Countstar Mira FL

Fluorescence Cell Analyzer

Countstar Mira Fluorescence Cell Analyzer integrates AI intelligent algorithm and adopts patented fixed focus and optical zoom technology to realize the identification of cell characteristics. With trypan blue and AOPI staining methods, it helps to achieve accurate counting of all types of cells and supports GFP/RFP transfection experiments. The instrument is easy to operate, efficient in analysis and testing, saves valuable scientific research time, and helps scientific research laboratory personnel to achieve fast and efficient cell analysis result.

Core Advantages

• Compact Dimensions of the Countstar Mira FL to save valuable lab space
• Advance, AI-based Image Analysis Algorithms
• Intuitive Graphical User Interface
• Impressive Internal Data Storage Capacity
• Innovative 3x Optical Zoom Technology allows a unique look of cells and particles in the range from 1µm to 180µm.
• Multiple Applications: AO/PI cell density and viability analysis, GFP/RFP transfection efficiency studies, classical Trypan Blue based cell counts
• Economical Disposable Consumables are saving your time and costs

• Product details
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Product details

High Resolution Images: Provides Visual Evidence of Your Research

**8″, HD resolution touch screen, glove sensitive

3-fold selectable optical zoom: 5.0x, 6.6x, and 8.0x**

From left to right:

Chlorella spec. in bright field (green algae; 3-8 µm); 5.0x,
6.6x, and 8.0x magnification; Cell Counting BioApp

From left to right:

Cross-linked Agarose particles (20µm to 50µm): 5.0x, 6.6x,
and 8.0x magnification; Cell Counting BioApp

CHO cells, stained by AO/PI;

bright field image, with viability labels.
Green: detected algae cells, Blue: unspecific objects; Pink: aggregated cells

Same sample of CHOs as

above; Fluorescence view: Green indicators: viable cells;
Red: dead cells; Pink: aggregated cells

Simply Smart: AI-based Image Analysis Algorithms

To analyze and classify the huge variability of cell’s optical appearance in images, the Countstar® Mira FL uses the advance of Artificial Intelligence (AI) based image analysis algorithms. These self-learning image analysis tools are capable to analyze even complex image situations acquired by the Countstar® Mira FL. The implementation of cell shape analysis algorithms allows for a highly precise and reproducible analysis of the cell cycle status possible. Furthermore, the data can be used to interpret correlations between cell status, and the formation of cell clusters (aggregates, small spheroids), depending from the environmental
conditions.

Brilliant: 3- Fold Optical Zoom Technology

Our innovative 3-level, optical zooming technology (5.0x, 6.6x, and 8.0x ) enables the viewer to identify objects in a size range from 1 µm to 180µm. Simply select one of the pre-installed BioApps for the different classes of objects. The high-resolution of images allows for an in depth-look inside monitored cells. This extends the range of applications to a wide range of cellular objects in a single device.

Intuitive Software:AI-based BioApps for Multiple Cell Culture Applications

Convincing: Multiple Analysis Features

The advance, Artificial Intelligence (AI) based image analysis algorithms of the Countstar® Mira FL software deliver all essential cell viability parameters to evaluate the actual status of a cell culture. Along with cell density and viability, the cell size distribution, the cell shape factor, and any formation of cell clusters (aggregates) are analyzed as well. The relative fluorescence intensity delivers you data about the content of markers or fluorescent tagged products. The growth curve feature allows an immediate feedback about the kinetics of a running bio-process. Single cells can be classified even inside aggregates. Changes of the cell shape factor can be an important, early-stage indicator for changes in your cell culture. The data provided from the
Countstar® Mira FL enables better interpretation for users to optimize the dynamics of cell culture processes with mammalian, algae, and yeast cells.

Intuitive: Graphical User Interface with BioApps

The structured and inuitive graphical user interface of the Countstar® Mira FL was designed to make the daily challenged of the lab workflow more efficient. Customizable assay protocol templates can be adapted to varying characteristics of each cell line. The GUI allows a quick access to already acquired images and results stored in the internal database. An export of images, pdf reports, and csv files can be accomplished in a few simple steps.

Data menu: overview results single analysis

Size distribution histogram of a CHO culture, stained by AO/PI

Growth curve,comparing two cultivations to each other

M__ain menu showing the pre-installed BioApps

Versatile: Powerful BioApps for Various Cell Types

One of the unique features of the Countstar® Mira FL are the pre-installed BioApps. BioApps allow for a wide range of experiments to be performed on various organisms and delivers multiple data points to help determine cell culture status.

