Cellometer® Auto 2000 Product Flyer PDF
Cellometer® Auto 2000 Spec Sheet PDF
Cellometer® Auto X4 for Yeast Product Flyer PDF
Cellometer® Auto X4 Spec Sheet PDF
Cellometer® Vision Product Flyer PDF
Cellometer® Vision CBA Product Flyer PDF
Cellometer® Vision CBA Spec Sheet PDF
Running Cellometer on an Intel Mac PDF
Cellometer® Vision Product Literature PDF
Cellometer Auto T4 Cell Counting Comparison to Hemacytometer PDF
Cellometer® Auto T4 Product Literature PDF
Disposable Hemacytometer Product Literature PDF
Disposable Hemacytometer Technical Data PDF