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Posts Tagged ‘White Paper’

Comparison of Trypan Blue and Fluorescence-Based Viability Detection Methods Via Morphological Observation

Leo Chan | May 20, 2015 | No Comments | Cellometer Application News

It’s White Paper Wednesday! Read our featured white paper: Comparison of Trypan Blue and Fluorescence-Based Viability Detection Methods Via Morphological Observation

Trypan Blue vs. AOPI Cell ViabilityDetermining cell viability is a vital component in many biological experiments that range from standard cell culture to the investigation of pharmacological agents on tumor cells. One of the earliest and most common methods for measuring cell viability is the trypan blue (TB) exclusion assay [1, 2]. Over the last two decades, there have been various publications on comparing TB exclusion and fluorescence-based cellular viability assays [1, 3-5]. Previous results have shown that in a time-course measurement, TB exclusion assays reported significantly higher viabilities when compared to fluorescence-based methods [4, 5].

Trypan Blue vs. AOPI Cell Viability
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Automated Method for Determination of Infectious Dose (TCID50) using Celigo Imaging Cytometer

Leo Chan | May 6, 2015 | No Comments | Celigo Application News

It’s White Paper Wednesday! Read our featured white paper: Automated Method for Determination of Infectious Dose (TCID50) using Celigo Imaging Cytometer

celigo-confluence-vs-direct-cell-countingNexcelom’s Celigo imaging cytometer has been applied to provide automated, rapid assessment of viral infectivity in a range of plate formats [4]. Using f-theta optics, Celigo provides high quality, whole-well images using bright field and/or fluorescent illumination. Automated segmentation and analysis provide quantitative and objective output of CPE based on characteristic changes to the host cell monolayer.

PBMC
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Characterization of Breast Cancer Drugs via Mammosphere Morphometric Analysis Using Celigo Imaging Cytometer

Leo Chan | April 22, 2015 | No Comments | Celigo Application News

It’s White Paper Wednesday! Read our featured white paper: Characterization of Breast Cancer Drugs via Mammosphere Morphometric Analysis Using Celigo Imaging Cytometer

Celigo-bright-field-images-of-mammospheresA panel of cytotoxic drugs, including doxorubicin and paclitaxel were used to study their effects on various breast cancer cell lines such as MDA-MD-436, MCF-7, SKBR3 and MDA-MB-231. Results show that the Colony Counting application can also be used to evaluate the clonogenicity and self-renewal of cancer stem/tumor-initiating cells by automatically analyzing mammosphere populations. The Colony Counting application on Celigo provides an efficient, reproducible and automated method for assessing the number, size, and morphology of cancer spheroids within multiwell plates.

PBMC
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Measuring Physiological and Metabolic Characteristics of Yeast Health for Beer Fermentation Samples using the Cellometer Vision

Leo Chan | April 8, 2015 | No Comments | Cellometer Application News

It’s White Paper Wednesday! Read our featured white paper: Measuring Physiological and Metabolic Characteristics of Yeast Health for Beer Fermentation Samples using the Cellometer Vision

Yeast health for beer fermentationCharacteristics such as the viability, vitality, glycogen, neutral lipid, and trehalose content of yeast samples must be measured to better understand changes in physiological and metabolic status during fermentation [4-9]. Monitoring changes of these parameters during fermentation can improve current production processes, and a subset of these parameters (glycogen, neutral lipid, and trehalose content) has been shown to play an important role in predicting yeast viability both during and after fermentation [10, 11].

Yeast health for beer fermentation
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Comparing Fluorescence-Based Viability Detection Method using the Cellometer Vision

Leo Chan | March 25, 2015 | No Comments | Cellometer Application News

It’s White Paper Wednesday! Read our featured white paper: Comparing Fluorescence-Based Viability Detection Method using the Cellometer Vision

Cell viabilityIn this work, Cellometer Vision was employed to demonstrate rapid fluorescence-based
viability measurements and the comparison of various fluorescent staining methods. First, fluorescent nucleic acid stains that examine membrane integrity were tested and validated by comparing against the standard trypan blue exclusion method. Similarly, fluorescent enzymatic stains that examine metabolic activities were tested and validated against the standard trypan blue exclusion method. Third, nucleic acid and enzymatic stains were compared by measuring viabilities of Jurkat cells incubated at different temperatures of water bath.