• Customization
  BioApps are individually designed to fit single cell type characteristics. Their configuration can be adapted, copied, and exported to other Countstar® Mira FL devices.
• *Analysis Speed
  *The plug and play versatility of the Countstar® Mira FL allows for cell culture data results in seconds. A simple click on the BioApp, naming your sample, and the system delivers results in seconds.
• Future-Oriented
  The large number of already pre-installed BioApps can be expanded at any time by the ability to download and install new customized assay protocols.
• *Service and Support
  *Benefit from our long experience in designing and customizing BioApps specifically to the
  varying characteristics of thousands of cell types.
• *Access to Results
  *All analysis data and images are pre-sorted by the BioApp names. The search for specific results can be facilitated by narrowing down the measurement period. Up to 50,000 analysis data sets are permanently and quickly accessible on the internal 128GB storage medium.

AO/PI Cell Density and Viability BioApps

The AO/PI dual dye fluorescence BioApp for determination of cell density and viability of eukaryotic cell cultures is based on the principle that both dyes, Acridine Orange (AO) and Propidium Iodide (PI), will intercalate between the nucleic acids of the chromosomes inside the nucleus. AO will permeate an intact nucleus membrane at any time to stain the DNA, emitting a green light at 525nm max., if excited at 480nm. PI is only capable to pass a disintegrating nucleus membrane of a dying (dead) cell. This dye will emit a red light with its amplitude at 615nm, if PI is excited at 525nm. The AO/PI staining kit utilizes the physico-chemical phenomenon of the FRET-effect (Foerster Resonance Energy Transfer). If samples of cells are excited at 480nm, the emitting light of the present AO dye in the nucleus at 525nm will be totally absorbed by the accumulated PI dye in the nuclei of dead (dying) cells, which then emit due to their excitation the red light at 615nm. The energy transfer ensures that no double light emission or overexposure of light will occur. Additionally, it also ensures that only nuclei containing cells will get stained and no akaryotes, like erythrocytes, will be included.

Dilution series of HEK293 cell sample,showng the high accuracy of Mira FL test results

High comparability in viability of different cell samples, tested in a Countstar® Mira FL and
_flow cytometer (B/C CytoFLEX) _

Result view screen of a CHO cell culture sample, stained by AO/PI, showing the bright field images with deactivated labelling. (5.0x magnification)

The same sample showing the fluorescence labelling results in an enlarged view – green circles**identify the viable cells, red circles identify the dead or dying cells**

GFP/RFP Transfection Efficiency BioApps

Using green or red fluorescence proteins (GFP / RFP) to bind them covalently as marker tags to the protein of interest, is a widely established technique to check the transfection efficiency of viral or plasmid vector-based genes. GFP/RFP are reliable tools to quantify the optimization of a protein manufacturing process, and is a fast, and highly precise method to determine the protein yield in many cGMP regulated manufacturing processes of APIs. This labelling technology has also become also an important tool in various gene therapeutic approaches to control the transfection efficiency of the genetical modifications. Compared to flow cytometry, the Countstar® Mira FL does not only deliver comparable results, but provides additionally images, proving the content of questioned cells. All images and data are permanently stored and offers a high level of traceability at any time. Additionally, the imaged based analysis is significantly faster, and needs only 20µL of sample liquid. The Countstar® Mira FL simplifies the analysis workflow, helping to streamline each optimization process.

Peak diagrams of samples 1+4(see below), showing GFP expressing HEK 293 cells, tested in a B/C CytoFLEx, showing a 27.8%, and a 95.9% transfection efficiency rate.
The test data of all 4 samples show an excellent level of
correlation between the two devices

Analysis results including image selection in fluorescence view of HEK293 cells, transfected by GFP

Trypan Blue Cell Density/Viability BioApps

The Trypan Blue based exclusion method is commonly used in many labs to monitor cell culture status and is the gold standard in determination of concentration and viabilty. Trypan Blue will permeate disintegrating outer membrane structures of dying animal cells, accumulating inside the cell, and staining the cytoplasma blue. Live cells will block the Trypan Blue dye from penetrating their membranes. The viable cells will still appear bright under a microscope. This optical appearance of living and dead cells can be precisely discriminated using the AI based image analysis algorithms of a Countstar® Mira FL. Beyond cell number of dead and viable cells as results, the image analysis of our analyzer delivers additional data about the cells in focus: Cell diameters, aggregate formation, and cell morphology.

The linear regression value of the total cell density proves the high accuracy and precision of the Countstar® Mira FL technology

Test results of the Mira FL show a higher consistency compared to manual counts in a hemocytometer. Undercounts in manual analysis are typical due to lower statistical base quantity.

Result and image overview of a human T-cell suspension,stained by Trypan Blue