Cell viability
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Bright field and Fluorescent Image Analysis for Screening Applications using the Celigo Imaging Cytometer

Leo Chan | March 11, 2015 | No Comments | Celigo Application News

It’s White Paper Wednesday! Read our featured white paper: Bright field and Fluorescent Image Analysis for Screening Applications using the Celigo Imaging Cytometer

celigo-label-free-growth-curvesHere, we present whole-well imaging and label-free bright field cell counting applications that allow the Celigo user to generate growth curves over time and monitor cell counts and confluence at the individual well level. These bright field applications permit the screening of drugs altering cell proliferation and growth rate. We also highlight the capacity of the Celigo to image large objects making it suitable for analysis of large structures such as embryoid bodies, tumor spheres or small organisms. Finally, the analysis of complex samples by imaging is a challenge for high-content screening applications.

PBMC
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Detection of Multiplexed GFP Reporters in Primary Articular Chondrocyte Cultures Using Cellometer Vision Image Cytometer

Leo Chan | February 25, 2015 | No Comments | Cellometer Application News

It’s White Paper Wednesday! Read our featured white paper: Detection of Multiplexed GFP Reporters in Primary Articular Chondrocyte Cultures Using Cellometer Vision Image Cytometer

GFPIn this work, we have developed an image cytometry method for detecting and monitoring the cell expansion and differentiation of articular chondrocytes in primary culture. First, the feasibility of utilizing image cytometry for detection of fluorescent is shown by comparing measured fluorescent positive cell populations to flow cytometry. Next, articular chondrocyte cultures were established in multi-well plates from either single or Cyan/eGFP double reporter mouse lines and grown for 20 days to test the utility of the fluorescence-based image cytometry system.

GFP
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The Historical Development of the Hemacytometer

Leo Chan | February 11, 2015 | No Comments | Cellometer Application News

It’s White Paper Wednesday! Read our featured white paper:The Historical Development of the Hemacytometer

Cell counting with a hemacytometerThe hemacytometer has been an essential tool for hematologists, medical practitioners, and biologists for over a century. Depending on where it is being used, the word has multiple spellings such as hemacytometer, hemocytometer, haemacytometer, or haemocytometer, but for consistency purposes the word “hemacytometer” will be used in this review. The prefix “hema”, “hemo”, “haema”, or “haemo” means blood, while “cytometer” meant a device to measure cells. The device was initially used by medical practitioners to analyze patient blood samples, which was the initial spark that created the field of hematology. The hemacytometer has gone through a series of major development in the 1800s and early 1900s.

Cell counting with a hemacytometer
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Accurately Count PBMC and Measure Viability in Presence of Residual RBC

Leo Chan | October 1, 2014 | No Comments | Cellometer Application News

It’s White Paper Wednesday! This month’s featured white paper: Accurately Count PBMC and Measure Viability in Presence of Residual RBC

PBMCIn this work, we have developed an image cytometry method for detecting and monitoring the cell expansion and differentiation of articular chondrocytes in primary culture. First, the feasibility of utilizing image cytometry for detection of fluorescent is shown by comparing measured fluorescent positive cell populations to flow cytometry. Next, articular chondrocyte cultures were established in multi-well plates from either single or Cyan/eGFP double reporter mouse lines and grown for 20 days to test the utility of the fluorescence-based image cytometry system.

PBMC
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Cellometer Fluorescent Cell Counters for Mouse Samples

Dmitry Kuksin | July 17, 2014 | No Comments | Cellometer Application News

Obtain fast, accurate and consistent cell viability and concentration measurements of primary murine samples in <30 seconds using the Cellometer K2!

The use of mouse models is common across multiple fields of study and the samples acquired greatly vary in type and complexity, from splenocytes to tissue digests to tail vein blood.

  • Tissue debris, cell debris, and red-blood cell contamination can lead to inconsistent cell counts from sample to sample when performing manual counting.
  • Nexcelom’s Cellometer instruments perform fluorescent based analysis using nucleic acid dyes Acridine Orange/ Propidium Iodide (AO/PI).
  • In the dual color, AO/PI viability assay, only nucleated cells are stained and counted. Non-nucleated cells and cellular debris are not stained and therefore are not counted.

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Accurately Count PBMC and Measure Viability in Presence of Residual RBC

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Learn More:
Accurately Count Mouse Primary Samples in the Presence Cell Debris »
Mouse Spleenocytes

Only nucleated cells are detected in each sample

Viability and Concentration are automatically reported after cell counting is completed.

Measurements are based on the fluorescent detection of live (green) and dead (red) cells only.

Non-nucleated cells (shown in the red circle) are not stained and are not counted.


Learn More:
Accurately Count PBMC and Measure Viability in Presence of Residual RBC »
PBMCs Stained with AOPI

Because platelets and mature red blood cells do not have a nucleus, they are not stained by nuclear staining dyes.

With the dual-fluorescence method, there is NO counting error from RBCs, platelets, or debris. Lysis of RBCs is not necessary.